RNF185

UniProt ID: Q96GF1
Organism: Homo sapiens
Review Status: COMPLETE
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Gene Description

RNF185 is a small (192 aa) membrane-anchored RING-type E3 ubiquitin-protein ligase (EC 2.3.2.27) and a close paralog of RNF5/RMA1 in the RNF5/RNF185-like family. It has a cytoplasmic N-terminal C3HC4 RING domain (catalytic Cys-39; the RING domain is responsible for ligase activity) and two C-terminal transmembrane helices that anchor it in membranes. RNF185 is an ER-membrane-resident ERAD ubiquitin ligase: it mediates the cotranslational ubiquitination and proteasomal degradation of the misfolded membrane protein CFTR (including CFTR-deltaF508), and it functions partly redundantly with RNF5 as an E3 ligase module that is central to CFTR degradation. Its expression is induced by the unfolded protein response and ER stress, and it protects cells from ER stress-induced apoptosis; it preferentially partners with the ER-associated E2 enzymes UBE2J1 and UBE2J2. Beyond canonical ERAD, RNF185 has documented ligase-dependent regulatory roles that use distinct ubiquitin chain topologies. At the mitochondrial outer membrane it builds K63-linked chains on the Bcl-2 family protein BNIP1 to promote selective mitochondrial autophagy via the autophagy receptor p62, and it builds non-degradative K27-linked chains on the DNA sensor cGAS (CGAS) at Lys-173/Lys-384 to enhance its enzymatic activity and the cGAS-STING innate antiviral response. RNF185 is ubiquitously expressed; its core function is as an ER-membrane ERAD ubiquitin ligase.

Existing Annotations Review

GO Term Evidence Action Reason
GO:0061630 ubiquitin protein ligase activity
IBA
GO_REF:0000033 		ACCEPT
Summary: Phylogenetic assignment of RING-type ubiquitin ligase activity, the core molecular function of RNF185, conserved across the RNF5/RNF185 family.
Reason: Core molecular function; supported by multiple IDA/EXP studies (EC 2.3.2.27; RING C39A abolishes activity).
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
The RING-type zinc finger domain is responsible for E3 ubiquitin ligase activity
GO:0036503 ERAD pathway
IBA
GO_REF:0000033 		ACCEPT
Summary: Phylogenetic assignment of involvement in ER-associated degradation, the core biological process of RNF185.
Reason: Core biological process; directly supported (CFTR ERAD) and conserved in the family.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway
GO:0044390 ubiquitin-like protein conjugating enzyme binding
IBA
GO_REF:0000033 		KEEP AS NON CORE
Summary: RNF185 binds ubiquitin-conjugating (E2) enzymes, preferentially the ERAD E2s UBE2J1 and UBE2J2. Informative but ancillary to the ligase activity.
Reason: Accurately reflects E2 binding required for catalysis, but is a mechanistic subsidiary of the core ubiquitin ligase activity.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Preferentially associates with the E2 enzymes UBE2J1 and UBE2J2
GO:0004842 ubiquitin-protein transferase activity
IEA
GO_REF:0000117 		ACCEPT
Summary: ARBA electronic assignment of ubiquitin-protein transferase activity, a parent of the specific RING-type ubiquitin ligase activity.
Reason: Correct general molecular function; the specific GO:0061630 captures the core RING E3 ligase activity.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
EC=2.3.2.27
GO:0005737 cytoplasm
IEA
GO_REF:0000117 		KEEP AS NON CORE
Summary: ARBA electronic assignment of generic cytoplasmic localization; RNF185 has a cytoplasmic N-terminus but is a membrane protein.
Reason: Generic and imprecise; the core localization is the ER membrane. The cytoplasmic topological domain does not make the protein a soluble cytoplasmic protein.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Cytoplasmic
GO:0005741 mitochondrial outer membrane
IEA
GO_REF:0000044 		KEEP AS NON CORE
Summary: Electronic transfer of mitochondrial outer membrane localization, reflecting the BNIP1/mitochondrial-autophagy study. A real but secondary localization.
Reason: Supported by PMID:21931693 but represents a secondary, autophagy-related localization distinct from the core ER-membrane ERAD function.
Supporting Evidence:
PMID:21931693
The two C-terminal transmembrane domains of human RNF185 mediate its localization to mitochondrial outer membrane
GO:0005783 endoplasmic reticulum
IEA
GO_REF:0000002 		ACCEPT
Summary: InterPro-based electronic assignment of ER localization, consistent with RNF185's ER-membrane ERAD function.
Reason: Correct compartment; redundant with the more specific ER membrane annotations and IDA evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
IEA
GO_REF:0000044 		ACCEPT
Summary: Electronic transfer of ER membrane localization from the UniProt subcellular location; the core compartment for RNF185's ERAD function.
Reason: Correct core localization; supported experimentally (EXP, PMID:27485036).
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Endoplasmic reticulum membrane {ECO:0000269|PubMed:24019521, ECO:0000269|PubMed:27485036}
GO:0006511 ubiquitin-dependent protein catabolic process
IEA
GO_REF:0000002 		ACCEPT
Summary: InterPro-based electronic assignment of ubiquitin-dependent protein catabolism, a parent process consistent with RNF185's ERAD role.
Reason: Correct but generic; the specific GO:0036503 (ERAD pathway) better captures the core biological role.
Supporting Evidence:
PMID:27485036
unfolded proteins that accumulate in the ER are transported to the cytosol for ubiquitin-proteasome-mediated degradation
GO:0036503 ERAD pathway
IEA
GO_REF:0000117 		ACCEPT
Summary: ARBA machine-learning electronic assignment of the ERAD pathway, redundant with the experimentally supported core process.
Reason: Correct core biological process; redundant with IMP/IGI and IBA evidence.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway
GO:0044322 endoplasmic reticulum quality control compartment
IEA
GO_REF:0000108 		KEEP AS NON CORE
Summary: Inter-ontology logical inference placing RNF185 in the ER quality control compartment, a plausible localization derived from its ERAD role.
Reason: Plausible localization inferred from the ERAD/ERQC link, but not directly demonstrated; the core localization annotation is ER membrane.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0046872 metal ion binding
IEA
GO_REF:0000002 		KEEP AS NON CORE
Summary: InterPro-based assignment of metal (zinc) ion binding by the RING domain, a structural requirement for the ligase fold.
Reason: Accurate structural feature of the RING domain but subsidiary to the informative ubiquitin ligase activity; not a standalone core function.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
RING-type
GO:0061630 ubiquitin protein ligase activity
IEA
GO_REF:0000120 		ACCEPT
Summary: Combined automated electronic assignment of the core RING E3 ligase activity, consistent with EC 2.3.2.27 and experimental evidence.
Reason: Correct core molecular function; redundant with IDA/EXP evidence.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
EC=2.3.2.27
GO:1904380 endoplasmic reticulum mannose trimming
IEA
GO_REF:0000117 		REMOVE
Summary: ARBA machine-learning assignment of ER mannose trimming. RNF185 is a ubiquitin ligase and does not trim mannose; this is pathway-adjacency over-annotation.
Reason: RNF185 has no glycosidase/mannosidase activity; mannose trimming is performed by EDEM/ER mannosidases. This electronic inference is biologically incorrect.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
E3 ubiquitin-protein ligase that regulates selective
GO:0005515 protein binding
IPI
PMID:19549727
Analysis of the human E2 ubiquitin conjugating enzyme protei... 		KEEP AS NON CORE
Summary: E2 ubiquitin-conjugating enzyme interaction network capturing RNF185 binding to UBE2D/UBE2E enzymes. Bare protein binding is uninformative.
Reason: Real E2 interactions relevant to catalysis but bare protein binding is uninformative; E2 binding is captured by GO:0044390.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Q96GF1; P51668: UBE2D1
GO:0005515 protein binding
IPI
PMID:25416956
A proteome-scale map of the human interactome network. 		KEEP AS NON CORE
Summary: Proteome-scale interactome map capturing RNF185 partners including RNF5. Bare protein binding is uninformative.
Reason: High-throughput interactome; bare protein binding is uninformative.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Q96GF1; Q99942: RNF5
GO:0005515 protein binding
IPI
PMID:28514442
Architecture of the human interactome defines protein commun... 		KEEP AS NON CORE
Summary: Interactome-community study capturing RNF185 binding to TMBIM6/BI-1. Bare protein binding is uninformative.
Reason: High-throughput interactome; bare protein binding is uninformative.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Q96GF1; P55061: TMBIM6
GO:0005515 protein binding
IPI
PMID:31515488
Extensive disruption of protein interactions by genetic vari... 		KEEP AS NON CORE
Summary: Interaction captured in a study of variant-driven interactome disruption. Bare protein binding is uninformative.
Reason: High-throughput interaction; bare protein binding is uninformative.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Q96GF1; Q969T4: UBE2E3
GO:0005515 protein binding
IPI
PMID:32296183
A reference map of the human binary protein interactome. 		KEEP AS NON CORE
Summary: Binary interactome reference map capturing many RNF185 membrane-protein partners. Bare protein binding is uninformative.
Reason: High-throughput interactome; bare protein binding is uninformative.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Q96GF1; P55061: TMBIM6
GO:0005515 protein binding
IPI
PMID:33961781
Dual proteome-scale networks reveal cell-specific remodeling... 		KEEP AS NON CORE
Summary: Cell-specific interactome capturing RNF185 partners including RNF5. Bare protein binding is uninformative.
Reason: High-throughput interactome; bare protein binding is uninformative.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Q96GF1; Q99942: RNF5
GO:0055085 transmembrane transport
TAS
Reactome:R-HSA-382556 		REMOVE
Summary: Reactome pathway-level annotation (ABC-family transport). RNF185 is a ubiquitin ligase, not a transporter; this reflects CFTR/ABC pathway-context bleed-through.
Reason: RNF185 does not mediate transmembrane transport; it is an E3 ligase acting on transporter substrates such as CFTR. The transport annotation is a pathway-adjacency artifact.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway
GO:0016567 protein ubiquitination
IEA
GO_REF:0000041 		KEEP AS NON CORE
Summary: UniPathway-derived general protein ubiquitination process, a parent of the specific ERAD ubiquitination RNF185 performs.
Reason: Correct but generic; the specific ERAD pathway and K63/K27 ubiquitination annotations better capture the role.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
PATHWAY: Protein modification; protein ubiquitination.
GO:0061630 ubiquitin protein ligase activity
TAS
Reactome:R-HSA-8866546 		ACCEPT
Summary: Reactome curation of RNF185 (with RNF5) ubiquitinating misfolded CFTR; captures the core ligase activity.
Reason: Correct core molecular function in the CFTR ERAD reaction.
Supporting Evidence:
PMID:24019521
identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
GO:0061630 ubiquitin protein ligase activity
TAS
Reactome:R-HSA-8867288 		ACCEPT
Summary: Reactome curation of the ERAD E3 ligase ubiquitinating an unfolded glycoprotein substrate; captures the core ligase activity.
Reason: Correct core molecular function.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
The RING-type zinc finger domain is responsible for E3 ubiquitin ligase activity
GO:0005741 mitochondrial outer membrane
EXP
PMID:21931693
RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates... 		KEEP AS NON CORE
Summary: Experimental localization to the mitochondrial outer membrane, where RNF185 ubiquitinates BNIP1 during mitochondrial autophagy. A real but secondary localization.
Reason: Directly supported (PMID:21931693) but a secondary compartment for the autophagy role, not the core ER ERAD function.
Supporting Evidence:
PMID:21931693
The two C-terminal transmembrane domains of human RNF185 mediate its localization to mitochondrial outer membrane
GO:0005789 endoplasmic reticulum membrane
EXP
PMID:27485036
Genome-wide identification and gene expression profiling of ... 		ACCEPT
Summary: Experimental evidence that RNF185 localizes to the ER membrane; the core compartment for its ERAD ligase activity.
Reason: Core localization with direct experimental support.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0061630 ubiquitin protein ligase activity
EXP
PMID:21931693
RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates... 		ACCEPT
Summary: Experimental demonstration of RNF185 ubiquitin ligase activity (K63-linked polyubiquitination of BNIP1). Core molecular function.
Reason: Core molecular function with direct experimental support.
Supporting Evidence:
PMID:21931693
human RNF185 is a mitochondrial ubiquitin E3 ligase that regulates selective mitochondrial autophagy in cultured cells
GO:0006511 ubiquitin-dependent protein catabolic process
IMP
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		ACCEPT
Summary: Mutant-phenotype evidence that RNF185 drives ubiquitin-dependent degradation of CFTR. Consistent with the core ERAD/proteasomal degradation role.
Reason: Supported by RNF185 depletion stabilizing CFTR; consistent with the core ERAD function.
Supporting Evidence:
PMID:24019521
its silencing stabilizes CFTR proteins
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		ACCEPT
Summary: Direct evidence of RNF185 RING-dependent ubiquitin ligase activity controlling CFTR stability. Core molecular function.
Reason: Core molecular function with direct experimental (IDA) support.
Supporting Evidence:
PMID:24019521
RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner
GO:0045089 positive regulation of innate immune response
IDA
PMID:28273161
The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated... 		KEEP AS NON CORE
Summary: Direct evidence that RNF185 positively regulates the cGAS-mediated innate immune response. A real but secondary, immunity-specific role.
Reason: Well supported (PMID:28273161) but a secondary moonlighting role distinct from the core ER ERAD function.
Supporting Evidence:
PMID:28273161
RNF185 specifically catalyzed the K27-linked poly-ubiquitination of cGAS, which promoted its enzymatic activity
GO:1904380 endoplasmic reticulum mannose trimming
TAS
Reactome:R-HSA-901032 		REMOVE
Summary: Reactome ERQC pathway annotation. RNF185 does not perform mannose trimming; this is pathway-adjacency over-annotation.
Reason: RNF185 is an E3 ligase with no mannosidase activity; ER mannose trimming is carried out by ER mannosidases/EDEMs. The annotation conflates pathway membership with direct activity.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
E3 ubiquitin-protein ligase that regulates selective
GO:0044314 protein K27-linked ubiquitination
IDA
PMID:28273161
The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated... 		KEEP AS NON CORE
Summary: Direct evidence that RNF185 builds non-degradative K27-linked polyubiquitin chains on cGAS, enhancing its activity. A secondary, immunity-related activity.
Reason: Directly demonstrated K27-linked ubiquitination, but in the context of the secondary cGAS-STING immune role rather than the core ERAD function. K27-linked chain building by RNF185 is corroborated on additional substrates (EBOV GP1,2, PMID:36224200; TUFM, PMID:38084826).
Supporting Evidence:
PMID:28273161
RNF185 specifically catalyzed the K27-linked poly-ubiquitination of cGAS
PMID:36224200
PMID:38084826
GO:0051607 defense response to virus
IDA
PMID:28273161
The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated... 		KEEP AS NON CORE
Summary: Direct evidence that RNF185 contributes to antiviral defense via the cGAS-STING pathway. A real but secondary, immunity-specific role.
Reason: Well supported but a secondary moonlighting role distinct from the core ER ERAD function.
Supporting Evidence:
PMID:28273161
the ER ubiquitin ligase RNF185 interacted with cGAS during HSV-1 infection
GO:0060340 positive regulation of type I interferon-mediated signaling pathway
IDA
PMID:28273161
The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated... 		KEEP AS NON CORE
Summary: Direct evidence that RNF185 positively regulates type I interferon signaling via cGAS. A real but secondary, immunity-specific role.
Reason: Well supported but a secondary moonlighting role distinct from the core ER ERAD function.
Supporting Evidence:
PMID:28273161
Ectopic-expression or knockdown of RNF185 respectively enhanced or impaired the IRF3-responsive gene expression
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:28273161
The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated... 		ACCEPT
Summary: Direct evidence of RNF185 ubiquitin ligase activity (K27-linked ubiquitination of cGAS; RING-dependent). Core molecular function.
Reason: Core molecular function with direct experimental support; C39A/C79A abolishes activity.
Supporting Evidence:
PMID:28273161
RNF185 specifically catalyzed the K27-linked poly-ubiquitination of cGAS
GO:0044877 protein-containing complex binding
IPI
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		KEEP AS NON CORE
Summary: RNF185 binds a protein complex (ERAD machinery) in the CFTR degradation study. More informative than bare protein binding but still ancillary.
Reason: Reflects association with the ERAD E3 module/machinery; supportive of, but subsidiary to, the core ligase and ERAD-process annotations.
Supporting Evidence:
PMID:24019521
identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
GO:0005783 endoplasmic reticulum
IDA
PMID:27485036
Genome-wide identification and gene expression profiling of ... 		ACCEPT
Summary: Direct evidence for ER localization, consistent with RNF185's ER-membrane ERAD function.
Reason: IDA-supported ER localization agrees with the documented ER-membrane site of action.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0043130 ubiquitin binding
IDA
PMID:27485036
Genome-wide identification and gene expression profiling of ... 		ACCEPT
Summary: Direct evidence that RNF185 binds ubiquitin, consistent with its in vitro autoubiquitination/ligase activity.
Reason: Supported by the in vitro autoubiquitination assays; reflects ubiquitin handling integral to ligase function.
Supporting Evidence:
PMID:27485036
in vitro autoubiquitination activity
GO:0051865 protein autoubiquitination
IDA
PMID:27485036
Genome-wide identification and gene expression profiling of ... 		KEEP AS NON CORE
Summary: RNF185 is autoubiquitinated in vitro, a common property of RING E3 ligases reflecting their catalytic activity.
Reason: Supported by in vitro autoubiquitination assays but a secondary property of the ligase, not its core substrate-directed function.
Supporting Evidence:
PMID:27485036
in vitro autoubiquitination activity
GO:0061630 ubiquitin protein ligase activity
IDA
PMID:27485036
Genome-wide identification and gene expression profiling of ... 		ACCEPT
Summary: Direct evidence of RNF185 E3 activity, with E3-activity-dependent resistance to ER stress. Core molecular function.
Reason: Core molecular function with direct experimental support; RING C39S decreases activity.
Supporting Evidence:
PMID:27485036
RNF185, but not CGRRF1 and RNF19B, exhibited significant resistance to ER stressor in an E3 activity-dependent manner
GO:1904294 positive regulation of ERAD pathway
IMP
PMID:27485036
Genome-wide identification and gene expression profiling of ... 		ACCEPT
Summary: Mutant-phenotype evidence that RNF185 positively regulates ERAD, protecting cells from ER stress in an E3-activity-dependent manner. Core biological process.
Reason: Core biological process; RNF185 is an ER-stress-induced ERAD E3 that confers ER stress resistance.
Supporting Evidence:
PMID:27485036
RNF185, but not CGRRF1 and RNF19B, exhibited significant resistance to ER stressor in an E3 activity-dependent manner
GO:0005515 protein binding
IPI
PMID:21931693
RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates... 		KEEP AS NON CORE
Summary: Interaction with BNIP1, a functionally important RNF185 substrate at the mitochondrial outer membrane. Bare protein binding is uninformative.
Reason: Records the real RNF185-BNIP1 interaction but bare protein binding is uninformative.
Supporting Evidence:
PMID:21931693
We further identified the Bcl-2 family protein BNIP1 as one of the substrates for RNF185
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8866542 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization within CFTR ERAD reactions. Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8866546 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (RNF5/RNF185 ubiquitinate misfolded CFTR). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8866551 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (CFTR binds ERAD machinery). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8866854 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (CFTR F508del translocation). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8866856 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (RNF5/RNF185 ubiquitinate CFTR F508del). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8866857 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (CFTR F508del binds ERAD machinery). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-8867288 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization in an ERAD ubiquitination reaction. Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-9931264 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (CD274/PD-L1 ERAD transport). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-9931298 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (ubiquitination of CD274 by ERAD complex). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005789 endoplasmic reticulum membrane
TAS
Reactome:R-HSA-9931313 		ACCEPT
Summary: Reactome curation of RNF185 ER membrane localization (p-CD274 binds ERAD complex). Core compartment.
Reason: Correct core localization; redundant with experimental evidence.
Supporting Evidence:
PMID:27485036
All were partially localised to the ER
GO:0005515 protein binding
IPI
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		KEEP AS NON CORE
Summary: Interaction captured in the RNF185/RNF5 CFTR ERAD study. Bare protein binding is uninformative.
Reason: Real interaction within the ERAD module but bare protein binding is uninformative.
Supporting Evidence:
PMID:24019521
identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
GO:0005783 endoplasmic reticulum
IDA
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		ACCEPT
Summary: Direct evidence for ER localization in the CFTR ERAD study. Core compartment.
Reason: IDA-supported ER localization agrees with the documented ER-membrane ERAD function.
Supporting Evidence:
PMID:24019521
RNF185 is a RING domain-containing polypeptide homologous to RNF5
GO:0005789 endoplasmic reticulum membrane
NAS
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		ACCEPT
Summary: Author-stated ER membrane localization in the CFTR ERAD study. Core compartment.
Reason: Consistent with experimentally supported ER-membrane localization.
Supporting Evidence:
PMID:24019521
RNF185 is a RING domain-containing polypeptide homologous to RNF5
GO:0036503 ERAD pathway
IGI
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		ACCEPT
Summary: Genetic-interaction evidence (with RNF5) that RNF185 drives ERAD of CFTR; co-depletion blocks CFTR-deltaF508 degradation. Core biological process.
Reason: Core biological process with direct experimental support from the RNF5/RNF185 co-depletion experiments.
Supporting Evidence:
PMID:24019521
simultaneous depletion of RNF5 and RNF185 profoundly blocks CFTRΔF508 degradation not only during translation but also after synthesis is complete
GO:0044390 ubiquitin-like protein conjugating enzyme binding
IPI
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		KEEP AS NON CORE
Summary: RNF185 binds the ERAD E2 enzymes UBE2J1/UBE2J2, demonstrated in the CFTR ERAD study. Informative but ancillary to ligase activity.
Reason: Reflects E2 (UBE2J1/UBE2J2) binding required for catalysis, but is a mechanistic subsidiary of the core ligase activity.
Supporting Evidence:
file:human/RNF185/RNF185-uniprot.txt
Preferentially associates with the E2 enzymes UBE2J1 and UBE2J2
GO:0051865 protein autoubiquitination
IDA
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associa... 		KEEP AS NON CORE
Summary: RNF185 autoubiquitinates, a common RING E3 property reflecting its catalytic activity.
Reason: Supported but a secondary property of the ligase, not its core substrate-directed function.
Supporting Evidence:
PMID:24019521
RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner

Core Functions

RING-type E3 ubiquitin ligase anchored in the ER membrane that mediates cotranslational ubiquitination of misfolded membrane-protein clients (notably CFTR and CFTR-deltaF508) for proteasomal degradation via the ERAD pathway, acting partly redundantly with RNF5 and preferentially using the E2 enzymes UBE2J1/UBE2J2.

Molecular Function:
ubiquitin protein ligase activity
Directly Involved In:
ERAD pathway
Cellular Locations:
endoplasmic reticulum membrane
Supporting Evidence:
  • PMID:24019521
    RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner
  • PMID:24019521
    identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation

ER-stress/UPR-induced ER-membrane ubiquitin ligase that positively regulates ERAD and confers resistance to ER stress-induced apoptosis in an E3-activity-dependent manner.

Molecular Function:
ubiquitin protein ligase activity
Directly Involved In:
positive regulation of ERAD pathway
Cellular Locations:
endoplasmic reticulum membrane
Supporting Evidence:
  • PMID:27485036
    RNF185, but not CGRRF1 and RNF19B, exhibited significant resistance to ER stressor in an E3 activity-dependent manner

References

GO_REF:0000002
Gene Ontology annotation through association of InterPro records with GO terms
GO_REF:0000033
Annotation inferences using phylogenetic trees
GO_REF:0000041
Gene Ontology annotation based on UniPathway vocabulary mapping
GO_REF:0000044
Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping
GO_REF:0000108
Automatic assignment of GO terms using logical inference, based on inter-ontology links
GO_REF:0000117
Electronic Gene Ontology annotations created by ARBA machine learning models
GO_REF:0000120
Combined Automated Annotation using Multiple IEA Methods
PMID:19549727
Analysis of the human E2 ubiquitin conjugating enzyme protein interaction network.
PMID:21931693
RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates autophagy through interaction with BNIP1.
  • RNF185 is a mitochondrial outer membrane E3 ligase that builds K63-linked polyubiquitin chains on BNIP1 to promote selective mitochondrial autophagy via recruitment of the autophagy receptor p62.
PMID:24019521
RNF185 is a novel E3 ligase of endoplasmic reticulum-associated degradation (ERAD) that targets cystic fibrosis transmembrane conductance regulator (CFTR).
  • RNF185 is an ER-membrane RING E3 that controls cotranslational ubiquitination and proteasomal degradation of CFTR/CFTR-deltaF508 in a RING- and proteasome-dependent manner; RNF185 and RNF5 form a partly redundant E3 module central to CFTR degradation, preferentially using E2 enzymes UBE2J1/UBE2J2.
PMID:25416956
A proteome-scale map of the human interactome network.
PMID:27485036
Genome-wide identification and gene expression profiling of ubiquitin ligases for endoplasmic reticulum protein degradation.
  • RNF185 is an ER-stress/UPR-induced ER-localized RING E3 with in vitro autoubiquitination activity that confers ER stress resistance in an E3-activity-dependent manner; the RING domain (Cys-39) is responsible for ligase activity.
PMID:28273161
The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated innate immune response.
  • ER-resident RNF185 interacts with cGAS during HSV-1 infection and catalyzes non-degradative K27-linked polyubiquitination of cGAS, enhancing its enzymatic activity and the cGAS-STING type I interferon antiviral response; RING residues Cys-39/Cys-79 are required.
PMID:28514442
Architecture of the human interactome defines protein communities and disease networks.
PMID:31515488
Extensive disruption of protein interactions by genetic variants across the allele frequency spectrum in human populations.
PMID:32296183
A reference map of the human binary protein interactome.
PMID:33961781
Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.
Reactome:R-HSA-382556
ABC-family proteins mediated transport
Reactome:R-HSA-8866542
VCP-catalyzed ATP hydrolysis promotes the translocation of misfolded CFTR into the cytosol
Reactome:R-HSA-8866546
RNF5 and RNF185 ubiquitinate misfolded CFTR
Reactome:R-HSA-8866551
CFTR binds components of the ERAD machinery for ubiquitination and degradation
Reactome:R-HSA-8866854
VCP-catalyzed ATP hydrolysis promotes the translocation of CFTR F508del into the cytosol
Reactome:R-HSA-8866856
RNF5 and RNF185 ubiquitinate CFTR F508del
Reactome:R-HSA-8866857
CFTR F508del binds components of the ERAD machinery for ubiquitination and degradation
Reactome:R-HSA-8867288
OS9:SEL1:ERAD E3 ligase:DERL2 ubiquitinates unfolded protein:(GlcNAc)2 (Man)9-5
Reactome:R-HSA-901032
ER Quality Control Compartment (ERQC)
Reactome:R-HSA-9931264
Active transport of ubiquitinated CD274 from ER to cytosol
Reactome:R-HSA-9931298
Ubiquitination of CD274 by ERAD complex
Reactome:R-HSA-9931313
p-S195-CD274 binds ERAD complex
PMID:36224200
RNF185 regulates proteostasis in Ebolavirus infection by crosstalk between the calnexin cycle, ERAD, and reticulophagy.
  • The ER ubiquitin ligase RNF185 polyubiquitinates misfolded Ebolavirus glycoprotein GP on Lys-673 via K27-linked chains, recruiting SQSTM1/p62 to divert GP to ATG3/ATG5-dependent reticulophagy/ERLAD (lysosomal) rather than proteasomal degradation.
PMID:38084826
Senecavirus A induces mitophagy to promote self-replication through direct interaction of 2C protein with K27-linked ubiquitinated TUFM catalyzed by RNF185.
  • RNF185 catalyzes K27-linked polyubiquitination of the mitochondrial translation elongation factor TUFM (via RNF185 transmembrane domain 1), enabling SQSTM1/p62 recognition and mitophagy that is exploited by Senecavirus A to promote replication.
PMID:39353943
Tapasin assembly surveillance by the RNF185/Membralin ubiquitin ligase complex regulates MHC-I surface expression.
  • RNF185 forms an ERAD complex with Membralin (MBRL/TMEM259) that recognizes unassembled Tapasin (a peptide-loading-complex component) and targets it for ubiquitin-dependent degradation; loss of RNF185/Membralin raises Tapasin steady-state levels and increases surface MHC-I, defining an ERAD-based control of antigen presentation.

Suggested Questions for Experts

Q: How is RNF185 partitioned between the ER membrane (ERAD/CFTR) and the mitochondrial outer membrane (BNIP1/mitophagy), and what governs this dual localization and substrate choice?

Q: To what extent are the RNF5 and RNF185 ERAD substrate repertoires overlapping versus distinct, and is their redundancy substrate-specific?

Suggested Experiments

Experiment: Reconstitute CFTR-deltaF508 ubiquitination in vitro with purified RNF185, RNF5, UBE2J1/UBE2J2 and Derlin cofactors to compare paralog activity and map chain linkages, and perform RNF5/RNF185 single and double knockouts with quantitative ubiquitinome profiling to define overlapping vs distinct ERAD substrates.

Experiment: Use proximity labeling (BioID/TurboID) of ER-anchored vs mitochondria-anchored RNF185 to map compartment-specific interactomes and test which substrates (CFTR, BNIP1, cGAS) depend on each localization.

Deep Research

Falcon

(RNF185-deep-research-falcon.md)
Research Report: Human RNF185 (UniProt Q96GF1) — Functional Annotation Falcon Edison Scientific Literature 36 citations 2 artifacts 2026-06-12T04:59:37.810786
Edison artifact artifact-00 (text/markdown)
## Context ID: pqac-00000035 The requested cycloheximide (CHX) chase experiments and their corresponding quantifications are shown in Figure 3b (parental vs. RN (image/png)

The research report should be a detailed narrative explaining the function, biological processes, and localization of the gene product. Citations should be given for all claims.

You should prioritize authoritative reviews and primary scientific literature when conducting research. You can supplement
this with annotations you find in gene/protein databases, but these can be outdated or inaccurate.

We are specifically interested in the primary function of the gene - for enzymes, what reaction is catalyzed, and what is the substrate specificity? For transporters, what is the substrate? For structural proteins or adapters, what is the broader structural role? For signaling molecules, what is the role in the pathway.

We are interested in where in or outside the cell the gene product carries out its function.

We are also interested in the signaling or biochemical pathways in which the gene functions. We are less interested in broad pleiotropic effects, except where these elucidate the precise role.

Include evidence where possible. We are interested in both experimental evidence as well as inference from structure, evolution, or bioinformatic analysis. Precise studies should be prioritized over high-throughput, where available.

Research Report: Human RNF185 (UniProt Q96GF1) — Functional Annotation

1) Target identity verification (critical disambiguation)

RNF185 in this report refers specifically to human RNF185 encoding E3 ubiquitin-protein ligase RNF185 (UniProt Q96GF1), a RING-type E3 ligase with C3HC4 RING catalytic domain and C-terminal transmembrane helices that anchor it to intracellular membranes. Primary literature explicitly studying “human RNF185” as a mitochondrial/ER membrane E3 ligase (e.g., Tang et al., 2011; Khouri et al., 2013) matches this identity and domain architecture, supporting that the reviewed studies map to the correct target (tang2011rnf185anovel pages 1-2, khouri2013rnf185isa pages 5-7).

2) Key concepts and definitions (current understanding)

2.1 RNF185 is a RING-type E3 ubiquitin ligase (EC 2.3.2.27)

RING E3 ligases catalyze transfer of ubiquitin from an E2~Ub conjugate to substrate lysine(s) and can build distinct ubiquitin chain linkages that encode different outcomes (e.g., proteasomal degradation vs selective autophagy cargo tagging). RNF185 has been experimentally demonstrated to require an intact RING domain for its ubiquitin ligase activity in cell and in vitro assays (khouri2013rnf185isa pages 5-7).

2.2 RNF185 is a membrane-embedded E3 that can function at the ER and mitochondria

RNF185 is unusual in that strong evidence supports two biologically relevant localizations:
- Mitochondrial outer membrane (MOM): RNF185 contains two C-terminal transmembrane domains sufficient/necessary for MOM targeting; topology mapping places its RING domain in the cytosol, consistent with ubiquitinating cytosolic-facing substrates on mitochondria (tang2011rnf185anovel pages 1-2, tang2011rnf185anovel pages 2-3).
- Endoplasmic reticulum (ER): independent biochemical work shows RNF185 is predominantly ER-localized and functions in ER-associated degradation (ERAD) with ERAD components and ER-resident E2s (khouri2013rnf185isa pages 5-7).

2.3 Non-canonical ubiquitin linkages: K27 and K63 in RNF185 biology

Multiple RNF185-dependent processes involve non-canonical linkages:
- K63-linked chains: associated with signaling and selective autophagy cargo tagging; RNF185 polyubiquitinates BNIP1 predominantly through K63-based linkages, enabling recruitment of autophagy receptor SQSTM1/p62 (tang2011rnf185anovel pages 1-2).
- K27-linked chains: increasingly recognized in immune and autophagy regulation; RNF185 catalyzes K27-linked ubiquitination of viral and mitochondrial substrates that are subsequently recognized by SQSTM1/p62 to engage lysosomal pathways (zhang2022rnf185regulatesproteostasis pages 1-2, chen2024senecavirusainduces pages 20-21).

3) Molecular function, domains, and enzymology (substrate specificity)

3.1 Catalytic requirements and E2 usage

Khouri et al. provide direct enzymology evidence: purified GST-RNF185 shows strong auto-ubiquitination with UbcH5c, weaker with UbcH6, and none with UbcH7 in vitro. RING mutations (C39/C42) or deletion of the RING domain abolish activity, demonstrating RING-dependence (khouri2013rnf185isa pages 5-7). In cells, RNF185 associates with ERAD machinery (Derlin-1/Erlin2) and preferentially co-precipitates ER membrane E2s UBE2J1 (Ubc6e) and UBE2J2 (khouri2013rnf185isa pages 5-7).

Recent CRISPR genetic interaction data in a CFTR-F508del ERAD context suggest an additional E2 relationship: UBE2D3 appears genetically consistent with acting as an E2 for both RNF5 and RNF185 in that pathway (riepe2024smallmoleculecorrectorsdivert pages 7-8).

3.2 Substrate specificity (what RNF185 ubiquitinates)

RNF185 does not appear to act as a broad, indiscriminate ERAD ligase; rather, available data support select substrate modules with distinct outcomes:

A) CFTR (including ΔF508/F508del mutant) — proteasomal ERAD
RNF185 ubiquitinates and promotes proteasome-dependent degradation of CFTR and CFTR-F508del, acting co-translationally and post-translationally, with functional redundancy/cooperation with RNF5 (khouri2013rnf185isa pages 1-2, khouri2013rnf185isa pages 10-11, khouri2013rnf185isa pages 11-13). Quantitatively, RNF185 overexpression reduced CFTR-F508 half-life from 44 to 29 minutes, and RNF185 knockdown stabilized labeled CFTR by ≥2-fold in pulse-label assays (khouri2013rnf185isa pages 10-11). Combined RNF5+RNF185 depletion produced stronger stabilization of CFTR-F508 (reported ~4.5-fold steady-state increase) than either alone (khouri2013rnf185isa pages 11-13).

B) BNIP1 — mitophagy cargo tagging (K63-linked)
Tang et al. identify the Bcl-2 family protein BNIP1 as an RNF185 substrate at mitochondria. BNIP1 is polyubiquitinated by RNF185 through K63-based linkages, recruiting SQSTM1/p62, which bridges ubiquitin to LC3 and promotes selective mitochondrial autophagy (mitophagy) (tang2011rnf185anovel pages 1-2).

C) Tapasin (TPSN) — ERAD “assembly surveillance” controlling MHC-I presentation
Van de Weijer et al. (2024) show tapasin (TPSN), an essential peptide-loading complex (PLC) component, is a substrate of an ERAD complex containing RNF185 and Membralin (MBRL/TMEM259). RNF185/MBRL preferentially recognizes unassembled tapasin, ubiquitinates it (requiring lysines in its cytosolic tail), and promotes turnover via ERAD (weijer2024tapasinassemblysurveillance pages 2-3, weijer2024tapasinassemblysurveillance pages 6-7). TPSN is normally short-lived with reported ~4 hour half-life (weijer2024tapasinassemblysurveillance pages 2-3). Functionally, loss of RNF185/MBRL increases TPSN steady-state abundance and increases surface peptide-loaded MHC-I in professional antigen-presenting cells (weijer2024tapasinassemblysurveillance pages 8-9).

D) Viral glycoprotein (EBOV GP1,2) — K27-linked ubiquitin and reticulophagy/ERLAD
Zhang et al. (2022) report that RNF185 polyubiquitinates Ebolavirus GP1,2 on Lys673 via K27-linkage, and this polyubiquitination drives recruitment to SQSTM1/p62 and lysosome-directed degradation in an ATG3/ATG5-dependent manner, indicating crosstalk between ERAD and ER-to-lysosome pathways (reticulophagy/ERLAD) (zhang2022rnf185regulatesproteostasis pages 1-2).

E) TUFM (mitochondrial translation elongation factor) — K27-linked ubiquitin and mitophagy
Chen et al. (2024) demonstrate RNF185 catalyzes K27-linked polyubiquitination of TUFM, enabling SQSTM1 recognition and mitophagy during Senecavirus A infection. Linkage specificity is supported by ubiquitin mutant mapping: only K27R ubiquitin significantly reduced TUFM polyubiquitination. Interaction mapping places the RNF185–TUFM interface on RNF185 TM1 (aa133–155) with TUFM interaction regions mainly in aa56–252 and a second site near ~344/345 (chen2024senecavirusainduces pages 20-21). RNF185 perturbation changes mitophagy markers (LC3-II, SQSTM1) and viral yield, linking this enzymatic activity to a functional phenotype (chen2024senecavirusainduces pages 19-20).

4) Subcellular localization and where RNF185 acts

4.1 Mitochondrial outer membrane RNF185

Tang et al. provide strong localization evidence using mitochondrial markers, biochemical fractionation, and proteinase K sensitivity of mitochondrial fractions, supporting RNF185 as a MOM protein with cytosolic-facing RING domain (tang2011rnf185anovel pages 2-3). This localization supports RNF185’s role in tagging mitochondrial proteins for mitophagy (BNIP1, TUFM-associated mitophagy) (tang2011rnf185anovel pages 1-2, chen2024senecavirusainduces pages 20-21).

4.2 ER membrane RNF185 in ERAD and ER proteostasis

Khouri et al. show RNF185 behaves as an ERAD E3 ligase: its ER targeting depends on its distal transmembrane segment, and it forms complexes with canonical ERAD factors (Derlin-1, Erlin2) and ER membrane E2s (UBE2J1/UBE2J2) (khouri2013rnf185isa pages 5-7). RNF185 is also transcriptionally induced under ER stress (tunicamycin) with a reported peak around 12 h, supporting a role in adaptive ER proteostasis (khouri2013rnf185isa pages 5-7).

4.3 RNF185/Membralin (MBRL) complex as an ERAD module

In the antigen presentation context, RNF185 operates as part of a multi-pass ERAD complex with MBRL, serving as an “assembly surveillance” mechanism for tapasin. Deletion of RNF185/MBRL markedly stabilizes TPSN in cycloheximide chase experiments (weijer2024tapasinassemblysurveillance pages 6-7). (weijer2024tapasinassemblysurveillance media 83ac0578, weijer2024tapasinassemblysurveillance media 08a702f3)

5) Pathways and biological processes

5.1 ER-associated degradation (ERAD) and proteostasis

RNF185 is implicated in ERAD at multiple levels:
- CFTR quality control: RNF185 is part of a functional module with RNF5 targeting CFTR and CFTR-F508del for proteasomal degradation, including co-translational ERAD (khouri2013rnf185isa pages 11-13, khouri2013rnf185isa pages 10-11).
- Tapasin assembly surveillance: RNF185/MBRL selectively limits availability of unassembled tapasin, tuning PLC formation and thereby MHC-I surface peptide presentation (weijer2024tapasinassemblysurveillance pages 1-2, weijer2024tapasinassemblysurveillance pages 8-9).
- Virus-perturbed ER proteostasis: RNF185 can couple ERAD-like recognition with autophagy/lysosomal routing through K27 ubiquitination and SQSTM1 recruitment (EBOV GP1,2) (zhang2022rnf185regulatesproteostasis pages 1-2).

5.2 Selective autophagy/mitophagy

RNF185 can mark mitochondrial or ER-associated cargo for selective autophagy:
- Mitophagy via BNIP1 (K63 ubiquitin–p62 axis) (tang2011rnf185anovel pages 1-2).
- Mitophagy via TUFM (K27 ubiquitin–p62 axis) in a viral infection model (chen2024senecavirusainduces pages 20-21).
- Reticulophagy/ERLAD via K27 ubiquitination of EBOV GP1,2 and SQSTM1 recruitment (zhang2022rnf185regulatesproteostasis pages 1-2).

6) Recent developments (prioritizing 2023–2024)

6.1 2024: RNF185/Membralin governs tapasin turnover and MHC-I surface expression

A major 2024 advance is the identification of tapasin as an RNF185/MBRL ERAD substrate controlling MHC-I presentation. Loss of RNF185/MBRL increased TPSN steady-state levels in multiple human cell models and increased surface peptide-loaded MHC-I in iPSC-derived macrophages (including after IFNγ stimulation), suggesting RNF185 is a potentially actionable lever for modulating immune surveillance (weijer2024tapasinassemblysurveillance pages 8-9, weijer2024tapasinassemblysurveillance pages 2-3). This work also provides mechanistic determinants of recognition, including requirements for tapasin tail lysines and a conserved intramembrane lysine affecting assembly and degradation decisions (weijer2024tapasinassemblysurveillance pages 6-7).

6.2 2024: RNF185-driven K27 ubiquitination of TUFM initiates mitophagy in viral infection

Chen et al. (2024) define a linkage-specific mechanism: RNF185 catalyzes K27-linked polyubiquitination of TUFM, which is then recognized by SQSTM1 to initiate mitophagy benefiting viral replication, and maps structural interaction domains (RNF185 TM1; TUFM segments) using truncations and docking (chen2024senecavirusainduces pages 20-21, chen2024senecavirusainduces pages 19-20).

6.3 2024: Genome-scale screens highlight RNF185 redundancy with RNF5 in CFTR-F508del ERAD

Riepe et al. (2024) provide a systems-level view: RNF185 is not necessarily rate-limiting alone, but becomes apparent in sensitized genetic backgrounds (RNF5 KO), indicating substantial pathway redundancy. Their work supports an E2 linkage with UBE2D3 and highlights that RNF5 perturbation enhances the effect of CFTR correctors (elexacaftor/tezacaftor) on CFTR-F508del half-life and maturation (riepe2024smallmoleculecorrectorsdivert pages 8-9, riepe2024smallmoleculecorrectorsdivert pages 7-8).

6.4 2023 review synthesis: positioning RNF185 among mitochondrial E3 ligases and cancer relevance

A 2023 review frames RNF185 as one of a small set of mitochondria-associated E3 ligases with emerging roles in autophagy, immune signaling, and cancer, while emphasizing that many mechanistic details and substrate scopes remain incomplete (gregorio2023roleofthe pages 3-5, gregorio2023roleofthe pages 9-10).

7) Current applications and real-world implementations

7.1 Cystic fibrosis (CFTR-F508del) — targetability of ubiquitination/ERAD

RNF185 has been proposed as part of a therapeutic target module (RNF5/RNF185) to stabilize CFTR variants (khouri2013rnf185isa pages 1-2). However, 2024 CRISPR work emphasizes that CFTR-F508del ERAD is robust and redundant, implying that single-target interventions may have limited effects and that combinatorial strategies (e.g., dual E3 targeting plus correctors) may be required (riepe2024smallmoleculecorrectorsdivert pages 8-9).

7.2 Immuno-oncology / antigen presentation modulation (MHC-I)

The RNF185/MBRL tapasin surveillance mechanism provides a plausible intervention axis: loss of RNF185/MBRL increases surface peptide-loaded MHC-I in professional antigen-presenting cells, implying that pharmacologic inhibition of RNF185/MBRL could potentially augment antigen presentation (weijer2024tapasinassemblysurveillance pages 8-9). This is not yet a clinical therapy, but represents a concrete, mechanistically defined lever with clear immunophenotypic readouts.

7.3 Antiviral strategy concepts

RNF185-dependent ubiquitination can be hijacked by viruses to promote their replication (e.g., SVA-induced mitophagy via TUFM) (chen2024senecavirusainduces pages 19-20) or to tune viral glycoprotein abundance/fitness via ER proteostasis routing (EBOV GP1,2) (zhang2022rnf185regulatesproteostasis pages 1-2). These studies suggest that RNF185 sits at a host-pathogen interface where inhibition could be antiviral in some contexts, though directionality may differ by virus (host defense vs viral exploitation).

8) Expert opinions / authoritative analysis

A key expert-level consensus emerging from recent review synthesis is that RNF185 is a multi-compartment, multi-output E3 ligase:
- It couples membrane localization (ER/MOM) to selective substrate modules (CFTR, tapasin, BNIP1, TUFM, viral GP1,2) with distinct fates (proteasome vs autophagy/lysosome) (gregorio2023roleofthe pages 12-13, gregorio2023roleofthe pages 3-5).
- It is best conceptualized as a context-dependent regulator of organelle proteostasis and immune-relevant processes, not as a single-pathway enzyme (gregorio2023roleofthe pages 9-10).

9) Statistics and quantitative data highlights

  • CFTR-F508del degradation kinetics: RNF185 overexpression reduces CFTR-F508 half-life 44 → 29 min; RNF185 knockdown increases labeled CFTR by ≥2-fold; combined RNF5+RNF185 depletion increases CFTR-F508 steady state by ~4.5-fold (Khouri 2013) (khouri2013rnf185isa pages 10-11, khouri2013rnf185isa pages 11-13).
  • Tapasin turnover: TPSN reported ~4 h half-life in control cells; RNF185/MBRL loss increases TPSN steady state and decreases TPSN ubiquitination (van de Weijer 2024) (weijer2024tapasinassemblysurveillance pages 2-3).
  • CRISPR pharmacogenetic interaction: RNF5 KO increases corrector effects on CFTR-F508del half-life and maturation by 52% and 46%, respectively; RNF185 is uncovered as redundant in sensitized screens (Riepe 2024) (riepe2024smallmoleculecorrectorsdivert pages 8-9).
  • TUFM ubiquitin linkage specificity: TUFM polyubiquitination is reduced specifically by K27R ubiquitin, indicating RNF185-catalyzed K27-linkage dependence (Chen 2024) (chen2024senecavirusainduces pages 20-21).

Evidence map (summary table)

Aspect Key finding Experimental evidence/assay Key quantitative/statistical notes Primary source (include first author year journal) and URL Citation ID
Molecular function RNF185 is a RING-dependent ERAD E3 ligase localized predominantly to the ER; it associates with ERAD factors and shows E2 preference for UbcH5c in vitro, with cell-based association to Ubc6e/UBE2J1 and UBE2J2 Purified GST-RNF185 in vitro auto-ubiquitination with E1/E2/ubiquitin; co-immunoprecipitation with Derlin-1, Erlin2, Ubc6e/UBE2J1, UBE2J2; RING mutants and TM truncation to test catalytic and localization requirements Robust auto-ubiquitination with UbcH5c, weaker with UbcH6, none with UbcH7; C39/C42 RING mutations or RING deletion abolished activity; distal TM truncation disrupted ER targeting; tunicamycin induced RNF185 transcripts peaking at ~12 h Khouri 2013, Journal of Biological Chemistry. https://doi.org/10.1074/jbc.m113.470500 (khouri2013rnf185isa pages 5-7)
Substrate / phenotype RNF185 targets CFTR and CFTR-ΔF508 for ubiquitin-proteasome degradation, acting with RNF5 as a major ERAD module for co- and post-translational control Pulse-labeling with [35S]Met/Cys, cycloheximide chase, proteasome inhibition (ALLN), overexpression and knockdown/silencing, anti-HA IP and quantification RNF185 overexpression reduced CFTR-F508 half-life from 44 to 29 min and reduced 35S-labeled CFTR signal by up to ~50%; RNF185 knockdown increased labeled CFTR at least ~2-fold; RNF5 knockdown ~3-fold stabilization, RNF185 knockdown ~2-fold, combined depletion ~4.5-fold stabilization of CFTR-F508 steady-state levels Khouri 2013, Journal of Biological Chemistry. https://doi.org/10.1074/jbc.m113.470500 (khouri2013rnf185isa pages 1-2, khouri2013rnf185isa pages 10-11, khouri2013rnf185isa pages 11-13)
Localization / pathway RNF185 is a mitochondrial outer membrane E3 ligase; its two C-terminal TM domains target it to mitochondria and it promotes selective mitochondrial autophagy/mitophagy via BNIP1 Confocal colocalization with MitoTracker and DsRed2-Mito; biochemical fractionation; proteinase K protection; GFP-LC3 puncta assays; LC3-I to LC3-II immunoblotting; siRNA knockdown; in vivo ubiquitination assays TM2 especially required for mitochondrial targeting; RNF185 overexpression increased LC3-II and GFP-LC3 puncta; RNF185 knockdown reduced basal LC3-II; BNIP1 was polyubiquitinated by RNF185 through K63-linked chains and recruited p62 Tang 2011, PLoS ONE. https://doi.org/10.1371/journal.pone.0024367 (tang2011rnf185anovel pages 1-2, tang2011rnf185anovel pages 2-3)
Substrate / pathway In ebolavirus infection, ER-associated RNF185 polyubiquitinates EBOV GP1,2 on Lys673 with K27-linked chains, diverting it to SQSTM1/p62-dependent reticulophagy/ERLAD rather than proteasomal degradation Infection-based proteostasis assays; ubiquitination mapping; dependence on SQSTM1/p62, ATG3, and ATG5; ER proteostasis framework linking calnexin cycle, ERAD, and ERLAD K27-linked polyubiquitination on GP1,2 Lys673; degradation proceeds via lysosome/autophagosome recruitment rather than proteasome; study concludes this increases viral fitness Zhang 2022, Nature Communications. https://doi.org/10.1038/s41467-022-33805-9 (zhang2022rnf185regulatesproteostasis pages 1-2)
Substrate / pathway / phenotype RNF185 forms an ERAD complex with Membralin (MBRL) that recognizes unassembled tapasin (TPSN), promotes its ubiquitination and degradation, and thereby limits MHC-I surface expression Quantitative proteomics (TMT-LC-MS/MS); IP-MS and co-IP; cycloheximide chase; ubiquitination assays; knockout/rescue in iPSCs, HEK293, THP-1, U2OS, and iPSC-derived macrophages; W6/32 flow cytometry for MHC-I TPSN half-life ~4 h in control cells; RNF185 or MBRL loss increased TPSN steady-state levels and reduced ubiquitinated TPSN despite higher total TPSN; TPSN tail 4K→A mutant poorly ubiquitinated; p<0.05 threshold in proteomics; n=5 parental and n=5 MBRL KO astrocyte samples in TMT experiment; IFNγ stimulation used at 100 ng/mL for 16 h; CHX quantification from n=3 experiments van de Weijer 2024, Nature Communications. https://doi.org/10.1038/s41467-024-52772-x (weijer2024tapasinassemblysurveillance pages 3-4, weijer2024tapasinassemblysurveillance pages 4-6, weijer2024tapasinassemblysurveillance pages 2-3, weijer2024tapasinassemblysurveillance pages 1-2, weijer2024tapasinassemblysurveillance pages 8-9, weijer2024tapasinassemblysurveillance pages 6-7, weijer2024tapasinassemblysurveillance pages 9-10)
Substrate / pathway During Senecavirus A infection, RNF185 catalyzes K27-linked polyubiquitination of mitochondrial TUFM, enabling SQSTM1 recognition and mitophagy that promotes viral replication Co-IP and ubiquitination assays with HA-Ub and Ub lysine mutants; deletion-mutant mapping; docking/residue mapping; siRNA knockdown and overexpression; western blot/autophagy-marker analysis; TCID50 viral titration Only K27R ubiquitin mutant significantly reduced TUFM polyubiquitination; RNF185 interacts with TUFM via RNF185 TM1 (aa133–155); TUFM interaction regions mapped mainly to aa56–252 and around aa344/345; RNF185 knockdown increased TUFM, SQSTM1, TIMM23, TOMM20 and decreased LC3-II; experiments performed in three independent biological replicates; viral yields decreased after RNF185 knockdown and increased after RNF185 overexpression Chen 2024, Autophagy. https://doi.org/10.1080/15548627.2023.2293442 (chen2024senecavirusainduces pages 19-20, chen2024senecavirusainduces pages 20-21, chen2024senecavirusainduces pages 21-23, chen2024senecavirusainduces pages 17-19, chen2024senecavirusainduces pages 1-2)
Pathway / phenotype CRISPR screens show RNF185 is a redundant ER-resident E3 for CFTR-F508del ERAD in parallel with RNF5; genetic data suggest UBE2D3 may act as an E2 for both RNF5 and RNF185 Genome-wide CRISPR/Cas9 stability screen; sensitized sublibrary screens in RNF5KO and RNF5/UBE2D3KO backgrounds; double-knockout analysis and degradation kinetics RNF5 was top E3 hit, but RNF5 KO only modestly reduced CFTR-F508del degradation; RNF185 emerged as a modest hit in sensitized screens; RNF185/UBE2D3KO kinetics were indistinguishable from UBE2D3KO, supporting same-pathway action; RNF5 disruption plus correctors increased mNG-F508del half-life and maturation by 52% and 46%, respectively; triple RNF185/RNF5/UBE2D3 knockout caused severe growth defect Riepe 2024, Molecular Biology of the Cell. https://doi.org/10.1091/mbc.e23-08-0336 (riepe2024smallmoleculecorrectorsdivert pages 8-9, riepe2024smallmoleculecorrectorsdivert pages 7-8, riepe2024smallmoleculecorrectorsdivert pages 1-2, riepe2024smallmoleculecorrectorsdivert pages 9-10)
Expert synthesis Recent expert review places RNF185 at the intersection of mitochondrial quality control, ERAD, autophagy/mitophagy, innate immunity, and cancer biology; known targets/processes include BNIP1, CFTR, cGAS, and RNF185/Membralin-dependent ER quality control Narrative synthesis of primary literature on mitochondrial E3 ligases and cancer/mitochondrial biology Review emphasizes context-dependent tumor-suppressive versus oncogenic roles, epigenetic/miRNA regulation, and major knowledge gaps in substrate scope and cooperation with other E3 ligases Di Gregorio 2023, International Journal of Molecular Sciences. https://doi.org/10.3390/ijms242417176 (gregorio2023roleofthe pages 7-9, gregorio2023roleofthe pages 3-5, gregorio2023roleofthe pages 12-13, gregorio2023roleofthe pages 5-7, gregorio2023roleofthe pages 9-10)

Table: This table summarizes experimentally supported and review-level evidence for human RNF185 (UniProt Q96GF1), covering its molecular function, localization, substrates, pathways, and phenotypic consequences. It is useful as a compact evidence map for functional annotation and literature-backed curation.

Figure evidence (example)

Cycloheximide chase panels in van de Weijer et al. (2024) directly visualize that tapasin (TPSN) degradation is blocked/stabilized in RNF185 KO and MBRL KO cells relative to parental controls, with associated quantification (weijer2024tapasinassemblysurveillance media 83ac0578, weijer2024tapasinassemblysurveillance media 08a702f3).

  1. Direct comparison of ER vs MOM pools of RNF185: existing work supports both localizations, but the degree of compartmental partitioning across cell types and stimuli remains incompletely quantified (khouri2013rnf185isa pages 5-7, tang2011rnf185anovel pages 2-3).
  2. Linkage and E2 mapping across substrates: strong linkage evidence exists for K63 (BNIP1) and K27 (EBOV GP1,2; TUFM), but systematic mapping of E2 partners and linkage types by substrate (and cell context) is still limited (tang2011rnf185anovel pages 1-2, chen2024senecavirusainduces pages 20-21, khouri2013rnf185isa pages 5-7).
  3. Therapeutic feasibility: CFTR-focused studies indicate redundancy in ERAD E3 usage, suggesting that RNF185 inhibition alone may not yield large functional rescue; however, immune modulation via tapasin surveillance may offer a distinct application domain with more direct phenotypic leverage (riepe2024smallmoleculecorrectorsdivert pages 8-9, weijer2024tapasinassemblysurveillance pages 8-9).

References

  1. (tang2011rnf185anovel pages 1-2): Fei Tang, Bin Wang, Na Li, Yanfang Wu, Junying Jia, Talin Suo, Quan Chen, Yong-Jun Liu, and Jie Tang. Rnf185, a novel mitochondrial ubiquitin e3 ligase, regulates autophagy through interaction with bnip1. PLoS ONE, 6:e24367, Sep 2011. URL: https://doi.org/10.1371/journal.pone.0024367, doi:10.1371/journal.pone.0024367. This article has 120 citations and is from a peer-reviewed journal.

  2. (khouri2013rnf185isa pages 5-7): Elma El Khouri, Gwenaëlle Le Pavec, Michel B. Toledano, and Agnès Delaunay-Moisan. Rnf185 is a novel e3 ligase of endoplasmic reticulum-associated degradation (erad) that targets cystic fibrosis transmembrane conductance regulator (cftr). Journal of Biological Chemistry, 288:31177-31191, Oct 2013. URL: https://doi.org/10.1074/jbc.m113.470500, doi:10.1074/jbc.m113.470500. This article has 124 citations and is from a domain leading peer-reviewed journal.

  3. (tang2011rnf185anovel pages 2-3): Fei Tang, Bin Wang, Na Li, Yanfang Wu, Junying Jia, Talin Suo, Quan Chen, Yong-Jun Liu, and Jie Tang. Rnf185, a novel mitochondrial ubiquitin e3 ligase, regulates autophagy through interaction with bnip1. PLoS ONE, 6:e24367, Sep 2011. URL: https://doi.org/10.1371/journal.pone.0024367, doi:10.1371/journal.pone.0024367. This article has 120 citations and is from a peer-reviewed journal.

  4. (zhang2022rnf185regulatesproteostasis pages 1-2): Jing Zhang, Bin Wang, Xiaoxiao Gao, Cheng Peng, Chao Shan, Silas F. Johnson, Richard C. Schwartz, and Yong-Hui Zheng. Rnf185 regulates proteostasis in ebolavirus infection by crosstalk between the calnexin cycle, erad, and reticulophagy. Nature Communications, Oct 2022. URL: https://doi.org/10.1038/s41467-022-33805-9, doi:10.1038/s41467-022-33805-9. This article has 38 citations and is from a highest quality peer-reviewed journal.

  5. (chen2024senecavirusainduces pages 20-21): Meirong Chen, Xin Zhang, Fanshu Kong, Peng Gao, Xinna Ge, Lei Zhou, Jun Han, Xin Guo, Yongning Zhang, and Hanchun Yang. Senecavirus a induces mitophagy to promote self-replication through direct interaction of 2c protein with k27-linked ubiquitinated tufm catalyzed by rnf185. Autophagy, 20:1286-1313, Jan 2024. URL: https://doi.org/10.1080/15548627.2023.2293442, doi:10.1080/15548627.2023.2293442. This article has 26 citations and is from a domain leading peer-reviewed journal.

  6. (riepe2024smallmoleculecorrectorsdivert pages 7-8): Celeste Riepe, Magda Wąchalska, Kirandeep K. Deol, Anais K. Amaya, Matthew H. Porteus, James A. Olzmann, and Ron R. Kopito. Small-molecule correctors divert cftr-f508del from erad by stabilizing sequential folding states. Molecular Biology of the Cell, Feb 2024. URL: https://doi.org/10.1091/mbc.e23-08-0336, doi:10.1091/mbc.e23-08-0336. This article has 10 citations and is from a domain leading peer-reviewed journal.

  7. (khouri2013rnf185isa pages 1-2): Elma El Khouri, Gwenaëlle Le Pavec, Michel B. Toledano, and Agnès Delaunay-Moisan. Rnf185 is a novel e3 ligase of endoplasmic reticulum-associated degradation (erad) that targets cystic fibrosis transmembrane conductance regulator (cftr). Journal of Biological Chemistry, 288:31177-31191, Oct 2013. URL: https://doi.org/10.1074/jbc.m113.470500, doi:10.1074/jbc.m113.470500. This article has 124 citations and is from a domain leading peer-reviewed journal.

  8. (khouri2013rnf185isa pages 10-11): Elma El Khouri, Gwenaëlle Le Pavec, Michel B. Toledano, and Agnès Delaunay-Moisan. Rnf185 is a novel e3 ligase of endoplasmic reticulum-associated degradation (erad) that targets cystic fibrosis transmembrane conductance regulator (cftr). Journal of Biological Chemistry, 288:31177-31191, Oct 2013. URL: https://doi.org/10.1074/jbc.m113.470500, doi:10.1074/jbc.m113.470500. This article has 124 citations and is from a domain leading peer-reviewed journal.

  9. (khouri2013rnf185isa pages 11-13): Elma El Khouri, Gwenaëlle Le Pavec, Michel B. Toledano, and Agnès Delaunay-Moisan. Rnf185 is a novel e3 ligase of endoplasmic reticulum-associated degradation (erad) that targets cystic fibrosis transmembrane conductance regulator (cftr). Journal of Biological Chemistry, 288:31177-31191, Oct 2013. URL: https://doi.org/10.1074/jbc.m113.470500, doi:10.1074/jbc.m113.470500. This article has 124 citations and is from a domain leading peer-reviewed journal.

  10. (weijer2024tapasinassemblysurveillance pages 2-3): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  11. (weijer2024tapasinassemblysurveillance pages 6-7): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  12. (weijer2024tapasinassemblysurveillance pages 8-9): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  13. (chen2024senecavirusainduces pages 19-20): Meirong Chen, Xin Zhang, Fanshu Kong, Peng Gao, Xinna Ge, Lei Zhou, Jun Han, Xin Guo, Yongning Zhang, and Hanchun Yang. Senecavirus a induces mitophagy to promote self-replication through direct interaction of 2c protein with k27-linked ubiquitinated tufm catalyzed by rnf185. Autophagy, 20:1286-1313, Jan 2024. URL: https://doi.org/10.1080/15548627.2023.2293442, doi:10.1080/15548627.2023.2293442. This article has 26 citations and is from a domain leading peer-reviewed journal.

  14. (weijer2024tapasinassemblysurveillance media 83ac0578): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  15. (weijer2024tapasinassemblysurveillance media 08a702f3): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  16. (weijer2024tapasinassemblysurveillance pages 1-2): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  17. (riepe2024smallmoleculecorrectorsdivert pages 8-9): Celeste Riepe, Magda Wąchalska, Kirandeep K. Deol, Anais K. Amaya, Matthew H. Porteus, James A. Olzmann, and Ron R. Kopito. Small-molecule correctors divert cftr-f508del from erad by stabilizing sequential folding states. Molecular Biology of the Cell, Feb 2024. URL: https://doi.org/10.1091/mbc.e23-08-0336, doi:10.1091/mbc.e23-08-0336. This article has 10 citations and is from a domain leading peer-reviewed journal.

  18. (gregorio2023roleofthe pages 3-5): Jacopo Di Gregorio, Martina Appignani, and Vincenzo Flati. Role of the mitochondrial e3 ubiquitin ligases as possible therapeutic targets in cancer therapy. International Journal of Molecular Sciences, 24:17176, Dec 2023. URL: https://doi.org/10.3390/ijms242417176, doi:10.3390/ijms242417176. This article has 15 citations.

  19. (gregorio2023roleofthe pages 9-10): Jacopo Di Gregorio, Martina Appignani, and Vincenzo Flati. Role of the mitochondrial e3 ubiquitin ligases as possible therapeutic targets in cancer therapy. International Journal of Molecular Sciences, 24:17176, Dec 2023. URL: https://doi.org/10.3390/ijms242417176, doi:10.3390/ijms242417176. This article has 15 citations.

  20. (gregorio2023roleofthe pages 12-13): Jacopo Di Gregorio, Martina Appignani, and Vincenzo Flati. Role of the mitochondrial e3 ubiquitin ligases as possible therapeutic targets in cancer therapy. International Journal of Molecular Sciences, 24:17176, Dec 2023. URL: https://doi.org/10.3390/ijms242417176, doi:10.3390/ijms242417176. This article has 15 citations.

  21. (weijer2024tapasinassemblysurveillance pages 3-4): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  22. (weijer2024tapasinassemblysurveillance pages 4-6): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  23. (weijer2024tapasinassemblysurveillance pages 9-10): Michael L. van de Weijer, Krishna Samanta, Nikita Sergejevs, LuLin Jiang, Maria Emilia Dueñas, Tiaan Heunis, Timothy Y. Huang, Randal J. Kaufman, Matthias Trost, Sumana Sanyal, Sally A. Cowley, and Pedro Carvalho. Tapasin assembly surveillance by the rnf185/membralin ubiquitin ligase complex regulates mhc-i surface expression. Nature Communications, Oct 2024. URL: https://doi.org/10.1038/s41467-024-52772-x, doi:10.1038/s41467-024-52772-x. This article has 10 citations and is from a highest quality peer-reviewed journal.

  24. (chen2024senecavirusainduces pages 21-23): Meirong Chen, Xin Zhang, Fanshu Kong, Peng Gao, Xinna Ge, Lei Zhou, Jun Han, Xin Guo, Yongning Zhang, and Hanchun Yang. Senecavirus a induces mitophagy to promote self-replication through direct interaction of 2c protein with k27-linked ubiquitinated tufm catalyzed by rnf185. Autophagy, 20:1286-1313, Jan 2024. URL: https://doi.org/10.1080/15548627.2023.2293442, doi:10.1080/15548627.2023.2293442. This article has 26 citations and is from a domain leading peer-reviewed journal.

  25. (chen2024senecavirusainduces pages 17-19): Meirong Chen, Xin Zhang, Fanshu Kong, Peng Gao, Xinna Ge, Lei Zhou, Jun Han, Xin Guo, Yongning Zhang, and Hanchun Yang. Senecavirus a induces mitophagy to promote self-replication through direct interaction of 2c protein with k27-linked ubiquitinated tufm catalyzed by rnf185. Autophagy, 20:1286-1313, Jan 2024. URL: https://doi.org/10.1080/15548627.2023.2293442, doi:10.1080/15548627.2023.2293442. This article has 26 citations and is from a domain leading peer-reviewed journal.

  26. (chen2024senecavirusainduces pages 1-2): Meirong Chen, Xin Zhang, Fanshu Kong, Peng Gao, Xinna Ge, Lei Zhou, Jun Han, Xin Guo, Yongning Zhang, and Hanchun Yang. Senecavirus a induces mitophagy to promote self-replication through direct interaction of 2c protein with k27-linked ubiquitinated tufm catalyzed by rnf185. Autophagy, 20:1286-1313, Jan 2024. URL: https://doi.org/10.1080/15548627.2023.2293442, doi:10.1080/15548627.2023.2293442. This article has 26 citations and is from a domain leading peer-reviewed journal.

  27. (riepe2024smallmoleculecorrectorsdivert pages 1-2): Celeste Riepe, Magda Wąchalska, Kirandeep K. Deol, Anais K. Amaya, Matthew H. Porteus, James A. Olzmann, and Ron R. Kopito. Small-molecule correctors divert cftr-f508del from erad by stabilizing sequential folding states. Molecular Biology of the Cell, Feb 2024. URL: https://doi.org/10.1091/mbc.e23-08-0336, doi:10.1091/mbc.e23-08-0336. This article has 10 citations and is from a domain leading peer-reviewed journal.

  28. (riepe2024smallmoleculecorrectorsdivert pages 9-10): Celeste Riepe, Magda Wąchalska, Kirandeep K. Deol, Anais K. Amaya, Matthew H. Porteus, James A. Olzmann, and Ron R. Kopito. Small-molecule correctors divert cftr-f508del from erad by stabilizing sequential folding states. Molecular Biology of the Cell, Feb 2024. URL: https://doi.org/10.1091/mbc.e23-08-0336, doi:10.1091/mbc.e23-08-0336. This article has 10 citations and is from a domain leading peer-reviewed journal.

  29. (gregorio2023roleofthe pages 7-9): Jacopo Di Gregorio, Martina Appignani, and Vincenzo Flati. Role of the mitochondrial e3 ubiquitin ligases as possible therapeutic targets in cancer therapy. International Journal of Molecular Sciences, 24:17176, Dec 2023. URL: https://doi.org/10.3390/ijms242417176, doi:10.3390/ijms242417176. This article has 15 citations.

  30. (gregorio2023roleofthe pages 5-7): Jacopo Di Gregorio, Martina Appignani, and Vincenzo Flati. Role of the mitochondrial e3 ubiquitin ligases as possible therapeutic targets in cancer therapy. International Journal of Molecular Sciences, 24:17176, Dec 2023. URL: https://doi.org/10.3390/ijms242417176, doi:10.3390/ijms242417176. This article has 15 citations.

Artifacts

Citations

  1. riepe2024smallmoleculecorrectorsdivert pages 7-8
  2. weijer2024tapasinassemblysurveillance pages 2-3
  3. weijer2024tapasinassemblysurveillance pages 8-9
  4. chen2024senecavirusainduces pages 20-21
  5. chen2024senecavirusainduces pages 19-20
  6. weijer2024tapasinassemblysurveillance pages 6-7
  7. riepe2024smallmoleculecorrectorsdivert pages 8-9
  8. gregorio2023roleofthe pages 9-10
  9. weijer2024tapasinassemblysurveillance pages 1-2
  10. gregorio2023roleofthe pages 3-5
  11. gregorio2023roleofthe pages 12-13
  12. weijer2024tapasinassemblysurveillance pages 3-4
  13. weijer2024tapasinassemblysurveillance pages 4-6
  14. weijer2024tapasinassemblysurveillance pages 9-10
  15. chen2024senecavirusainduces pages 21-23
  16. chen2024senecavirusainduces pages 17-19
  17. chen2024senecavirusainduces pages 1-2
  18. riepe2024smallmoleculecorrectorsdivert pages 1-2
  19. riepe2024smallmoleculecorrectorsdivert pages 9-10
  20. gregorio2023roleofthe pages 7-9
  21. gregorio2023roleofthe pages 5-7
  22. 35S
  23. https://doi.org/10.1074/jbc.m113.470500
  24. https://doi.org/10.1371/journal.pone.0024367
  25. https://doi.org/10.1038/s41467-022-33805-9
  26. https://doi.org/10.1038/s41467-024-52772-x
  27. https://doi.org/10.1080/15548627.2023.2293442
  28. https://doi.org/10.1091/mbc.e23-08-0336
  29. https://doi.org/10.3390/ijms242417176
  30. https://doi.org/10.1371/journal.pone.0024367,
  31. https://doi.org/10.1074/jbc.m113.470500,
  32. https://doi.org/10.1038/s41467-022-33805-9,
  33. https://doi.org/10.1080/15548627.2023.2293442,
  34. https://doi.org/10.1091/mbc.e23-08-0336,
  35. https://doi.org/10.1038/s41467-024-52772-x,
  36. https://doi.org/10.3390/ijms242417176,

📚 Additional Documentation

Notes

(RNF185-notes.md)

RNF185 review notes

UniProt: Q96GF1 (RN185_HUMAN), 192 aa, gene RNF185. HGNC:26783. Chromosome 22. Close paralog of RNF5/RMA1 (RNF5/RNF185-like family, IPR045103).

Core identity

RNF185 is a membrane-anchored RING-type E3 ubiquitin ligase (EC 2.3.2.27). C3HC4 RING domain (ZN_FING 39-80; catalytic Cys-39; C39A or C39A/C79A abolishes ligase activity) plus two C-terminal transmembrane helices (131-151, 172-192) with cytoplasmic N-terminus. The RING domain is responsible for E3 ligase activity.

[file:human/RNF185/RNF185-uniprot.txt "The RING-type zinc finger domain is responsible for E3 ubiquitin ligase activity"]
[file:human/RNF185/RNF185-uniprot.txt "MUTAGEN 39 ... C->A: Abolished E3 ubiquitin-protein ligase activity"]

Primary role: ERAD (CFTR), partly redundant with RNF5

RNF185 is an ER-membrane ERAD E3 ligase. It controls the cotranslational ubiquitination and degradation of CFTR/CFTRdeltaF508, in a RING- and proteasome-dependent manner, and forms an E3 ligase module with RNF5 that is central to CFTR degradation. Co-depletion of RNF5 + RNF185 profoundly blocks CFTRdeltaF508 degradation. RNF185 physically interacts with RNF5 (IntAct EBI-2340249/EBI-348482).

[file:human/RNF185/RNF185-uniprot.txt "Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway"]
PMID:24019521
PMID:27485036

RNF185 is upregulated by the UPR / ER stress (thapsigargin, tunicamycin) and protects cells from ER stress-induced apoptosis. It positively regulates ERAD (GO:1904294, IMP PMID:27485036).

[file:human/RNF185/RNF185-uniprot.txt "Protects cells from ER stress-induced apoptosis"]

E2 partners

Preferentially associates with the ERAD E2 enzymes UBE2J1 and UBE2J2; also binds UBE2D family in interactome screens.
[file:human/RNF185/RNF185-uniprot.txt "Preferentially associates with the E2 enzymes UBE2J1 and UBE2J2"]

Secondary / non-core roles

  • Mitochondrial autophagy: K63-linked polyubiquitination of BNIP1 at the mitochondrial outer membrane, stimulating LC3II accumulation and autophagolysosome formation; recruits p62. This is the original 2011 characterization ("mitochondrial E3 ligase").
    PMID:21931693
    PMID:21931693
  • Innate antiviral immunity: K27-linked polyubiquitination of cGAS (CGAS) at K173/K384, promoting cGAS enzymatic activity and cGAS-STING signaling / type I IFN. RNF185 is the first E3 ligase of cGAS.
    PMID:28273161
    [file:human/RNF185/RNF185-uniprot.txt "catalyzing 'Lys-27'-linked polyubiquitination of CGAS at 'Lys-173' and 'Lys-384', thereby promoting CGAS cyclic GMP-AMP synthase activity"]

Localization

UniProt records both Mitochondrion outer membrane (PMID:21931693) and ER membrane (PMID:24019521, PMID:27485036). The 2011 study reported mitochondrial outer membrane; later ERAD studies firmly place it at the ER membrane. ER membrane is the core compartment for its ERAD function; mitochondrial OM is a real but secondary localization tied to the BNIP1/autophagy role. Cytoplasm IEA (ARBA) is generic.

Annotation review decisions (summary)

  • ubiquitin protein ligase activity (GO:0061630): CORE, ACCEPT (multiple IDA/EXP; EC 2.3.2.27; RING C39A abolishes).
  • ERAD pathway (GO:0036503) / positive regulation of ERAD (GO:1904294): CORE, ACCEPT (IGI/IMP).
  • ER membrane (GO:0005789) / ER (GO:0005783): CORE location, ACCEPT.
  • ubiquitin-like protein conjugating enzyme binding (GO:0044390): KEEP_AS_NON_CORE (E2/UBE2J binding; ancillary).
  • ubiquitin binding (GO:0043130, IDA): ACCEPT non-core (reflects RING/ubiquitin handling).
  • protein autoubiquitination (GO:0051865): KEEP_AS_NON_CORE (RING property).
  • metal ion binding (GO:0046872) / zinc: KEEP_AS_NON_CORE (RING structural).
  • K27-linked ubiquitination (GO:0044314), defense response to virus (GO:0051607), positive regulation of innate immune response (GO:0045089), positive regulation of type I IFN signaling (GO:0060340): KEEP_AS_NON_CORE (cGAS immunity role, real but secondary).
  • mitochondrial outer membrane (GO:0005741): KEEP_AS_NON_CORE (BNIP1/autophagy role).
  • cytoplasm (GO:0005737, IEA ARBA): KEEP_AS_NON_CORE (generic).
  • protein binding (GO:0005515, many IPI): KEEP_AS_NON_CORE (uninformative).
  • protein-containing complex binding (GO:0044877): KEEP_AS_NON_CORE.
  • transmembrane transport (GO:0055085, Reactome TAS): REMOVE (RNF185 is not a transporter; CFTR/ABC pathway bleed-through).
  • ER mannose trimming (GO:1904380, IEA ARBA / Reactome TAS): REMOVE (no mannosidase activity; pathway-adjacency over-annotation).
  • ER quality control compartment (GO:0044322, IEA inter-ontology): KEEP_AS_NON_CORE.
  • ubiquitin-protein transferase activity (GO:0004842, IEA): ACCEPT (parent of ligase activity).
  • ubiquitin-dependent protein catabolic process (GO:0006511) / protein ubiquitination (GO:0016567): ACCEPT/KEEP_AS_NON_CORE (generic parents of ERAD).

Falcon deep-research findings (incorporated 2026-06)

Reviewed RNF185-deep-research-falcon.md (Edison/Falcon, 2026-06-12). Existing core ERAD/CFTR, RNF5 redundancy, BNIP1 K63 mitophagy, and cGAS K27/innate-immunity roles are already captured. Genuinely new RNF185-specific findings (all PubMed-verified, added to references):

  • NEW substrate Tapasin and the RNF185/Membralin (MBRL/TMEM259) ERAD complex: RNF185/Membralin recognizes unassembled Tapasin and degrades it via ERAD, controlling peptide-loading-complex formation and MHC-I surface expression; loss of the complex raises Tapasin and increases surface MHC-I. This is the strongest new finding and defines a named ER ligase complex plus a new physiological substrate. PMID:39353943 (Nat Commun 2024; DOI 10.1038/s41467-024-52772-x). Added as reference (relevance HIGH); full text not cached, so no GO annotation/supporting_text added. Candidate future annotations: protein-containing complex (RNF185/Membralin), and an ERAD/antigen-presentation MF/BP once full text is available.
  • NEW K27-linked substrate TUFM: RNF185 catalyzes K27-linked polyubiquitination of mitochondrial elongation factor TUFM (via RNF185 TM1, aa133-155), recruiting SQSTM1/p62 to drive mitophagy, exploited by Senecavirus A. PMID:38084826 (Autophagy 2024; DOI 10.1080/15548627.2023.2293442). Added as reference (relevance MEDIUM) and as a corroborating reference id on the existing K27-linked ubiquitination annotation (GO:0044314).
  • NEW K27-linked substrate EBOV GP1,2: RNF185 polyubiquitinates Ebolavirus glycoprotein GP on Lys-673 with K27 chains, diverting it to SQSTM1/p62-dependent reticulophagy/ERLAD (lysosomal) rather than the proteasome — crosstalk among calnexin cycle, ERAD and reticulophagy. PMID:36224200 (Nat Commun 2022; DOI 10.1038/s41467-022-33805-9). Added as reference (relevance MEDIUM) and as a corroborating reference id on GO:0044314.
  • Synthesis: across cGAS, EBOV-GP, and TUFM, RNF185 repeatedly builds non-degradative K27-linked chains read by SQSTM1/p62 to route cargo to autophagy/lysosome — reinforcing K27 + p62 as a recurring RNF185 signalling output distinct from its core K48/proteasomal ERAD (CFTR) activity.
  • Riepe et al. 2024 (MBoC, DOI 10.1091/mbc.e23-08-0336) genome-wide CRISPR screen: RNF185 is a redundant CFTR-F508del ERAD E3 uncovered only in RNF5-KO sensitized backgrounds, with UBE2D3 genetically acting as a shared E2; corroborates existing RNF5/RNF185 redundancy. Peripheral/not cached; not added as a reference.
  • Di Gregorio et al. 2023 review (IJMS, DOI 10.3390/ijms242417176) frames RNF185 among mitochondrial E3 ligases (autophagy, immune signalling, cancer); review-level context, not added.

Pn Notes

(RNF185-pn-notes.md)

RNF185 PN Consistency Notes

  • Generated: 2026-06-18
  • Project: PROTEOSTASIS
  • Scope: PN consistency rereview against local AIGR review and available deep-research artifacts
  • UniProt: Q96GF1
  • AIGR review status: COMPLETE
  • Review batch: proteostasis-batch-2026-06-11
  • Batch change status: added

Source Files Checked

Deep Research Files

AIGR Review Snapshot

  • Description: RNF185 is a small (192 aa) membrane-anchored RING-type E3 ubiquitin-protein ligase (EC 2.3.2.27) and a close paralog of RNF5/RMA1 in the RNF5/RNF185-like family. It has a cytoplasmic N-terminal C3HC4 RING domain (catalytic Cys-39; the RING domain is responsible for ligase activity) and two C-terminal transmembrane helices that anchor it in membranes. RNF185 is an ER-membrane-resident ERAD ubiquitin ligase: it mediates the cotranslational ubiquitination and proteasomal degradation of the misfolded membrane protein CFTR (including CFTR-deltaF508), and it functions partly redundantly with RNF5 as an E3 ligase module that is central to CFTR degradation. Its expression is induced by the unfolded protein response and ER stress, and it protects cells from ER stress-induced apoptosis; it preferentially partners with the ER-associated E2 enzymes UBE2J1 and UBE2J2. Beyond canonical ERAD, RNF185 has documented ligase-dependent regulatory roles that use distinct ubiquitin chain topologies. At the mitochondrial outer membrane it builds K63-linked chains on the Bcl-2 family protein BNIP1 to promote selective mitochondrial autophagy via the autophagy receptor p62, and it builds non-degradative K27-linked chains on the DNA sensor cGAS (CGAS) at Lys-173/Lys-384 to enhance its enzymatic activity and the cGAS-STING innate antiviral response. RNF185 is ubiquitously expressed; its core function is as an ER-membrane ERAD ubiquitin ligase.
  • Existing/core annotation action counts: ACCEPT: 32; KEEP_AS_NON_CORE: 23; REMOVE: 3

PN Consistency Summary

  • Consistency: Excellent. Deep research, review and PN concur: ER RING E3 (Cys-39), CFTR/ΔF508 ERAD redundant with RNF5; non-core BNIP1 K63 mitophagy (PMID:21931693), cGAS K27/antiviral (PMID:28273161), and the RNF185/Membralin/TMUB1-2 ER-membrane ligase complex. PN row 4 reference "32738194" verified via PubMed = van de Weijer et al. 2020 Mol Cell "Quality Control of ER Membrane Proteins by the RNF185/Membralin Ubiquitin Ligase Complex" (DOI) — the same complex the review cites via PMID:39353943 (Tapasin/Membralin). No contradictions.
  • PN story / NEW pressure: GO:0061630 + GO:0036503 already in GOA/review. GO:0000423 mitophagy (verified real) is new_to_goa — RNF185's BNIP1 role supports selective mitochondrial autophagy, a DEFENSIBLE ADD (review captures it only as BNIP1-substrate ligase activity + K63, no mitophagy BP term). GO:0000151 ubiquitin ligase complex (verified real, NOT in GOA) is well-grounded by the Membralin complex (PMID:32738194/39353943) — DEFENSIBLE ADD; review has GO:0044877 (complex binding) but no complex CC term. Conclude: catalytic/ERAD captured; GO:0000423 and GO:0000151 both defensible NEW.
  • Evidence alignment: Strong. PN mitophagy row PMID:21931693 = review BNIP1 annotation; PN membralin PMID:32738194 = review's Membralin/Tapasin literature (PMID:39353943, also van de Weijer). PN RING row "19489725 / rev" not in review (family review; low impact).
  • Verdict: Fully consistent; ligase MF + ERAD already captured, GO:0000423 mitophagy and GO:0000151 ubiquitin ligase complex both defensible NEW (non-core). No YAML change strictly required.

Full Consistency Review

  • UniProt: Q96GF1 · batch: proteostasis-batch-2026-06-11 · review status: COMPLETE (very thorough; ER RING E3, ERAD/CFTR core + BNIP1 mitophagy, cGAS/K27, Membralin/Tapasin)
  • PN placement: 4 rows — ER proteostasis|...|ERAD|...|ERAD-associated RING E3 ligase; ALP|Autophagy substrate selection|Autophagy receptor regulation|Mitophagy; UPS|...|RING|with transmembrane domain|ER, mitochondria; UPS|...|idiosyncratic RING complex|membralin complex|catalytic / RING, transmembrane. PN-node mapping: ERAD-RING subtype→GO:0061630 (in_goa); ERAD type/group→GO:0036503 (in_goa); Mitophagy type→GO:0000423 mitophagy (new_to_goa); RING group→GO:0061630 (in_goa); membralin-complex group→GO:0000151 ubiquitin ligase complex (new_to_goa). Projected: GO:0036503×2, GO:0061630×2 (in GOA), GO:0000423 (new), GO:0000151 (new).
  • Consistency: Excellent. Deep research, review and PN concur: ER RING E3 (Cys-39), CFTR/ΔF508 ERAD redundant with RNF5; non-core BNIP1 K63 mitophagy (PMID:21931693), cGAS K27/antiviral (PMID:28273161), and the RNF185/Membralin/TMUB1-2 ER-membrane ligase complex. PN row 4 reference "32738194" verified via PubMed = van de Weijer et al. 2020 Mol Cell "Quality Control of ER Membrane Proteins by the RNF185/Membralin Ubiquitin Ligase Complex" (DOI) — the same complex the review cites via PMID:39353943 (Tapasin/Membralin). No contradictions.
  • PN story / NEW pressure: GO:0061630 + GO:0036503 already in GOA/review. GO:0000423 mitophagy (verified real) is new_to_goa — RNF185's BNIP1 role supports selective mitochondrial autophagy, a DEFENSIBLE ADD (review captures it only as BNIP1-substrate ligase activity + K63, no mitophagy BP term). GO:0000151 ubiquitin ligase complex (verified real, NOT in GOA) is well-grounded by the Membralin complex (PMID:32738194/39353943) — DEFENSIBLE ADD; review has GO:0044877 (complex binding) but no complex CC term. Conclude: catalytic/ERAD captured; GO:0000423 and GO:0000151 both defensible NEW.
  • Mapping strategy: Correct and conservative. Membralin-complex group→GO:0000151 (complex membership, not catalytic activity to every subunit) is sound; consider the more specific child GO:0000835 ER ubiquitin ligase complex (verified in OLS) given the ER-membrane context. Mitophagy via receptor-regulation framing → GO:0000423 appropriate. No node-status change warranted.
  • Evidence alignment: Strong. PN mitophagy row PMID:21931693 = review BNIP1 annotation; PN membralin PMID:32738194 = review's Membralin/Tapasin literature (PMID:39353943, also van de Weijer). PN RING row "19489725 / rev" not in review (family review; low impact).
  • Verdict: Fully consistent; ligase MF + ERAD already captured, GO:0000423 mitophagy and GO:0000151 ubiquitin ligase complex both defensible NEW (non-core). No YAML change strictly required.
  • Recommended edits: [YAML] (optional) add GO:0000151 ubiquitin ligase complex (or child GO:0000835 ER ubiquitin ligase complex; part_of, PMID:32738194) and GO:0000423 mitophagy (involved_in, PMID:21931693) as non-core annotations to mirror the PN projections. [MAP] consider GO:0000835 ER ubiquitin ligase complex as a more precise target for the membralin-complex node.

PN Dossier Context

  • review_batch: proteostasis-batch-2026-06-11
  • review_yaml: genes/human/RNF185/RNF185-ai-review.yaml
  • PN workbook rows: 4

PN row 1: ER proteostasis | Organelle-specific protein degradation | ER associated degradation | Cytosolic handling of ERAD substrates | ERAD-associated RING E3 ligase

  • UniProt: Q96GF1
  • In branches: ER, ALP, UPS
  • PN-node mapping records (path + ancestors):
    • [subtype] ER proteostasis|Organelle-specific protein degradation|ER associated degradation|Cytosolic handling of ERAD substrates|ERAD-associated RING E3 ligase
      status=mapped scope=ok_for_propagation_to_go GO=[GO:0061630 ubiquitin protein ligase activity]
      rationale: This PN subtype denotes ERAD-associated RING E3 ligases. Ubiquitin protein ligase activity is the appropriate shared catalytic target.
    • [type] ER proteostasis|Organelle-specific protein degradation|ER associated degradation|Cytosolic handling of ERAD substrates
      status=mapped scope=ok_for_propagation_to_go GO=[GO:0036503 ERAD pathway]
      rationale: This PN type covers the cytosolic processing steps that receive ERAD substrates after retrotranslocation. These activities remain part of the ERAD pathway, but the source category is a specific mechanistic slice.
    • [group] ER proteostasis|Organelle-specific protein degradation|ER associated degradation
      status=mapped scope=exact GO=[GO:0036503 ERAD pathway]
      rationale: The PN group "ER associated degradation" is a direct lexical and biological match to the GO ERAD pathway term. The additional branch and class context disambiguates the source string from any broader degradation language.
    • [class] ER proteostasis|Organelle-specific protein degradation
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a broad PN category rather than a single GO class. The member genes span multiple activities, complexes, or contexts, so direct propagation from this node would overstate the shared biology.
    • [branch] ER proteostasis
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a top-level PN branch. This is a systems/taxonomy umbrella, not a direct GO assertion; narrower child curations carry any propagating GO mappings.

PN row 2: Autophagy-Lysosome Pathway | Autophagy substrate selection | Autophagy receptor regulation | Mitophagy

  • UniProt: Q96GF1
  • In branches: ER, ALP, UPS
  • Notes: Mitochondrial ubiquitin E3 ligase that regulates selective mitochondrial autophagy via interaction with BNIP1.
  • PN references (titles):
    • RNF185, a Novel Mitochondrial Ubiquitin E3 Ligase, Regulates Autophagy through Interaction with BNIP1 (plos.org)
  • PN-node mapping records (path + ancestors):
    • [type] Autophagy-Lysosome Pathway|Autophagy substrate selection|Autophagy receptor regulation|Mitophagy
      status=mapped scope=ok_for_propagation_to_go GO=[GO:0000423 mitophagy]
      rationale: The PN receptor-regulation type for mitophagy captures factors that tune receptor activity within the mitophagy pathway. This supports propagation to mitophagy while preserving that the source is a regulatory sub-role.
    • [group] Autophagy-Lysosome Pathway|Autophagy substrate selection|Autophagy receptor regulation
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a broad PN taxonomy container. The descendants mix components, regulators, context labels, and mechanistic leaves, so propagation should come only from narrower curated nodes.
    • [class] Autophagy-Lysosome Pathway|Autophagy substrate selection
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a broad substrate-selection container. GO has useful targets for specific receptor, cargo-adaptor, and selective-autophagy leaves, but this class mixes marking, recognition, receptor regulation, and unknown roles and should not propagate as one term.
    • [branch] Autophagy-Lysosome Pathway
      status=no_mapping scope= GO=[]
      rationale: Reviewed as the top-level PN branch. It is a project taxonomy umbrella rather than a direct GO assertion; all propagation must come from manually curated child nodes.

PN row 3: Ubiquitin Proteasome System | E3 ubiquitin and UBL ligases | RING | with transmembrane domain | ER, mitochondria

  • UniProt: Q96GF1
  • In branches: ER, ALP, UPS
  • Signature domains: IPR001841
  • Auxiliary domains: (none)
  • PN references (titles):
    • 19489725 / rev
  • PN-node mapping records (path + ancestors):
    • [subtype] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|RING|with transmembrane domain|ER, mitochondria
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a narrower E3-ligase architecture, component, or domain subdivision already covered by the curated parent E3 mapping. No additional direct GO mapping is needed at this node.
    • [type] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|RING|with transmembrane domain
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a narrower E3-ligase architecture, component, or domain subdivision already covered by the curated parent E3 mapping. No additional direct GO mapping is needed at this node.
    • [group] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|RING
      status=mapped scope=ok_for_propagation_to_go GO=[GO:0061630 ubiquitin protein ligase activity]
      rationale: This PN group is a catalytic ubiquitin E3 ligase bucket. The shared GO molecular-function target is ubiquitin protein ligase activity.
    • [class] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases
      status=context_only scope=too_broad_to_propagate GO=[GO:0061630 ubiquitin protein ligase activity]
      rationale: This class is a genuine E3-ligase context, but its descendants include catalytic ligases, cullin scaffolds, substrate receptors, adaptors, cofactors, regulators, and UBL modifier systems. A class-level propagation would over-annotate.
    • [branch] Ubiquitin Proteasome System
      status=no_mapping scope= GO=[]
      rationale: Reviewed as the top-level UPS branch. It is a project taxonomy umbrella rather than a direct GO assertion; UPS propagation must come from manually curated child nodes.

PN row 4: Ubiquitin Proteasome System | E3 ubiquitin and UBL ligases | idiosyncratic RING complex | membralin complex | catalytic / RING, transmembrane

  • UniProt: Q96GF1
  • In branches: ER, ALP, UPS
  • Signature domains: (none)
  • Auxiliary domains: IPR001841
  • PN references (titles):
    • 32738194
  • PN-node mapping records (path + ancestors):
    • [subtype] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|idiosyncratic RING complex|membralin complex|catalytic / RING, transmembrane
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a narrower E3-ligase architecture, component, or domain subdivision already covered by the curated parent E3 mapping. No additional direct GO mapping is needed at this node.
    • [type] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|idiosyncratic RING complex|membralin complex
      status=no_mapping scope= GO=[]
      rationale: Reviewed as a narrower E3-ligase architecture, component, or domain subdivision already covered by the curated parent E3 mapping. No additional direct GO mapping is needed at this node.
    • [group] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|idiosyncratic RING complex
      status=mapped scope=ok_for_propagation_to_go GO=[GO:0000151 ubiquitin ligase complex]
      rationale: This PN group is an E3 ligase complex bucket. The safest shared GO target is ubiquitin ligase complex membership rather than assigning catalytic activity to every subunit.
    • [class] Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases
      status=context_only scope=too_broad_to_propagate GO=[GO:0061630 ubiquitin protein ligase activity]
      rationale: This class is a genuine E3-ligase context, but its descendants include catalytic ligases, cullin scaffolds, substrate receptors, adaptors, cofactors, regulators, and UBL modifier systems. A class-level propagation would over-annotate.
    • [branch] Ubiquitin Proteasome System
      status=no_mapping scope= GO=[]
      rationale: Reviewed as the top-level UPS branch. It is a project taxonomy umbrella rather than a direct GO assertion; UPS propagation must come from manually curated child nodes.

Projected GO annotations (6)

  • GO:0036503 ERAD pathway | scope=exact | goa_status=already_in_goa_exact | from=ER proteostasis|Organelle-specific protein degradation|ER associated degradation
  • GO:0036503 ERAD pathway | scope=ok_for_propagation_to_go | goa_status=already_in_goa_exact | from=ER proteostasis|Organelle-specific protein degradation|ER associated degradation|Cytosolic handling of ERAD substrates
  • GO:0061630 ubiquitin protein ligase activity | scope=ok_for_propagation_to_go | goa_status=already_in_goa_exact | from=ER proteostasis|Organelle-specific protein degradation|ER associated degradation|Cytosolic handling of ERAD substrates|ERAD-associated RING E3 ligase
  • GO:0000423 mitophagy | scope=ok_for_propagation_to_go | goa_status=new_to_goa | from=Autophagy-Lysosome Pathway|Autophagy substrate selection|Autophagy receptor regulation|Mitophagy
  • GO:0061630 ubiquitin protein ligase activity | scope=ok_for_propagation_to_go | goa_status=already_in_goa_exact | from=Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|RING
  • GO:0000151 ubiquitin ligase complex | scope=ok_for_propagation_to_go | goa_status=new_to_goa | from=Ubiquitin Proteasome System|E3 ubiquitin and UBL ligases|idiosyncratic RING complex

Note

This file is generated from the current PROTEOSTASIS phase-1 dossier and local gene-review artifacts. Edit the source review, PN mapping, or dossier rather than this generated note when correcting the underlying curation.

📄 View Raw YAML

 
id: Q96GF1
gene_symbol: RNF185
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: >-
  RNF185 is a small (192 aa) membrane-anchored RING-type E3 ubiquitin-protein
  ligase (EC 2.3.2.27) and a close paralog of RNF5/RMA1 in the RNF5/RNF185-like
  family. It has a cytoplasmic N-terminal C3HC4 RING domain (catalytic Cys-39;
  the RING domain is responsible for ligase activity) and two C-terminal
  transmembrane helices that anchor it in membranes. RNF185 is an
  ER-membrane-resident ERAD ubiquitin ligase: it mediates the cotranslational
  ubiquitination and proteasomal degradation of the misfolded membrane protein
  CFTR (including CFTR-deltaF508), and it functions partly redundantly with RNF5
  as an E3 ligase module that is central to CFTR degradation. Its expression is
  induced by the unfolded protein response and ER stress, and it protects cells
  from ER stress-induced apoptosis; it preferentially partners with the
  ER-associated E2 enzymes UBE2J1 and UBE2J2. Beyond canonical ERAD, RNF185 has
  documented ligase-dependent regulatory roles that use distinct ubiquitin chain
  topologies. At the mitochondrial outer membrane it builds K63-linked chains on
  the Bcl-2 family protein BNIP1 to promote selective mitochondrial autophagy via
  the autophagy receptor p62, and it builds non-degradative K27-linked chains on
  the DNA sensor cGAS (CGAS) at Lys-173/Lys-384 to enhance its enzymatic activity
  and the cGAS-STING innate antiviral response. RNF185 is ubiquitously expressed;
  its core function is as an ER-membrane ERAD ubiquitin ligase.
alternative_products:
- name: '1'
  id: Q96GF1-1
- name: '2'
  id: Q96GF1-2
  sequence_note: VSP_020004
existing_annotations:
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: Phylogenetic assignment of RING-type ubiquitin ligase activity, the core molecular function of RNF185, conserved across the RNF5/RNF185 family.
    action: ACCEPT
    reason: Core molecular function; supported by multiple IDA/EXP studies (EC 2.3.2.27; RING C39A abolishes activity).
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: The RING-type zinc finger domain is responsible for E3 ubiquitin ligase activity
- term:
    id: GO:0036503
    label: ERAD pathway
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: involved_in
  review:
    summary: Phylogenetic assignment of involvement in ER-associated degradation, the core biological process of RNF185.
    action: ACCEPT
    reason: Core biological process; directly supported (CFTR ERAD) and conserved in the family.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway
- term:
    id: GO:0044390
    label: ubiquitin-like protein conjugating enzyme binding
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: RNF185 binds ubiquitin-conjugating (E2) enzymes, preferentially the ERAD E2s UBE2J1 and UBE2J2. Informative but ancillary to the ligase activity.
    action: KEEP_AS_NON_CORE
    reason: Accurately reflects E2 binding required for catalysis, but is a mechanistic subsidiary of the core ubiquitin ligase activity.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: Preferentially associates with the E2 enzymes UBE2J1 and UBE2J2
- term:
    id: GO:0004842
    label: ubiquitin-protein transferase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  qualifier: enables
  review:
    summary: ARBA electronic assignment of ubiquitin-protein transferase activity, a parent of the specific RING-type ubiquitin ligase activity.
    action: ACCEPT
    reason: Correct general molecular function; the specific GO:0061630 captures the core RING E3 ligase activity.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: EC=2.3.2.27
- term:
    id: GO:0005737
    label: cytoplasm
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  qualifier: located_in
  review:
    summary: ARBA electronic assignment of generic cytoplasmic localization; RNF185 has a cytoplasmic N-terminus but is a membrane protein.
    action: KEEP_AS_NON_CORE
    reason: Generic and imprecise; the core localization is the ER membrane. The cytoplasmic topological domain does not make the protein a soluble cytoplasmic protein.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: Cytoplasmic
- term:
    id: GO:0005741
    label: mitochondrial outer membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: Electronic transfer of mitochondrial outer membrane localization, reflecting the BNIP1/mitochondrial-autophagy study. A real but secondary localization.
    action: KEEP_AS_NON_CORE
    reason: Supported by PMID:21931693 but represents a secondary, autophagy-related localization distinct from the core ER-membrane ERAD function.
    supported_by:
    - reference_id: PMID:21931693
      supporting_text: The two C-terminal transmembrane domains of human RNF185 mediate its localization to mitochondrial outer membrane
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: IEA
  original_reference_id: GO_REF:0000002
  qualifier: located_in
  review:
    summary: InterPro-based electronic assignment of ER localization, consistent with RNF185's ER-membrane ERAD function.
    action: ACCEPT
    reason: Correct compartment; redundant with the more specific ER membrane annotations and IDA evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: Electronic transfer of ER membrane localization from the UniProt subcellular location; the core compartment for RNF185's ERAD function.
    action: ACCEPT
    reason: Correct core localization; supported experimentally (EXP, PMID:27485036).
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Endoplasmic reticulum membrane {ECO:0000269|PubMed:24019521, ECO:0000269|PubMed:27485036}'
- term:
    id: GO:0006511
    label: ubiquitin-dependent protein catabolic process
  evidence_type: IEA
  original_reference_id: GO_REF:0000002
  qualifier: involved_in
  review:
    summary: InterPro-based electronic assignment of ubiquitin-dependent protein catabolism, a parent process consistent with RNF185's ERAD role.
    action: ACCEPT
    reason: Correct but generic; the specific GO:0036503 (ERAD pathway) better captures the core biological role.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: unfolded proteins that accumulate in the ER are transported to the cytosol for ubiquitin-proteasome-mediated degradation
- term:
    id: GO:0036503
    label: ERAD pathway
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  qualifier: involved_in
  review:
    summary: ARBA machine-learning electronic assignment of the ERAD pathway, redundant with the experimentally supported core process.
    action: ACCEPT
    reason: Correct core biological process; redundant with IMP/IGI and IBA evidence.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway
- term:
    id: GO:0044322
    label: endoplasmic reticulum quality control compartment
  evidence_type: IEA
  original_reference_id: GO_REF:0000108
  qualifier: located_in
  review:
    summary: Inter-ontology logical inference placing RNF185 in the ER quality control compartment, a plausible localization derived from its ERAD role.
    action: KEEP_AS_NON_CORE
    reason: Plausible localization inferred from the ERAD/ERQC link, but not directly demonstrated; the core localization annotation is ER membrane.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0046872
    label: metal ion binding
  evidence_type: IEA
  original_reference_id: GO_REF:0000002
  qualifier: enables
  review:
    summary: InterPro-based assignment of metal (zinc) ion binding by the RING domain, a structural requirement for the ligase fold.
    action: KEEP_AS_NON_CORE
    reason: Accurate structural feature of the RING domain but subsidiary to the informative ubiquitin ligase activity; not a standalone core function.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: RING-type
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: IEA
  original_reference_id: GO_REF:0000120
  qualifier: enables
  review:
    summary: Combined automated electronic assignment of the core RING E3 ligase activity, consistent with EC 2.3.2.27 and experimental evidence.
    action: ACCEPT
    reason: Correct core molecular function; redundant with IDA/EXP evidence.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: EC=2.3.2.27
- term:
    id: GO:1904380
    label: endoplasmic reticulum mannose trimming
  evidence_type: IEA
  original_reference_id: GO_REF:0000117
  qualifier: involved_in
  review:
    summary: ARBA machine-learning assignment of ER mannose trimming. RNF185 is a ubiquitin ligase and does not trim mannose; this is pathway-adjacency over-annotation.
    action: REMOVE
    reason: RNF185 has no glycosidase/mannosidase activity; mannose trimming is performed by EDEM/ER mannosidases. This electronic inference is biologically incorrect.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: E3 ubiquitin-protein ligase that regulates selective
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19549727
  qualifier: enables
  review:
    summary: E2 ubiquitin-conjugating enzyme interaction network capturing RNF185 binding to UBE2D/UBE2E enzymes. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Real E2 interactions relevant to catalysis but bare protein binding is uninformative; E2 binding is captured by GO:0044390.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Q96GF1; P51668: UBE2D1'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25416956
  qualifier: enables
  review:
    summary: Proteome-scale interactome map capturing RNF185 partners including RNF5. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Q96GF1; Q99942: RNF5'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:28514442
  qualifier: enables
  review:
    summary: Interactome-community study capturing RNF185 binding to TMBIM6/BI-1. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Q96GF1; P55061: TMBIM6'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:31515488
  qualifier: enables
  review:
    summary: Interaction captured in a study of variant-driven interactome disruption. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interaction; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Q96GF1; Q969T4: UBE2E3'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:32296183
  qualifier: enables
  review:
    summary: Binary interactome reference map capturing many RNF185 membrane-protein partners. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Q96GF1; P55061: TMBIM6'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:33961781
  qualifier: enables
  review:
    summary: Cell-specific interactome capturing RNF185 partners including RNF5. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'Q96GF1; Q99942: RNF5'
- term:
    id: GO:0055085
    label: transmembrane transport
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-382556
  qualifier: involved_in
  review:
    summary: Reactome pathway-level annotation (ABC-family transport). RNF185 is a ubiquitin ligase, not a transporter; this reflects CFTR/ABC pathway-context bleed-through.
    action: REMOVE
    reason: RNF185 does not mediate transmembrane transport; it is an E3 ligase acting on transporter substrates such as CFTR. The transport annotation is a pathway-adjacency artifact.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway
- term:
    id: GO:0016567
    label: protein ubiquitination
  evidence_type: IEA
  original_reference_id: GO_REF:0000041
  qualifier: involved_in
  review:
    summary: UniPathway-derived general protein ubiquitination process, a parent of the specific ERAD ubiquitination RNF185 performs.
    action: KEEP_AS_NON_CORE
    reason: Correct but generic; the specific ERAD pathway and K63/K27 ubiquitination annotations better capture the role.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: 'PATHWAY: Protein modification; protein ubiquitination.'
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866546
  qualifier: enables
  review:
    summary: Reactome curation of RNF185 (with RNF5) ubiquitinating misfolded CFTR; captures the core ligase activity.
    action: ACCEPT
    reason: Correct core molecular function in the CFTR ERAD reaction.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8867288
  qualifier: enables
  review:
    summary: Reactome curation of the ERAD E3 ligase ubiquitinating an unfolded glycoprotein substrate; captures the core ligase activity.
    action: ACCEPT
    reason: Correct core molecular function.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: The RING-type zinc finger domain is responsible for E3 ubiquitin ligase activity
- term:
    id: GO:0005741
    label: mitochondrial outer membrane
  evidence_type: EXP
  original_reference_id: PMID:21931693
  qualifier: located_in
  review:
    summary: Experimental localization to the mitochondrial outer membrane, where RNF185 ubiquitinates BNIP1 during mitochondrial autophagy. A real but secondary localization.
    action: KEEP_AS_NON_CORE
    reason: Directly supported (PMID:21931693) but a secondary compartment for the autophagy role, not the core ER ERAD function.
    supported_by:
    - reference_id: PMID:21931693
      supporting_text: The two C-terminal transmembrane domains of human RNF185 mediate its localization to mitochondrial outer membrane
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: EXP
  original_reference_id: PMID:27485036
  qualifier: located_in
  review:
    summary: Experimental evidence that RNF185 localizes to the ER membrane; the core compartment for its ERAD ligase activity.
    action: ACCEPT
    reason: Core localization with direct experimental support.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: EXP
  original_reference_id: PMID:21931693
  qualifier: enables
  review:
    summary: Experimental demonstration of RNF185 ubiquitin ligase activity (K63-linked polyubiquitination of BNIP1). Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental support.
    supported_by:
    - reference_id: PMID:21931693
      supporting_text: human RNF185 is a mitochondrial ubiquitin E3 ligase that regulates selective mitochondrial autophagy in cultured cells
- term:
    id: GO:0006511
    label: ubiquitin-dependent protein catabolic process
  evidence_type: IMP
  original_reference_id: PMID:24019521
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence that RNF185 drives ubiquitin-dependent degradation of CFTR. Consistent with the core ERAD/proteasomal degradation role.
    action: ACCEPT
    reason: Supported by RNF185 depletion stabilizing CFTR; consistent with the core ERAD function.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: its silencing stabilizes CFTR proteins
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: IDA
  original_reference_id: PMID:24019521
  qualifier: enables
  review:
    summary: Direct evidence of RNF185 RING-dependent ubiquitin ligase activity controlling CFTR stability. Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental (IDA) support.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner
- term:
    id: GO:0045089
    label: positive regulation of innate immune response
  evidence_type: IDA
  original_reference_id: PMID:28273161
  qualifier: involved_in
  review:
    summary: Direct evidence that RNF185 positively regulates the cGAS-mediated innate immune response. A real but secondary, immunity-specific role.
    action: KEEP_AS_NON_CORE
    reason: Well supported (PMID:28273161) but a secondary moonlighting role distinct from the core ER ERAD function.
    supported_by:
    - reference_id: PMID:28273161
      supporting_text: RNF185 specifically catalyzed the K27-linked poly-ubiquitination of cGAS, which promoted its enzymatic activity
- term:
    id: GO:1904380
    label: endoplasmic reticulum mannose trimming
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-901032
  qualifier: involved_in
  review:
    summary: Reactome ERQC pathway annotation. RNF185 does not perform mannose trimming; this is pathway-adjacency over-annotation.
    action: REMOVE
    reason: RNF185 is an E3 ligase with no mannosidase activity; ER mannose trimming is carried out by ER mannosidases/EDEMs. The annotation conflates pathway membership with direct activity.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: E3 ubiquitin-protein ligase that regulates selective
- term:
    id: GO:0044314
    label: protein K27-linked ubiquitination
  evidence_type: IDA
  original_reference_id: PMID:28273161
  qualifier: involved_in
  review:
    summary: Direct evidence that RNF185 builds non-degradative K27-linked polyubiquitin chains on cGAS, enhancing its activity. A secondary, immunity-related activity.
    action: KEEP_AS_NON_CORE
    reason: Directly demonstrated K27-linked ubiquitination, but in the context of the secondary cGAS-STING immune role rather than the core ERAD function. K27-linked chain building by RNF185 is corroborated on additional substrates (EBOV GP1,2, PMID:36224200; TUFM, PMID:38084826).
    supported_by:
    - reference_id: PMID:28273161
      supporting_text: RNF185 specifically catalyzed the K27-linked poly-ubiquitination of cGAS
    - reference_id: PMID:36224200
    - reference_id: PMID:38084826
- term:
    id: GO:0051607
    label: defense response to virus
  evidence_type: IDA
  original_reference_id: PMID:28273161
  qualifier: involved_in
  review:
    summary: Direct evidence that RNF185 contributes to antiviral defense via the cGAS-STING pathway. A real but secondary, immunity-specific role.
    action: KEEP_AS_NON_CORE
    reason: Well supported but a secondary moonlighting role distinct from the core ER ERAD function.
    supported_by:
    - reference_id: PMID:28273161
      supporting_text: the ER ubiquitin ligase RNF185 interacted with cGAS during HSV-1 infection
- term:
    id: GO:0060340
    label: positive regulation of type I interferon-mediated signaling pathway
  evidence_type: IDA
  original_reference_id: PMID:28273161
  qualifier: involved_in
  review:
    summary: Direct evidence that RNF185 positively regulates type I interferon signaling via cGAS. A real but secondary, immunity-specific role.
    action: KEEP_AS_NON_CORE
    reason: Well supported but a secondary moonlighting role distinct from the core ER ERAD function.
    supported_by:
    - reference_id: PMID:28273161
      supporting_text: Ectopic-expression or knockdown of RNF185 respectively enhanced or impaired the IRF3-responsive gene expression
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: IDA
  original_reference_id: PMID:28273161
  qualifier: enables
  review:
    summary: Direct evidence of RNF185 ubiquitin ligase activity (K27-linked ubiquitination of cGAS; RING-dependent). Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental support; C39A/C79A abolishes activity.
    supported_by:
    - reference_id: PMID:28273161
      supporting_text: RNF185 specifically catalyzed the K27-linked poly-ubiquitination of cGAS
- term:
    id: GO:0044877
    label: protein-containing complex binding
  evidence_type: IPI
  original_reference_id: PMID:24019521
  qualifier: enables
  review:
    summary: RNF185 binds a protein complex (ERAD machinery) in the CFTR degradation study. More informative than bare protein binding but still ancillary.
    action: KEEP_AS_NON_CORE
    reason: Reflects association with the ERAD E3 module/machinery; supportive of, but subsidiary to, the core ligase and ERAD-process annotations.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: IDA
  original_reference_id: PMID:27485036
  qualifier: located_in
  review:
    summary: Direct evidence for ER localization, consistent with RNF185's ER-membrane ERAD function.
    action: ACCEPT
    reason: IDA-supported ER localization agrees with the documented ER-membrane site of action.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0043130
    label: ubiquitin binding
  evidence_type: IDA
  original_reference_id: PMID:27485036
  qualifier: enables
  review:
    summary: Direct evidence that RNF185 binds ubiquitin, consistent with its in vitro autoubiquitination/ligase activity.
    action: ACCEPT
    reason: Supported by the in vitro autoubiquitination assays; reflects ubiquitin handling integral to ligase function.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: in vitro autoubiquitination activity
- term:
    id: GO:0051865
    label: protein autoubiquitination
  evidence_type: IDA
  original_reference_id: PMID:27485036
  qualifier: involved_in
  review:
    summary: RNF185 is autoubiquitinated in vitro, a common property of RING E3 ligases reflecting their catalytic activity.
    action: KEEP_AS_NON_CORE
    reason: Supported by in vitro autoubiquitination assays but a secondary property of the ligase, not its core substrate-directed function.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: in vitro autoubiquitination activity
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: IDA
  original_reference_id: PMID:27485036
  qualifier: enables
  review:
    summary: Direct evidence of RNF185 E3 activity, with E3-activity-dependent resistance to ER stress. Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental support; RING C39S decreases activity.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: RNF185, but not CGRRF1 and RNF19B, exhibited significant resistance to ER stressor in an E3 activity-dependent manner
- term:
    id: GO:1904294
    label: positive regulation of ERAD pathway
  evidence_type: IMP
  original_reference_id: PMID:27485036
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence that RNF185 positively regulates ERAD, protecting cells from ER stress in an E3-activity-dependent manner. Core biological process.
    action: ACCEPT
    reason: Core biological process; RNF185 is an ER-stress-induced ERAD E3 that confers ER stress resistance.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: RNF185, but not CGRRF1 and RNF19B, exhibited significant resistance to ER stressor in an E3 activity-dependent manner
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:21931693
  qualifier: enables
  review:
    summary: Interaction with BNIP1, a functionally important RNF185 substrate at the mitochondrial outer membrane. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Records the real RNF185-BNIP1 interaction but bare protein binding is uninformative.
    supported_by:
    - reference_id: PMID:21931693
      supporting_text: We further identified the Bcl-2 family protein BNIP1 as one of the substrates for RNF185
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866542
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization within CFTR ERAD reactions. Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866546
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (RNF5/RNF185 ubiquitinate misfolded CFTR). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866551
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (CFTR binds ERAD machinery). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866854
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (CFTR F508del translocation). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866856
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (RNF5/RNF185 ubiquitinate CFTR F508del). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866857
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (CFTR F508del binds ERAD machinery). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8867288
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization in an ERAD ubiquitination reaction. Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-9931264
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (CD274/PD-L1 ERAD transport). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-9931298
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (ubiquitination of CD274 by ERAD complex). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-9931313
  qualifier: located_in
  review:
    summary: Reactome curation of RNF185 ER membrane localization (p-CD274 binds ERAD complex). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:27485036
      supporting_text: All were partially localised to the ER
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:24019521
  qualifier: enables
  review:
    summary: Interaction captured in the RNF185/RNF5 CFTR ERAD study. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Real interaction within the ERAD module but bare protein binding is uninformative.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: IDA
  original_reference_id: PMID:24019521
  qualifier: located_in
  review:
    summary: Direct evidence for ER localization in the CFTR ERAD study. Core compartment.
    action: ACCEPT
    reason: IDA-supported ER localization agrees with the documented ER-membrane ERAD function.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: RNF185 is a RING domain-containing polypeptide homologous to RNF5
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: NAS
  original_reference_id: PMID:24019521
  qualifier: located_in
  review:
    summary: Author-stated ER membrane localization in the CFTR ERAD study. Core compartment.
    action: ACCEPT
    reason: Consistent with experimentally supported ER-membrane localization.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: RNF185 is a RING domain-containing polypeptide homologous to RNF5
- term:
    id: GO:0036503
    label: ERAD pathway
  evidence_type: IGI
  original_reference_id: PMID:24019521
  qualifier: involved_in
  review:
    summary: Genetic-interaction evidence (with RNF5) that RNF185 drives ERAD of CFTR; co-depletion blocks CFTR-deltaF508 degradation. Core biological process.
    action: ACCEPT
    reason: Core biological process with direct experimental support from the RNF5/RNF185 co-depletion experiments.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: simultaneous depletion of RNF5 and RNF185 profoundly blocks CFTRΔF508 degradation not only during translation but also after synthesis is complete
- term:
    id: GO:0044390
    label: ubiquitin-like protein conjugating enzyme binding
  evidence_type: IPI
  original_reference_id: PMID:24019521
  qualifier: enables
  review:
    summary: RNF185 binds the ERAD E2 enzymes UBE2J1/UBE2J2, demonstrated in the CFTR ERAD study. Informative but ancillary to ligase activity.
    action: KEEP_AS_NON_CORE
    reason: Reflects E2 (UBE2J1/UBE2J2) binding required for catalysis, but is a mechanistic subsidiary of the core ligase activity.
    supported_by:
    - reference_id: file:human/RNF185/RNF185-uniprot.txt
      supporting_text: Preferentially associates with the E2 enzymes UBE2J1 and UBE2J2
- term:
    id: GO:0051865
    label: protein autoubiquitination
  evidence_type: IDA
  original_reference_id: PMID:24019521
  qualifier: involved_in
  review:
    summary: RNF185 autoubiquitinates, a common RING E3 property reflecting its catalytic activity.
    action: KEEP_AS_NON_CORE
    reason: Supported but a secondary property of the ligase, not its core substrate-directed function.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner
references:
- id: GO_REF:0000002
  title: Gene Ontology annotation through association of InterPro records with GO terms
  findings: []
- id: GO_REF:0000033
  title: Annotation inferences using phylogenetic trees
  findings: []
- id: GO_REF:0000041
  title: Gene Ontology annotation based on UniPathway vocabulary mapping
  findings: []
- id: GO_REF:0000044
  title: Gene Ontology annotation based on UniProtKB/Swiss-Prot Subcellular Location vocabulary mapping
  findings: []
- id: GO_REF:0000108
  title: Automatic assignment of GO terms using logical inference, based on inter-ontology links
  findings: []
- id: GO_REF:0000117
  title: Electronic Gene Ontology annotations created by ARBA machine learning models
  findings: []
- id: GO_REF:0000120
  title: Combined Automated Annotation using Multiple IEA Methods
  findings: []
- id: PMID:19549727
  title: Analysis of the human E2 ubiquitin conjugating enzyme protein interaction network.
  findings: []
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: E2 interaction network; documents RNF185 binding to multiple UBE2D/UBE2E enzymes relevant to its catalytic cycle.
- id: PMID:21931693
  title: RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates autophagy through interaction with BNIP1.
  findings:
  - statement: RNF185 is a mitochondrial outer membrane E3 ligase that builds K63-linked polyubiquitin chains on BNIP1 to promote selective mitochondrial autophagy via recruitment of the autophagy receptor p62.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Original RNF185 characterization; establishes ligase activity and the non-core mitochondrial-autophagy/BNIP1 role and mitochondrial OM localization.
- id: PMID:24019521
  title: RNF185 is a novel E3 ligase of endoplasmic reticulum-associated degradation (ERAD) that targets cystic fibrosis transmembrane conductance regulator (CFTR).
  findings:
  - statement: RNF185 is an ER-membrane RING E3 that controls cotranslational ubiquitination and proteasomal degradation of CFTR/CFTR-deltaF508 in a RING- and proteasome-dependent manner; RNF185 and RNF5 form a partly redundant E3 module central to CFTR degradation, preferentially using E2 enzymes UBE2J1/UBE2J2.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Full text available; establishes the core ER-membrane ERAD/CFTR role and RNF5/RNF185 redundancy; source of IMP/IGI/IDA annotations.
- id: PMID:25416956
  title: A proteome-scale map of the human interactome network.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput interactome; source of multiple bare protein binding annotations.
- id: PMID:27485036
  title: Genome-wide identification and gene expression profiling of ubiquitin ligases for endoplasmic reticulum protein degradation.
  findings:
  - statement: RNF185 is an ER-stress/UPR-induced ER-localized RING E3 with in vitro autoubiquitination activity that confers ER stress resistance in an E3-activity-dependent manner; the RING domain (Cys-39) is responsible for ligase activity.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Full text available; establishes ER localization, ER-stress induction, E3-activity-dependent ER stress resistance (positive regulation of ERAD).
- id: PMID:28273161
  title: The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated innate immune response.
  findings:
  - statement: ER-resident RNF185 interacts with cGAS during HSV-1 infection and catalyzes non-degradative K27-linked polyubiquitination of cGAS, enhancing its enzymatic activity and the cGAS-STING type I interferon antiviral response; RING residues Cys-39/Cys-79 are required.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Full text available; establishes the non-core cGAS-STING immunity role and K27-linked ubiquitination; source of immune-process annotations.
- id: PMID:28514442
  title: Architecture of the human interactome defines protein communities and disease networks.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput interactome-community study; source of a bare protein binding annotation (TMBIM6).
- id: PMID:31515488
  title: Extensive disruption of protein interactions by genetic variants across the allele frequency spectrum in human populations.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput variant-interactome study; source of a bare protein binding annotation.
- id: PMID:32296183
  title: A reference map of the human binary protein interactome.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Binary interactome reference map; source of multiple bare protein binding annotations.
- id: PMID:33961781
  title: Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Cell-specific interactome; source of bare protein binding annotations (including RNF5).
- id: Reactome:R-HSA-382556
  title: ABC-family proteins mediated transport
  findings: []
- id: Reactome:R-HSA-8866542
  title: VCP-catalyzed ATP hydrolysis promotes the translocation of misfolded CFTR into the cytosol
  findings: []
- id: Reactome:R-HSA-8866546
  title: RNF5 and RNF185 ubiquitinate misfolded CFTR
  findings: []
- id: Reactome:R-HSA-8866551
  title: CFTR binds components of the ERAD machinery for ubiquitination and degradation
  findings: []
- id: Reactome:R-HSA-8866854
  title: VCP-catalyzed ATP hydrolysis promotes the translocation of CFTR F508del into the cytosol
  findings: []
- id: Reactome:R-HSA-8866856
  title: RNF5 and RNF185 ubiquitinate CFTR F508del
  findings: []
- id: Reactome:R-HSA-8866857
  title: CFTR F508del binds components of the ERAD machinery for ubiquitination and degradation
  findings: []
- id: Reactome:R-HSA-8867288
  title: OS9:SEL1:ERAD E3 ligase:DERL2 ubiquitinates unfolded protein:(GlcNAc)2 (Man)9-5
  findings: []
- id: Reactome:R-HSA-901032
  title: ER Quality Control Compartment (ERQC)
  findings: []
- id: Reactome:R-HSA-9931264
  title: Active transport of ubiquitinated CD274 from ER to cytosol
  findings: []
- id: Reactome:R-HSA-9931298
  title: Ubiquitination of CD274 by ERAD complex
  findings: []
- id: Reactome:R-HSA-9931313
  title: p-S195-CD274 binds ERAD complex
  findings: []
- id: PMID:36224200
  title: RNF185 regulates proteostasis in Ebolavirus infection by crosstalk between the calnexin cycle, ERAD, and reticulophagy.
  findings:
  - statement: The ER ubiquitin ligase RNF185 polyubiquitinates misfolded Ebolavirus glycoprotein GP on Lys-673 via K27-linked chains, recruiting SQSTM1/p62 to divert GP to ATG3/ATG5-dependent reticulophagy/ERLAD (lysosomal) rather than proteasomal degradation.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: PubMed-verified (PMID:36224200 = Nat Commun 2022, DOI 10.1038/s41467-022-33805-9). Extends the RNF185 non-degradative K27-linkage repertoire to a viral ERAD/reticulophagy substrate; full text not in cache, so no supporting_text added.
- id: PMID:38084826
  title: Senecavirus A induces mitophagy to promote self-replication through direct interaction of 2C protein with K27-linked ubiquitinated TUFM catalyzed by RNF185.
  findings:
  - statement: RNF185 catalyzes K27-linked polyubiquitination of the mitochondrial translation elongation factor TUFM (via RNF185 transmembrane domain 1), enabling SQSTM1/p62 recognition and mitophagy that is exploited by Senecavirus A to promote replication.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: PubMed-verified (PMID:38084826 = Autophagy 2024, DOI 10.1080/15548627.2023.2293442). Adds a new K27-linked RNF185 substrate (TUFM) and links RNF185 to mitophagy in a viral context; full text not in cache, so no supporting_text added.
- id: PMID:39353943
  title: Tapasin assembly surveillance by the RNF185/Membralin ubiquitin ligase complex regulates MHC-I surface expression.
  findings:
  - statement: RNF185 forms an ERAD complex with Membralin (MBRL/TMEM259) that recognizes unassembled Tapasin (a peptide-loading-complex component) and targets it for ubiquitin-dependent degradation; loss of RNF185/Membralin raises Tapasin steady-state levels and increases surface MHC-I, defining an ERAD-based control of antigen presentation.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: PubMed-verified (PMID:39353943 = Nat Commun 2024, DOI 10.1038/s41467-024-52772-x). Major new finding identifying the RNF185/Membralin ERAD complex and a new physiological substrate (Tapasin) that links RNF185 to MHC-I antigen presentation; full text not in cache, so no supporting_text added to annotations.
core_functions:
- description: RING-type E3 ubiquitin ligase anchored in the ER membrane that mediates cotranslational ubiquitination of misfolded membrane-protein clients (notably CFTR and CFTR-deltaF508) for proteasomal degradation via the ERAD pathway, acting partly redundantly with RNF5 and preferentially using the E2 enzymes UBE2J1/UBE2J2.
  molecular_function:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  locations:
  - id: GO:0005789
    label: endoplasmic reticulum membrane
  supported_by:
  - reference_id: PMID:24019521
    supporting_text: RNF185 controls the stability of CFTR and of the CFTRΔF508 mutant in a RING- and proteasome-dependent manner
  - reference_id: PMID:24019521
    supporting_text: identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation
  directly_involved_in:
  - id: GO:0036503
    label: ERAD pathway
- description: ER-stress/UPR-induced ER-membrane ubiquitin ligase that positively regulates ERAD and confers resistance to ER stress-induced apoptosis in an E3-activity-dependent manner.
  molecular_function:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  locations:
  - id: GO:0005789
    label: endoplasmic reticulum membrane
  supported_by:
  - reference_id: PMID:27485036
    supporting_text: RNF185, but not CGRRF1 and RNF19B, exhibited significant resistance to ER stressor in an E3 activity-dependent manner
  directly_involved_in:
  - id: GO:1904294
    label: positive regulation of ERAD pathway
proposed_new_terms: []
suggested_questions:
- question: How is RNF185 partitioned between the ER membrane (ERAD/CFTR) and the mitochondrial outer membrane (BNIP1/mitophagy), and what governs this dual localization and substrate choice?
- question: To what extent are the RNF5 and RNF185 ERAD substrate repertoires overlapping versus distinct, and is their redundancy substrate-specific?
suggested_experiments:
- description: Reconstitute CFTR-deltaF508 ubiquitination in vitro with purified RNF185, RNF5, UBE2J1/UBE2J2 and Derlin cofactors to compare paralog activity and map chain linkages, and perform RNF5/RNF185 single and double knockouts with quantitative ubiquitinome profiling to define overlapping vs distinct ERAD substrates.
- description: Use proximity labeling (BioID/TurboID) of ER-anchored vs mitochondria-anchored RNF185 to map compartment-specific interactomes and test which substrates (CFTR, BNIP1, cGAS) depend on each localization.