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 RNF5, conserved across the RNF5/RNF185 family. Reason: Core molecular function; supported experimentally (EC 2.3.2.27; C42S abolishes activity) and across the family. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
GO:0036503 ERAD pathway	IBA GO_REF:0000033	ACCEPT	Summary: Phylogenetic assignment of involvement in ER-associated degradation, the core biological process of RNF5 as an ER-anchored ERAD ligase. Reason: Core biological process; directly supported by IMP evidence (CFTR/CFTR-deltaF508 degradation) and conserved in the family. Supporting Evidence: PMID:24019521 Cystic fibrosis transmembrane conductance regulator (CFTR) is one ERAD substrate targeted to co-translational degradation by the E3 ligase RNF5/RMA1
GO:0044390 ubiquitin-like protein conjugating enzyme binding	IBA GO_REF:0000033	KEEP AS NON CORE	Summary: RNF5 binds ubiquitin-conjugating (E2) enzymes, including the UBE2D/UbcH5 family and UBE2N/Ubc13, as part of its catalytic cycle. This is informative but ancillary to the ligase activity itself. Reason: Accurately reflects E2 binding (UBE2D1/UBE2D2, UBE2N) required for catalysis, but is a mechanistic subsidiary of the core ubiquitin ligase activity. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt May function together with E2 ubiquitin-conjugating enzymes UBE2D1/UBCH5A and UBE2D2/UBC4
GO:0005783 endoplasmic reticulum	IEA GO_REF:0000002	ACCEPT	Summary: InterPro-based electronic assignment of ER localization, consistent with RNF5's ER-membrane site of ERAD function. Reason: Correct compartment; redundant with the more specific ER membrane annotations and IDA (HPA) evidence. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Endoplasmic reticulum membrane {ECO:0000269\|PubMed:19285439}
GO:0005789 endoplasmic reticulum membrane	IEA GO_REF:0000044	ACCEPT	Summary: Electronic transfer of ER membrane localization from the UniProt subcellular location; this is the core compartment for RNF5's ERAD ligase function. Reason: Correct core localization; RNF5 is a tail-anchored ER membrane protein, supported experimentally (EXP, PMID:19285439). Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Endoplasmic reticulum membrane {ECO:0000269\|PubMed:19285439}
GO:0005886 plasma membrane	IEA GO_REF:0000044	KEEP AS NON CORE	Summary: Electronic transfer of plasma membrane localization from the original cloning study (PMID:9533025). A real but secondary localization not tied to the core ERAD function. Reason: Reported in the early cloning paper but the functionally dominant compartment for RNF5 is the ER membrane; plasma membrane pool is peripheral. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Cell membrane {ECO:0000269\|PubMed:9533025}
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 RNF5's ERAD role. Reason: Correct but generic; the specific GO:0036503 (ERAD pathway) better captures the core biological role. Supporting Evidence: PMID:24019521 degraded by the ubiquitin-proteasome pathway through a process called ER-associated degradation (ERAD)
GO:0031966 mitochondrial membrane	IEA GO_REF:0000044	KEEP AS NON CORE	Summary: Electronic transfer of mitochondrial membrane localization, reflecting the antiviral STING1/MITA study in which RNF5 acts at mitochondria. A real but secondary, immunity-related localization. Reason: Supported by PMID:19285439 (MITA ubiquitination at mitochondria) but represents a secondary, context-specific role distinct from the core ER-membrane ERAD function. Supporting Evidence: PMID:19285439 virus-induced ubiquitination and degradation of MITA by RNF5 occurred at the mitochondria
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 and IBA evidence. Supporting Evidence: PMID:24019521 Cystic fibrosis transmembrane conductance regulator (CFTR) is one ERAD substrate targeted to co-translational degradation by the E3 ligase RNF5/RMA1
GO:0044322 endoplasmic reticulum quality control compartment	IEA GO_REF:0000108	KEEP AS NON CORE	Summary: Inter-ontology logical inference placing RNF5 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:24019521 hosts a machinery called ERQC for ER quality control
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 EXP/IBA evidence. Supporting Evidence: file:human/RNF5/RNF5-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. RNF5 is a ubiquitin ligase and does not trim mannose; this is a pathway-adjacency over-annotation propagated from the ERAD/ERQC context. Reason: RNF5 has no glycosidase/mannosidase activity; mannose trimming is performed by EDEM/ER mannosidases, not by the E3 ligase. This electronic inference is biologically incorrect. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
GO:0005515 protein binding	IPI PMID:12861019 RNF5, a RING finger protein that regulates cell motility by ...	KEEP AS NON CORE	Summary: IntAct interaction with paxillin (PXN), a functionally meaningful RNF5 substrate, but the bare protein binding term is uninformative. Reason: Records a real substrate interaction (PXN) but bare protein binding is uninformative per curation guidelines. Supporting Evidence: PMID:12861019 the human homologue of RNF5 associates with the amino-terminal domain of paxillin
GO:0005515 protein binding	IPI PMID:14667819 Analysis of a high-throughput yeast two-hybrid system and it...	KEEP AS NON CORE	Summary: High-throughput yeast two-hybrid interactions (e.g. ABHD16A, UBE2 enzymes). Bare protein binding is uninformative. Reason: Records real IntAct interactions but bare protein binding is uninformative and not a core function. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; O95870: ABHD16A
GO:0005515 protein binding	IPI PMID:16901789 Sequential quality-control checkpoints triage misfolded cyst...	KEEP AS NON CORE	Summary: Interaction with CFTR captured in the sequential-triage ERAD study; CFTR is a key RNF5 ERAD substrate. Bare protein binding is uninformative. Reason: Records the functionally important RNF5-CFTR interaction, but bare protein binding is uninformative; the substrate relationship is captured by the ERAD process annotations. Supporting Evidence: PMID:16901789 an ER membrane-associated ubiquitin ligase complex containing the E3 RMA1, the E2 Ubc6e, and Derlin-1
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 RNF5 binding to multiple UBE2D/UBE2E/UBE2W E2s. 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/RNF5/RNF5-uniprot.txt Q99942; P51668: UBE2D1
GO:0005515 protein binding	IPI PMID:20152160 Crystal structure of UbcH5b~ubiquitin intermediate: insight ...	KEEP AS NON CORE	Summary: Interaction with the E2 UBE2D2/UbcH5b from a structural study of the E2~Ub conjugate. Bare protein binding is uninformative. Reason: Real E2 interaction but bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; P62837: UBE2D2
GO:0005515 protein binding	IPI PMID:25416956 A proteome-scale map of the human interactome network.	KEEP AS NON CORE	Summary: Large-scale interactome map capturing numerous RNF5 partners (including SLC transporters, RNF185, SEC22A). Bare protein binding is uninformative. Reason: High-throughput interactome; bare protein binding is uninformative and not a core function. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q96GF1: RNF185
GO:0005515 protein binding	IPI PMID:25759021 Regulation of glutamine carrier proteins by RNF5 determines ...	KEEP AS NON CORE	Summary: Interactions with glutamine carrier proteins (SLC1A5, SLC38A2) relevant to RNF5's role in breast cancer ER-stress response. Bare protein binding is uninformative. Reason: Records real substrate/carrier interactions but bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q15758: SLC1A5
GO:0005515 protein binding	IPI PMID:26618866 ∆F508 CFTR interactome remodelling promotes rescue of cystic...	KEEP AS NON CORE	Summary: Interaction with CFTR from a deltaF508 CFTR interactome-remodeling study. Bare protein binding is uninformative. Reason: Real CFTR interaction relevant to ERAD but bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; P13569: CFTR
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/RNF5/RNF5-uniprot.txt Q99942; 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 RNF5 membrane-protein partners. Bare protein binding is uninformative. Reason: High-throughput interactome; bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q9Y5U4: INSIG2
GO:0005515 protein binding	IPI PMID:32814053 Interactome Mapping Provides a Network of Neurodegenerative ...	KEEP AS NON CORE	Summary: Interactions (e.g. OPTN, UBE2K) from a neurodegenerative-disease interactome study. Bare protein binding is uninformative. Reason: High-throughput interaction; bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q96CV9: OPTN
GO:0005515 protein binding	IPI PMID:33961781 Dual proteome-scale networks reveal cell-specific remodeling...	KEEP AS NON CORE	Summary: Cell-specific interactome capturing RNF5 partners including RNF185 and RHBDD1. Bare protein binding is uninformative. Reason: High-throughput interactome; bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q96GF1: RNF185
GO:0005515 protein binding	IPI PMID:36012204 Differential CFTR-Interactome Proximity Labeling Procedures ...	KEEP AS NON CORE	Summary: Proximity-labeling interaction with CFTR identifying enrichment in SLC transporters. Bare protein binding is uninformative. Reason: Real CFTR/SLC interactions but bare protein binding is uninformative. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; P13569: CFTR
GO:0042802 identical protein binding	IPI PMID:25416956 A proteome-scale map of the human interactome network.	KEEP AS NON CORE	Summary: RNF5 self-interaction captured in a proteome-scale interactome. Reflects homo-association seen in screens. Reason: Documents RNF5 self-association (IntAct EBI-348482 with itself) but is peripheral to the core ligase function. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q99942: RNF5
GO:0042802 identical protein binding	IPI PMID:32296183 A reference map of the human binary protein interactome.	KEEP AS NON CORE	Summary: RNF5 self-interaction captured in the binary interactome reference map. Reason: Documents RNF5 self-association but is peripheral to the core ligase function. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Q99942; Q99942: RNF5
GO:0016020 membrane	IEA GO_REF:0000107	KEEP AS NON CORE	Summary: Ortholog-based electronic assignment of generic membrane localization, a parent of the specific ER membrane localization. Reason: Correct but generic; subsumed by the more specific ER membrane localization. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Multi-pass membrane protein
GO:0055085 transmembrane transport	TAS Reactome:R-HSA-382556	REMOVE	Summary: Reactome pathway-level annotation (ABC-family transport). RNF5 is a ubiquitin ligase, not a transporter; this reflects pathway context (CFTR/ABC transport) bleed-through. Reason: RNF5 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, not a direct function. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
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-associated ubiquitination RNF5 performs. Reason: Correct but generic; the specific ERAD pathway and K48/K63 ubiquitination annotations better capture the role. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt PATHWAY: Protein modification; protein ubiquitination.
GO:0061630 ubiquitin protein ligase activity	TAS Reactome:R-HSA-8866546	ACCEPT	Summary: Reactome curation of RNF5 (with RNF185) 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/RNF5/RNF5-uniprot.txt Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
GO:0005783 endoplasmic reticulum	IDA GO_REF:0000052	ACCEPT	Summary: Direct immunofluorescence (HPA) evidence for ER localization, consistent with RNF5's ER-membrane ERAD function. Reason: IDA-supported ER localization agrees with the documented ER-membrane site of action. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Endoplasmic reticulum membrane {ECO:0000269\|PubMed:19285439}
GO:0005789 endoplasmic reticulum membrane	EXP PMID:19285439 The ubiquitin ligase RNF5 regulates antiviral responses by m...	ACCEPT	Summary: Experimental evidence that RNF5 localizes to the ER membrane; the core compartment for its ERAD ligase activity. Reason: Core localization with direct experimental support. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005886 plasma membrane	EXP PMID:9533025 Cloning, expression and mapping of a novel RING-finger gene ...	KEEP AS NON CORE	Summary: Experimental localization to the plasma membrane reported in the original cloning study. A real but secondary localization. Reason: Documented in the cloning paper but the functionally dominant compartment is the ER membrane; plasma-membrane pool is peripheral to the core ERAD role. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt Predominantly located in the plasma membrane, with some localization occurring within cytoplasmic organelles
GO:0031966 mitochondrial membrane	EXP PMID:19285439 The ubiquitin ligase RNF5 regulates antiviral responses by m...	KEEP AS NON CORE	Summary: Experimental localization to the mitochondrial membrane, where RNF5 ubiquitinates STING1/MITA during antiviral responses. A real but secondary, immunity-related localization. Reason: Directly supported (PMID:19285439) but a secondary compartment for a context-specific immune role, not the core ER ERAD function. Supporting Evidence: PMID:19285439 virus-induced ubiquitination and degradation of MITA by RNF5 occurred at the mitochondria
GO:0061630 ubiquitin protein ligase activity	EXP PMID:19269966 Regulation of endoplasmic reticulum-associated degradation b...	ACCEPT	Summary: Experimental demonstration of RNF5 ubiquitin ligase activity (RING-dependent ubiquitination of JAMP). Core molecular function. Reason: Core molecular function with direct experimental support; WT but not RING-mutant RNF5 ubiquitinates substrate. Supporting Evidence: PMID:19269966 Ectopically expressed JAMP was efficiently ubiquitinated in vivo by WT but not the RING mutant form of RNF5
GO:0061630 ubiquitin protein ligase activity	EXP PMID:19285439 The ubiquitin ligase RNF5 regulates antiviral responses by m...	ACCEPT	Summary: Experimental demonstration of RNF5 ligase activity ubiquitinating STING1/MITA. Core molecular function. Reason: Core molecular function with direct experimental support. Supporting Evidence: PMID:19285439 RNF5 targeted MITA at Lys150 for ubiquitination and degradation after viral infection
GO:0061630 ubiquitin protein ligase activity	EXP PMID:23093945 Regulation of ATG4B stability by RNF5 limits basal levels of...	ACCEPT	Summary: Experimental demonstration of RNF5 ligase activity controlling ATG4B ubiquitination and stability. Core molecular function. Reason: Core molecular function with direct experimental support. Supporting Evidence: PMID:23093945 the membrane-associated E3 ligase RNF5 regulates basal levels of autophagy by controlling the stability of a select pool of the cysteine protease ATG4B
GO:0006511 ubiquitin-dependent protein catabolic process	IGI PMID:24019521 RNF185 is a novel E3 ligase of endoplasmic reticulum-associa...	ACCEPT	Summary: Genetic-interaction evidence (with RNF185) that RNF5 drives ubiquitin-dependent degradation of CFTR; redundant depletion blocks CFTR-deltaF508 turnover. Reason: Supported by the RNF5/RNF185 co-depletion experiments; consistent with the core ERAD/proteasomal degradation role. 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:0036503 ERAD pathway	IMP PMID:24019521 RNF185 is a novel E3 ligase of endoplasmic reticulum-associa...	ACCEPT	Summary: Mutant-phenotype evidence that RNF5 is required for ERAD of CFTR/CFTR-deltaF508. Core biological process. Reason: Core biological process with direct experimental (IMP) support from depletion experiments. Supporting Evidence: PMID:24019521 simultaneous depletion of RNF5 and RNF185 profoundly blocks CFTRΔF508 degradation
GO:0061630 ubiquitin protein ligase activity	TAS PMID:24019521 RNF185 is a novel E3 ligase of endoplasmic reticulum-associa...	ACCEPT	Summary: Author statement of RNF5 ubiquitin ligase activity in CFTR ERAD. Core molecular function. Reason: Correct core molecular function. Supporting Evidence: PMID:24019521 targeted to co-translational degradation by the E3 ligase RNF5/RMA1
GO:1904380 endoplasmic reticulum mannose trimming	TAS Reactome:R-HSA-901032	REMOVE	Summary: Reactome ERQC pathway annotation. RNF5 does not perform mannose trimming; this is pathway-adjacency over-annotation. Reason: RNF5 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/RNF5/RNF5-uniprot.txt Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
GO:0044877 protein-containing complex binding	IPI PMID:24019521 RNF185 is a novel E3 ligase of endoplasmic reticulum-associa...	KEEP AS NON CORE	Summary: RNF5 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:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8866542	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization within CFTR ERAD reactions. Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8866546	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (RNF5/RNF185 ubiquitinate misfolded CFTR). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8866551	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (CFTR binds ERAD machinery). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8866854	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (CFTR F508del translocation). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8866856	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (RNF5/RNF185 ubiquitinate CFTR F508del). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8866857	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (CFTR F508del binds ERAD machinery). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-8867288	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization in an ERAD ubiquitination reaction. Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-9931264	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (CD274/PD-L1 ERAD transport). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-9931298	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (ubiquitination of CD274 by ERAD complex). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
GO:0005789 endoplasmic reticulum membrane	TAS Reactome:R-HSA-9931313	ACCEPT	Summary: Reactome curation of RNF5 ER membrane localization (p-CD274 binds ERAD complex). Core compartment. Reason: Correct core localization; redundant with experimental evidence. Supporting Evidence: PMID:19285439 Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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:0030163 protein catabolic process	IMP PMID:19285439 The ubiquitin ligase RNF5 regulates antiviral responses by m...	KEEP AS NON CORE	Summary: Mutant-phenotype evidence that RNF5 drives degradation of a target protein (STING1/MITA). A parent of the more specific ubiquitin-dependent catabolic/ERAD process. Reason: Correct but generic relative to the specific ERAD and K48-ubiquitination annotations; here the catabolic target is the immune adaptor MITA. Supporting Evidence: PMID:19285439 RNF5 targeted MITA at Lys150 for ubiquitination and degradation after viral infection
GO:0070936 protein K48-linked ubiquitination	IDA PMID:19285439 The ubiquitin ligase RNF5 regulates antiviral responses by m...	ACCEPT	Summary: Direct evidence that RNF5 builds K48-linked polyubiquitin chains (degradative topology) on STING1/MITA. This degradative topology is also used on CFTR. Reason: Directly demonstrated K48-linked ubiquitination, the canonical degradative topology underlying RNF5's ERAD and MITA-degradation roles. Supporting Evidence: PMID:19269966 CFTR ubiquitination occurred at the canonical Lys-48 topology
GO:0004842 ubiquitin-protein transferase activity	IDA PMID:19269966 Regulation of endoplasmic reticulum-associated degradation b...	ACCEPT	Summary: Direct evidence 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 RING E3 ligase activity. Supporting Evidence: PMID:19269966 Ectopically expressed JAMP was efficiently ubiquitinated in vivo by WT but not the RING mutant form of RNF5
GO:0005515 protein binding	IPI PMID:19269966 Regulation of endoplasmic reticulum-associated degradation b...	KEEP AS NON CORE	Summary: Interaction with JKAMP/JAMP, a functionally important RNF5 substrate/partner at the ER membrane. Bare protein binding is uninformative. Reason: Records the real RNF5-JAMP interaction but bare protein binding is uninformative. Supporting Evidence: PMID:19269966 RNF5 associates with JAMP in the ER membrane
GO:0036503 ERAD pathway	IMP PMID:19269966 Regulation of endoplasmic reticulum-associated degradation b...	ACCEPT	Summary: Mutant-phenotype evidence linking RNF5 to ERAD, here via regulation of the ERAD adaptor JAMP. Core biological process. Reason: Core biological process; RNF5 modulates ERAD both by direct substrate ubiquitination and by K63-regulation of JAMP. Supporting Evidence: PMID:19269966 RNF5 is a ubiquitin ligase anchored to the ER membrane implicated in ERAD via ubiquitination of misfolded proteins
GO:0070534 protein K63-linked ubiquitination	IDA PMID:19269966 Regulation of endoplasmic reticulum-associated degradation b...	ACCEPT	Summary: Direct evidence that RNF5 builds non-degradative K63-linked polyubiquitin chains on JAMP (Ubc13/UBE2N-dependent), a regulatory rather than degradative modification. Reason: Directly demonstrated K63-linked ubiquitination, establishing RNF5's capacity for non-canonical regulatory ubiquitination distinct from K48 degradative chains. Supporting Evidence: PMID:19269966 These findings establish that RNF5 mediates Lys-63-based polyubiquitination of JAMP
GO:0008270 zinc ion binding	TAS PMID:9533025 Cloning, expression and mapping of a novel RING-finger gene ...	KEEP AS NON CORE	Summary: The C3HC4 RING domain coordinates zinc as a structural requirement for the ligase fold. A structural attribute supporting, not equal to, the ligase activity. Reason: Accurate structural feature of the RING domain (residues 27-68) but subsidiary to the informative ubiquitin ligase activity; not a standalone core function. Supporting Evidence: file:human/RNF5/RNF5-uniprot.txt RING-type

Core Functions

RING-type E3 ubiquitin ligase anchored in the ER membrane that ubiquitinates misfolded membrane-protein clients (e.g. CFTR and CFTR-deltaF508) with K48-linked chains to commit them to retrotranslocation and proteasomal degradation via the ERAD pathway, acting partly redundantly with RNF185.

Molecular Function:

ubiquitin protein ligase activity

Directly Involved In:

ERAD pathway

Cellular Locations:

endoplasmic reticulum membrane

Supporting Evidence:

PMID:16901789
an ER membrane-associated ubiquitin ligase complex containing the E3 RMA1, the E2 Ubc6e, and Derlin-1
PMID:24019521
identify RNF185 and RNF5 as a novel E3 ligase module that is central to the control of CFTR degradation

ER-membrane ubiquitin ligase that builds non-degradative K63-linked polyubiquitin chains on the ERAD adaptor JKAMP/JAMP (UBE2N/Ubc13-dependent), modulating its recruitment of proteasome and p97/VCP components and thereby regulating ERAD flux.

Molecular Function:

ubiquitin protein ligase activity

Directly Involved In:

protein K63-linked ubiquitination

Cellular Locations:

endoplasmic reticulum membrane

Supporting Evidence:

PMID:19269966
These findings establish that RNF5 mediates Lys-63-based polyubiquitination of JAMP

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:0000052

Gene Ontology annotation based on curation of immunofluorescence data

GO_REF:0000107

Automatic transfer of experimentally verified manual GO annotation data to orthologs using Ensembl Compara

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:12861019

RNF5, a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization.

RNF5 associates with the N-terminal domain of paxillin and mediates its ubiquitination, requiring intact RING and C-terminal domains, regulating cell motility.

PMID:14667819

Analysis of a high-throughput yeast two-hybrid system and its use to predict the function of intracellular proteins encoded within the human MHC class III region.

PMID:16901789

Sequential quality-control checkpoints triage misfolded cystic fibrosis transmembrane conductance regulator.

An ER membrane-associated ubiquitin ligase complex of E3 RMA1/RNF5, E2 Ubc6e and Derlin-1 recognizes folding defects in CFTR/CFTR-deltaF508 co-translationally and cooperates with cytosolic Hsc70/CHIP to promote proteasomal degradation.

PMID:19269966

Regulation of endoplasmic reticulum-associated degradation by RNF5-dependent ubiquitination of JNK-associated membrane protein (JAMP).

RNF5 builds Ubc13(UBE2N)-dependent K63-linked polyubiquitin chains on the ERAD adaptor JAMP at the ER membrane, reducing JAMP association with proteasome subunits and p97/VCP and thereby limiting ERAD; RNF5 builds canonical K48 chains on CFTR.

PMID:19285439

The ubiquitin ligase RNF5 regulates antiviral responses by mediating degradation of the adaptor protein MITA.

RNF5 interacts with STING1/MITA in a virus-infection-dependent manner and builds K48-linked chains on MITA at Lys150 for proteasomal degradation at mitochondria, negatively regulating type I interferon antiviral responses; RNF5 localizes at ER and mitochondria.

PMID:19549727

Analysis of the human E2 ubiquitin conjugating enzyme protein interaction network.

PMID:20152160

Crystal structure of UbcH5b~ubiquitin intermediate: insight into the formation of the self-assembled E2~Ub conjugates.

PMID:23093945

Regulation of ATG4B stability by RNF5 limits basal levels of autophagy and influences susceptibility to bacterial infection.

RNF5 regulates basal autophagy by controlling the stability of a membrane pool of the cysteine protease ATG4B, thereby limiting LC3/ATG8 processing required for autophagosome formation.

PMID:24019521

RNF185 is a novel E3 ligase of endoplasmic reticulum-associated degradation (ERAD) that targets cystic fibrosis transmembrane conductance regulator (CFTR).

RNF185 and RNF5 form a partly redundant E3 ligase module central to CFTR/CFTR-deltaF508 degradation; co-depletion profoundly blocks CFTR-deltaF508 degradation during and after translation.

PMID:25416956

A proteome-scale map of the human interactome network.

PMID:25759021

Regulation of glutamine carrier proteins by RNF5 determines breast cancer response to ER stress-inducing chemotherapies.

RNF5 regulates glutamine carrier proteins (SLC1A5, SLC38A2) under ER stress, influencing breast cancer response to chemotherapy.

PMID:26618866

∆F508 CFTR interactome remodelling promotes rescue of cystic fibrosis.

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:32814053

Interactome Mapping Provides a Network of Neurodegenerative Disease Proteins and Uncovers Widespread Protein Aggregation in Affected Brains.

PMID:33961781

Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.

PMID:36012204

Differential CFTR-Interactome Proximity Labeling Procedures Identify Enrichment in Multiple SLC Transporters.

PMID:9533025

Cloning, expression and mapping of a novel RING-finger gene (RNF5), a human homologue of a putative zinc-finger gene from Caenorhabditis elegans.

Original cloning of RNF5, a RING-finger gene in the MHC class III region; reported widespread expression and predominantly plasma-membrane localization with some cytoplasmic-organelle localization.

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:39098896

RING finger protein 5 protects against acute myocardial infarction by inhibiting ASK1.

RNF5 is downregulated in infarcted heart tissue; RNF5 knockout worsens and RNF5 overexpression ameliorates myocardial infarction injury in mouse and cardiomyocyte models, with protection attributed to inhibition of ASK1 (MAP3K5) activation.

Suggested Experiments

Experiment: Reconstitute ERAD ubiquitination of CFTR-deltaF508 in vitro with purified RNF5, RNF185, UBE2J1/Ubc6e and Derlin-1 to map lysine sites and ubiquitin-chain linkage on the substrate and to dissect RNF5/RNF185 redundancy.

Experiment: Generate RNF5 and RNF5/RNF185 double-knockout cells and perform quantitative ubiquitinome/proteome profiling under basal and ER-stress conditions to define the endogenous ERAD substrate repertoire and the degree of paralog redundancy.

Deep Research

Falcon

(RNF5-deep-research-falcon.md)

this with annotations you find in gene/protein databases, but these can be outdated or inaccurate. Falcon Edison Scientific Literature 20 citations 2 artifacts 2026-06-12T05:02:08.223244

Edison artifact artifact-00 (text/markdown)

## Context ID: pqac-00000017 The document does not contain a schematic labeled as Figure 1 that summarizes the pathway; Figure 1 in this paper focuses on charac (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 RNF5 (UniProt Q99942) functional annotation (2023–2024 emphasis)

0) Target verification (critical)

The requested target is human RNF5 (ring finger protein 5), UniProt Q99942, also known as RMA1. Recent CFTR/ERAD and innate-immunity literature consistently describes RNF5 as an ER-anchored/ER-embedded RING E3 ubiquitin ligase with C-terminal transmembrane segment(s), matching the UniProt-provided identity and domain architecture (RING finger with membrane anchor). (okiyoneda2024targetingubiquitinationmachinery pages 4-7, brusa2023innovativestrategytoward pages 9-11, kamada2024herc3facilitateserad pages 1-2)

1) Key concepts and definitions (current understanding)

1.1 Ubiquitination and E3 ligases

RNF5 functions as an E3 ubiquitin ligase, i.e., an enzyme that catalyzes transfer of ubiquitin from an E2 enzyme to substrate proteins, thereby modulating substrate fate (often proteasomal degradation depending on ubiquitin chain topology). In the contexts retrieved here, RNF5 is repeatedly linked to proteasome-directed ubiquitination, particularly in ER quality control and innate immune adaptor turnover. (ge2024rnf5inhibitingantiviral pages 4-5, okiyoneda2024targetingubiquitinationmachinery pages 4-7)

1.2 ER-associated degradation (ERAD) and ER quality control (ERQC)

ERAD is the principal pathway for disposing of misfolded or unassembled proteins in the endoplasmic reticulum. RNF5 is one of several ER membrane E3 ligases that recognize misfolded membrane proteins and initiate ubiquitination, targeting them for extraction and degradation by the proteasome. In CFTR biology, RNF5 is described as an ER-anchored E3 that detects folding/assembly defects at early biogenesis stages and contributes to degradation of misfolded CFTR variants such as F508del. (okiyoneda2024targetingubiquitinationmachinery pages 4-7, kamada2024herc3facilitateserad pages 1-2)

1.3 Innate immune adaptor regulation (cGAS–STING and RIG-I/MAVS)

In antiviral innate immunity, adaptor proteins such as STING (MITA) and MAVS (VISA) drive type I interferon signaling. RNF5 is described as a negative regulator of these pathways by mediating ubiquitination and degradation of key adaptors (e.g., STING, MAVS) and downstream signaling proteins (e.g., IRF3 via recruitment mechanisms). (ge2024rnf5inhibitingantiviral pages 5-7, ge2024rnf5inhibitingantiviral pages 4-5)

2) RNF5 molecular function, localization, and substrate specificity

2.1 Subcellular localization

Across multiple sources, RNF5 is described as ER-anchored/ER-embedded and membrane-associated, consistent with its principal roles in ERQC/ERAD and membrane-proximal innate signaling regulation. (okiyoneda2024targetingubiquitinationmachinery pages 4-7, kamada2024herc3facilitateserad pages 1-2)

2.2 RNF5 in CFTR ERAD: substrate recognition and pathway placement

CFTR-F508del is a misfolding-prone variant targeted for ERAD. RNF5 is described as recognizing folding/assembly defects in CFTR, including defects sensed at the N-terminus, and operating sequentially with CHIP (RNF5 early; CHIP later/post-translational) in monitoring CFTR folding status. (okiyoneda2024targetingubiquitinationmachinery pages 4-7)

Mechanistic placement supported by several sources:
- RNF5 contributes to ubiquitination of immature/core-glycosylated CFTR (Band B) rather than the mature plasma-membrane form (Band C), consistent with an early ER quality-control role. (riepe2024smallmoleculecorrectorsdivert pages 3-4)
- RNF5 was described as recognizing N-terminal regions/MSD1 and initiating ubiquitin chain formation that can be elongated by gp78/AMFR to facilitate ERAD. (kamada2024herc3facilitateserad pages 1-2)
- RNF5 shows redundancy/compensation with RNF185 (another ER membrane E3), which helps explain why single-gene perturbations can yield modest stabilization. (riepe2024smallmoleculecorrectorsdivert pages 3-4, okiyoneda2024targetingubiquitinationmachinery pages 11-13)

2.3 Innate immunity substrates: STING, MAVS, IRF3

A 2024 review focused on RNF5 in antiviral signaling summarizes that RNF5 promotes ubiquitination and degradation of STING and MAVS, thereby inhibiting antiviral innate immunity. (ge2024rnf5inhibitingantiviral pages 5-7, ge2024rnf5inhibitingantiviral pages 4-5)

Residue-level details reported (from sources cited within the 2024 MI study’s mechanistic background and consistent with the innate-immunity review):
- STING/MITA targeted at K150 (degradative ubiquitination context). (wan2024ringfingerprotein pages 12-13, ge2024rnf5inhibitingantiviral pages 4-5)
- MAVS/VISA ubiquitination at K362 and K461 with proteasome-dependent degradation. (wan2024ringfingerprotein pages 12-13, ge2024rnf5inhibitingantiviral pages 4-5)
- Activated IRF3 can be degraded in a complex where JMJD6 recruits RNF5, as summarized in the innate-immunity-focused review. (ge2024rnf5inhibitingantiviral pages 4-5)

RNF5 has also been connected to autophagy regulation by controlling the stability of autophagy proteins. In the context of RNF5 inhibition strategies in CF, RNF5 inhibition (inh-2 and optimized analogues) is reported to increase basal autophagy, consistent with prior descriptions that RNF5 can promote ATG4B degradation and that RNF5 inhibitors interfere with this effect. (brusa2023innovativestrategytoward pages 9-11, brusa2023innovativestrategytoward pages 14-15)

2.5 Cardiovascular signaling (ASK1)

A 2024 mouse/cardiomyocyte study proposes a cardioprotective role for RNF5, reporting that RNF5 downregulation occurs in infarcted heart tissue and that RNF5 knockout worsens myocardial infarction phenotypes, while RNF5 overexpression is protective; mechanistically, protection is attributed to inhibition of ASK1 activation (phosphorylation), though the directness of RNF5’s E3 action on ASK1 is not fully resolved in the provided excerpt. (wan2024ringfingerprotein pages 12-13)

3) Recent developments (prioritizing 2023–2024)

3.1 Genome-wide CRISPR screens redefine RNF5’s role as important but partially redundant in CFTR-F508del ERAD (2024)

A 2024 Molecular Biology of the Cell study performed genome-wide CRISPR/Cas9 screens to identify machinery driving CFTR-F508del ERAD.
- Scale: 20,528 genes screened; 207 high-confidence hits at FDR < 1%. (riepe2024smallmoleculecorrectorsdivert pages 3-4)
- Key result: RNF5 was the top E3 ligase hit, but RNF5 knockout only modestly reduced CFTR-F508del degradation, supporting robustness and redundancy. (riepe2024smallmoleculecorrectorsdivert pages 3-4, riepe2024smallmoleculecorrectorsdivert pages 1-2)
- Redundancy: secondary screens in an RNF5 knockout background identified RNF185 as a redundant ligase. (riepe2024smallmoleculecorrectorsdivert pages 3-4, okiyoneda2024targetingubiquitinationmachinery pages 11-13)
- Quantitative kinetics: CFTR-F508del reporter showed a degradation half-life of about 34 minutes in the reported system. (riepe2024smallmoleculecorrectorsdivert pages 3-4)

A schematic (Figure 4A) from this work summarizes sequential folding states and proposes that small-molecule correctors stabilize folding states that are not RNF5 substrates. (riepe2024smallmoleculecorrectorsdivert media 192d3a11)

3.2 Medicinal chemistry progress: thiadiazole RNF5 inhibitors that enhance mutant CFTR rescue (2023)

A 2023 Journal of Medicinal Chemistry study advanced RNF5 inhibition as a CF strategy.
- Inhibitor scaffold: inh-2 (1,2,4-thiadiazol-5-ylidene) described as a first-in-class RNF5 inhibitor, with SAR leading to improved analogues including compound/analogue 16. (brusa2023innovativestrategytoward pages 14-15, brusa2023innovativestrategytoward pages 1-3)
- On-target validation: analogue 16 improved CFTR rescue in control (NT siRNA) cells but not in RNF5 siRNA cells, consistent with RNF5-dependent activity. (brusa2023innovativestrategytoward pages 11-14)
- Experimental concentrations (useful for real-world reproducibility): compounds commonly tested at 5 μM, with CFTR correctors used at VX-809 1–3 μM, VX-445 3 μM, and VX-661 + VX-445 10 μM + 3 μM; MG-132 used at 10 μM for ubiquitination assays. (brusa2023innovativestrategytoward pages 11-14)
- Autophagy link: analogue 16 increased basal autophagy, consistent with RNF5 inhibition phenotypes. (brusa2023innovativestrategytoward pages 11-14, brusa2023innovativestrategytoward pages 14-15)

3.3 Updated CF translational assessment: no ubiquitination-pathway drugs in CF clinical trials (2024 review)

A 2024 review focused on targeting ubiquitination machinery in CF notes that, despite long-standing interest, no compounds targeting ubiquitination have entered clinical trials for CF (at least as of June 2024 publication). (okiyoneda2024targetingubiquitinationmachinery pages 4-7)

The same review summarizes:
- RNF5 as an ER-anchored E3 that recognizes CFTR folding defects, pairs with E2 enzymes (e.g., UBE2J1), and acts sequentially with CHIP. (okiyoneda2024targetingubiquitinationmachinery pages 4-7)
- Preclinical results in which in vivo suppression of RNF5 in F508del-CFTR transgenic mice improved intestinal malabsorption and increased CFTR activity in intestinal epithelial cells. (okiyoneda2024targetingubiquitinationmachinery pages 4-7)

3.4 RNF5 as an innate-immunity node and potential host-directed antiviral target (2024 review)

A 2024 Frontiers in Immunology review describes RNF5 as a key modulator of antiviral signaling and virus life cycle through ubiquitination of host innate-immune proteins (STING, MAVS, IRF3) and also viral proteins (notably in SARS-CoV-2 and KSHV contexts). (ge2024rnf5inhibitingantiviral pages 5-7, ge2024rnf5inhibitingantiviral pages 7-9)

4) Current applications and real-world implementations

4.1 Cystic fibrosis: modulating RNF5-mediated ubiquitination to rescue CFTR

The most concrete application in the retrieved corpus is RNF5 inhibition to stabilize immature F508del-CFTR and improve response to CFTR correctors.
- Real-world relevance: Over 80% of people with CF carry the F508del mutation, making the pathway broadly relevant to CF therapeutics. (riepe2024smallmoleculecorrectorsdivert pages 1-2)
- Implementation status: RNF5 inhibitors (inh-2, analogue 16) have been tested in airway epithelial models and in combination with clinically used correctors, but translation remains preclinical, and reviews emphasize lack of clinical trials targeting ubiquitination in CF. (okiyoneda2024targetingubiquitinationmachinery pages 4-7, brusa2023innovativestrategytoward pages 11-14)

4.2 Host-directed antiviral strategies

RNF5 modulation (both inhibition and activation) is discussed as a strategy to shift antiviral signaling and/or viral replication steps, but this is currently largely mechanistic/preclinical in the provided corpus. (ge2024rnf5inhibitingantiviral pages 5-7, ge2024rnf5inhibitingantiviral pages 7-9)

4.3 Cardiovascular disease models

RNF5 has been proposed as a cardioprotective regulator in myocardial infarction via the ASK1 pathway, based on genetic manipulation in mice and cardiomyocyte models; translation to therapy is not yet demonstrated in the provided evidence. (wan2024ringfingerprotein pages 12-13)

5) Expert opinions and analysis (authoritative perspectives)

5.1 CF field perspective: redundancy and risk-benefit constraints

The 2024 CF ubiquitination review emphasizes both the appeal and difficulty of targeting ubiquitination: blocking ubiquitination to stabilize mutant CFTR has been considered for a long time, but progress has been slow and clinical translation has not yet occurred. It also highlights that approved corrector combinations (e.g., Trikafta/Kaftrio) demonstrate the effectiveness of pharmacological chaperone approaches that do not broadly interfere with cell signaling. (okiyoneda2024targetingubiquitinationmachinery pages 4-7)

5.2 Systems view from CRISPR genetics: RNF5 is a major node but not sufficient alone

The 2024 CRISPR screen study shows RNF5 is the top E3 hit for CFTR-F508del ERAD, but RNF5 loss produces only modest stabilization, consistent with redundant E3 ligase networks (RNF185 and others). This has practical implications: RNF5 inhibitors may need combination approaches or context-dependent use to yield clinically meaningful stabilization. (riepe2024smallmoleculecorrectorsdivert pages 3-4, okiyoneda2024targetingubiquitinationmachinery pages 11-13)

5.3 Innate immunity perspective: RNF5 as a negative regulator of STING/MAVS

The 2024 RNF5 antiviral-immunity review frames RNF5 as a “novel means of modulating antiviral immunity,” emphasizing both host control mechanisms and viral exploitation of RNF5. This positions RNF5 as a plausible therapeutic lever, but also highlights pathway complexity and context-dependent outcomes. (ge2024rnf5inhibitingantiviral pages 5-7, ge2024rnf5inhibitingantiviral pages 7-9)

6) Relevant statistics and data points (recent studies)

CF genetics: F508del is present in >80% of people with CF (reported in 2024 MBoC paper) and affects ~80% of CF patients worldwide (reported in 2023 J Med Chem paper). (riepe2024smallmoleculecorrectorsdivert pages 1-2, brusa2023innovativestrategytoward pages 1-3)
CRISPR screen scale (2024): 20,528 genes screened; 207 high-confidence hits at FDR < 1%. (riepe2024smallmoleculecorrectorsdivert pages 3-4)
CFTR-F508del ERAD kinetics: majority of reporter degraded with half-life ~34 min in the described system. (riepe2024smallmoleculecorrectorsdivert pages 3-4)
Drug/chemical biology concentrations used (2023): RNF5 inhibitors tested typically at 5 μM; CFTR correctors VX-809 1–3 μM, VX-445 3 μM, VX-661 + VX-445 10 μM + 3 μM; MG-132 10 μM. (brusa2023innovativestrategytoward pages 11-14)
Open Targets disease association evidence (database aggregation): RNF5 is associated with multiple diseases including cystic fibrosis and neoplasms; example association scores shown in the retrieved Open Targets output include cystic fibrosis 0.4753, neoplasm 0.1277, glioblastoma multiforme 0.1098, acute lymphoblastic leukemia 0.1049, and experimental autoimmune encephalomyelitis 0.1039 (note: these are Open Targets model scores, not effect sizes from a single experiment). (OpenTargets Search: -RNF5)

7) Mechanistic summary figure

The following image provides a mechanistic synthesis of CFTR-F508del folding states and the proposed point at which RNF5-mediated ubiquitination targets early folding intermediates, while correctors stabilize RNF5-resistant states. (riepe2024smallmoleculecorrectorsdivert media 192d3a11)

8) Summary and functional annotation statement

Human RNF5 (UniProt Q99942) is best supported as an ER-embedded RING E3 ubiquitin ligase that (i) participates in ER quality control/ERAD of misfolded membrane proteins—most prominently CFTR-F508del—by acting at early biogenesis stages on immature ER-resident intermediates, in collaboration with specific E2 enzymes and partially redundant ER E3 ligases; and (ii) regulates innate immunity by promoting ubiquitination-dependent degradation of signaling adaptors such as STING and MAVS, thereby dampening antiviral interferon signaling; RNF5 also interfaces with autophagy regulation (e.g., ATG4B stability) and has emerging disease-model roles (e.g., ASK1 signaling in myocardial infarction). Recent 2023–2024 work positions RNF5 as an attractive but challenging therapeutic target because of network redundancy and the current gap between preclinical modulators (e.g., thiadiazole inhibitors) and clinical translation, particularly in cystic fibrosis. (ge2024rnf5inhibitingantiviral pages 5-7, okiyoneda2024targetingubiquitinationmachinery pages 4-7, riepe2024smallmoleculecorrectorsdivert pages 3-4, brusa2023innovativestrategytoward pages 11-14)

Appendix: compact evidence map

Biological context/pathway	RNF5 molecular action	Key substrates/partners and specific lysines/linkages when available	Subcellular localization context	Recent 2023–2024 findings/applications with quantitative details	Primary supporting source
ERAD / proteostasis	RING E3 ubiquitin ligase; ubiquitinates immature membrane proteins for proteasomal ERAD	CFTR N-terminus/MSD1 recognized; partners include UBE2J1, UBE2D3, RNF185, gp78/AMFR, HERC3; acts mainly on immature CFTR Band B (okiyoneda2024targetingubiquitinationmachinery pages 4-7, riepe2024smallmoleculecorrectorsdivert pages 3-4, okiyoneda2024targetingubiquitinationmachinery pages 11-13, kamada2024herc3facilitateserad pages 1-2)	ER-anchored / ER-embedded membrane ligase with C-terminal TM region(s) (okiyoneda2024targetingubiquitinationmachinery pages 4-7, brusa2023innovativestrategytoward pages 9-11, kamada2024herc3facilitateserad pages 1-2)	Genome-wide CRISPR screen assayed 20,528 genes and found 207 high-confidence hits (FDR <1%); RNF5 was top E3 hit, UBE2D3 top E2; RNF5 KO had only modest effect, supporting pathway redundancy (riepe2024smallmoleculecorrectorsdivert pages 3-4)	Small-molecule correctors divert CFTR-F508del from ERAD by stabilizing sequential folding states (2024), https://doi.org/10.1091/mbc.e23-08-0336
CFTR-F508del	Promotes ubiquitination and degradation of misfolded F508del-CFTR; early triage E3 in sequential ERQC	F508del-CFTR; sequential action with CHIP; RNF185 can compensate; correctors stabilize RNF5-resistant folding states (okiyoneda2024targetingubiquitinationmachinery pages 4-7, riepe2024smallmoleculecorrectorsdivert pages 3-4, okiyoneda2024targetingubiquitinationmachinery pages 11-13, riepe2024smallmoleculecorrectorsdivert media 192d3a11)	ER membrane / ERQC on nascent or immature CFTR before plasma-membrane maturation (okiyoneda2024targetingubiquitinationmachinery pages 4-7, riepe2024smallmoleculecorrectorsdivert pages 3-4, kamada2024herc3facilitateserad pages 1-2, riepe2024smallmoleculecorrectorsdivert media 192d3a11)	F508del occurs in >80% of people with CF / affects ~80% of CF patients worldwide; RNF5 suppression improved intestinal malabsorption and CFTR activity in F508del-transgenic mice; no RNF5-targeting compounds reported in clinical trials in the provided context (okiyoneda2024targetingubiquitinationmachinery pages 4-7, riepe2024smallmoleculecorrectorsdivert pages 1-2, brusa2023innovativestrategytoward pages 1-3)	Targeting ubiquitination machinery in cystic fibrosis: Where do we stand? (2024), https://doi.org/10.1007/s00018-024-05295-z
CFTR-F508del therapeutic targeting	Pharmacologic RNF5 inhibition / degradation to reduce CFTR ubiquitination	inh-2 (first-in-class RNF5 inhibitor), analogue 16, FX12; analogue 16 effect lost with RNF5 siRNA, supporting on-target action (brusa2023innovativestrategytoward pages 11-14, brusa2023innovativestrategytoward pages 14-15, brusa2023innovativestrategytoward pages 1-3)	ER-associated RNF5 targeted indirectly by small molecules in airway epithelial cell models (brusa2023innovativestrategytoward pages 11-14, brusa2023innovativestrategytoward pages 14-15)	Compound testing commonly at 5 μM; VX-809 1–3 μM, VX-445 3 μM, VX-661 + VX-445 10 μM + 3 μM, MG-132 10 μM; analogue 16 was among the most promising compounds (11, 16, 21) and improved rescue with ELX/TEZ/IVA in CFBE41o− cells; FX12 active in BHK cells but not differentiated primary airway epithelia (brusa2023innovativestrategytoward pages 11-14, brusa2023innovativestrategytoward pages 14-15, okiyoneda2024targetingubiquitinationmachinery pages 4-7)	Innovative Strategy toward Mutant CFTR Rescue in Cystic Fibrosis: Design and Synthesis of Thiadiazole Inhibitors of the E3 Ligase RNF5 (2023), https://doi.org/10.1021/acs.jmedchem.3c00608
Innate immunity: cGAS-STING / MAVS / IRF3	E3 ligase that mainly promotes degradative ubiquitination of antiviral signaling adaptors; K48 linked for STING/MAVS in mammalian studies summarized in review	STING/MITA at K150 (K48-linked); MAVS/VISA at K362 and K461; activated IRF3 degraded after recruitment by JMJD6; viral proteins can hijack RNF5 and alter linkage usage (K27/K29/K63 also discussed in viral contexts) (wan2024ringfingerprotein pages 12-13, ge2024rnf5inhibitingantiviral pages 9-10, ge2024rnf5inhibitingantiviral pages 4-5)	ER-associated; MAVS regulation occurs in mitochondria-associated antiviral signaling context; STING at ER/ERGIC-associated innate immune membranes (ge2024rnf5inhibitingantiviral pages 9-10, ge2024rnf5inhibitingantiviral pages 4-5)	2024 review emphasizes RNF5 as a negative regulator of antiviral innate immunity and a possible host-directed target; no clinical-stage RNF5 immune modulators reported in the provided context (ge2024rnf5inhibitingantiviral pages 5-7, ge2024rnf5inhibitingantiviral pages 9-10, ge2024rnf5inhibitingantiviral pages 4-5)	RNF5: inhibiting antiviral immunity and shaping virus life cycle (2024), https://doi.org/10.3389/fimmu.2023.1324516
Autophagy	Regulates autophagy by targeting autophagy machinery components for degradation	ATG4B is a reported RNF5 substrate; inh-2 and analogue 16 increase basal autophagy consistent with blocking RNF5-mediated turnover; PTGDR2 can compete with ATG4B for RNF5 binding in gastric CSC context (brusa2023innovativestrategytoward pages 11-14, brusa2023innovativestrategytoward pages 9-11, brusa2023innovativestrategytoward pages 14-15)	ER-associated RNF5 with effects on cytosolic autophagy regulators (brusa2023innovativestrategytoward pages 9-11, brusa2023innovativestrategytoward pages 14-15)	Autophagy vacuoles increased with inh-2 and compounds 11/16/21; torin-1 used at 20 nM, SAR-405 at 2 μM in functional autophagy assays (brusa2023innovativestrategytoward pages 11-14)	Innovative Strategy toward Mutant CFTR Rescue in Cystic Fibrosis: Design and Synthesis of Thiadiazole Inhibitors of the E3 Ligase RNF5 (2023), https://doi.org/10.1021/acs.jmedchem.3c00608
Cardiovascular	Protective E3-ligase-associated role in stress signaling; inhibits ASK1 pathway in MI model	ASK1 pathway implicated; whether direct ubiquitination of ASK1 is established was unresolved in provided context (wan2024ringfingerprotein pages 12-13)	Cardiac cells/tissue; RNF5 downregulated in infarcted mouse heart and OGD-treated cardiomyocytes (wan2024ringfingerprotein pages 12-13)	2024 mouse/cell study: RNF5 knockout worsened myocardial infarction-associated dysfunction, inflammation, and apoptosis, while overexpression was protective; quantitative effect sizes not provided in context (wan2024ringfingerprotein pages 12-13)	RING finger protein 5 protects against acute myocardial infarction by inhibiting ASK1 (2024), https://doi.org/10.1186/s12872-024-04070-z
Oncology / other	Context-dependent E3 ligase affecting tumor signaling, proteostasis, and therapeutic response	EphA2/Eph receptors, paxillin, ATG4B; Open Targets links RNF5 to neoplasm, glioblastoma, ALL, cystic fibrosis, and EAE based on supporting literature (ge2024rnf5inhibitingantiviral pages 7-9, ge2024rnf5inhibitingantiviral pages 9-10, OpenTargets Search: -RNF5)	ER-associated ligase with downstream effects on signaling, migration, and transcriptional programs (ge2024rnf5inhibitingantiviral pages 7-9, ge2024rnf5inhibitingantiviral pages 9-10)	Open Targets evidence scores in provided context: cystic fibrosis association 0.4753, neoplasm 0.1277, glioblastoma 0.1098, ALL 0.1049, EAE 0.1039; preclinical therapeutic interest noted in AML, KSHV/PEL, and breast-cancer signaling, but no clinical trials reported here (OpenTargets Search: -RNF5, ge2024rnf5inhibitingantiviral pages 9-10)	Open Targets disease-target associations for RNF5 (accessed via provided context) (OpenTargets Search: -RNF5)

Table: This table summarizes evidence-based functional annotation for human RNF5 (UniProt Q99942), organized by pathway context, molecular action, substrates, localization, and recent translational findings. It is useful as a compact reference for RNF5’s validated roles in ERAD, CFTR quality control, innate immunity, autophagy, cardiovascular biology, and disease associations.

References

(okiyoneda2024targetingubiquitinationmachinery pages 4-7): Tsukasa Okiyoneda, Christian Borgo, Valentina Bosello Travain, Nicoletta Pedemonte, and Mauro Salvi. Targeting ubiquitination machinery in cystic fibrosis: where do we stand? Cellular and Molecular Life Sciences: CMLS, Jun 2024. URL: https://doi.org/10.1007/s00018-024-05295-z, doi:10.1007/s00018-024-05295-z. This article has 6 citations.
(brusa2023innovativestrategytoward pages 9-11): Irene Brusa, Elvira Sondo, Emanuela Pesce, Valeria Tomati, Dario Gioia, Federico Falchi, Beatrice Balboni, Jose Antonio Ortega Martínez, Marina Veronesi, Elisa Romeo, Natasha Margaroli, Maurizio Recanatini, Stefania Girotto, Nicoletta Pedemonte, Marinella Roberti, and Andrea Cavalli. Innovative strategy toward mutant cftr rescue in cystic fibrosis: design and synthesis of thiadiazole inhibitors of the e3 ligase rnf5. Journal of Medicinal Chemistry, 66:9797-9822, Jul 2023. URL: https://doi.org/10.1021/acs.jmedchem.3c00608, doi:10.1021/acs.jmedchem.3c00608. This article has 14 citations and is from a highest quality peer-reviewed journal.
(kamada2024herc3facilitateserad pages 1-2): Yuka Kamada, Yuko Ohnishi, Chikako Nakashima, Aika Fujii, Mana Terakawa, Ikuto Hamano, Uta Nakayamada, Saori Katoh, Noriaki Hirata, Hazuki Tateishi, Ryosuke Fukuda, Hirotaka Takahashi, Gergely L. Lukacs, and Tsukasa Okiyoneda. Herc3 facilitates erad of select membrane proteins by recognizing membrane-spanning domains. The Journal of Cell Biology, May 2024. URL: https://doi.org/10.1083/jcb.202308003, doi:10.1083/jcb.202308003. This article has 13 citations.
(ge2024rnf5inhibitingantiviral pages 4-5): Junyi Ge and Leiliang Zhang. Rnf5: inhibiting antiviral immunity and shaping virus life cycle. Frontiers in Immunology, Jan 2024. URL: https://doi.org/10.3389/fimmu.2023.1324516, doi:10.3389/fimmu.2023.1324516. This article has 9 citations and is from a peer-reviewed journal.
(ge2024rnf5inhibitingantiviral pages 5-7): Junyi Ge and Leiliang Zhang. Rnf5: inhibiting antiviral immunity and shaping virus life cycle. Frontiers in Immunology, Jan 2024. URL: https://doi.org/10.3389/fimmu.2023.1324516, doi:10.3389/fimmu.2023.1324516. This article has 9 citations and is from a peer-reviewed journal.
(riepe2024smallmoleculecorrectorsdivert pages 3-4): 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.
(okiyoneda2024targetingubiquitinationmachinery pages 11-13): Tsukasa Okiyoneda, Christian Borgo, Valentina Bosello Travain, Nicoletta Pedemonte, and Mauro Salvi. Targeting ubiquitination machinery in cystic fibrosis: where do we stand? Cellular and Molecular Life Sciences: CMLS, Jun 2024. URL: https://doi.org/10.1007/s00018-024-05295-z, doi:10.1007/s00018-024-05295-z. This article has 6 citations.
(wan2024ringfingerprotein pages 12-13): Hong Wan, Jianqing Zhang, Zhen Liu, Bizhen Dong, Zhangqian Tao, Guanglin Wang, and Chihua Wang. Ring finger protein 5 protects against acute myocardial infarction by inhibiting ask1. BMC Cardiovascular Disorders, Aug 2024. URL: https://doi.org/10.1186/s12872-024-04070-z, doi:10.1186/s12872-024-04070-z. This article has 4 citations and is from a peer-reviewed journal.
(brusa2023innovativestrategytoward pages 14-15): Irene Brusa, Elvira Sondo, Emanuela Pesce, Valeria Tomati, Dario Gioia, Federico Falchi, Beatrice Balboni, Jose Antonio Ortega Martínez, Marina Veronesi, Elisa Romeo, Natasha Margaroli, Maurizio Recanatini, Stefania Girotto, Nicoletta Pedemonte, Marinella Roberti, and Andrea Cavalli. Innovative strategy toward mutant cftr rescue in cystic fibrosis: design and synthesis of thiadiazole inhibitors of the e3 ligase rnf5. Journal of Medicinal Chemistry, 66:9797-9822, Jul 2023. URL: https://doi.org/10.1021/acs.jmedchem.3c00608, doi:10.1021/acs.jmedchem.3c00608. This article has 14 citations and is from a highest quality peer-reviewed journal.
(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.
(riepe2024smallmoleculecorrectorsdivert media 192d3a11): 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.
(brusa2023innovativestrategytoward pages 1-3): Irene Brusa, Elvira Sondo, Emanuela Pesce, Valeria Tomati, Dario Gioia, Federico Falchi, Beatrice Balboni, Jose Antonio Ortega Martínez, Marina Veronesi, Elisa Romeo, Natasha Margaroli, Maurizio Recanatini, Stefania Girotto, Nicoletta Pedemonte, Marinella Roberti, and Andrea Cavalli. Innovative strategy toward mutant cftr rescue in cystic fibrosis: design and synthesis of thiadiazole inhibitors of the e3 ligase rnf5. Journal of Medicinal Chemistry, 66:9797-9822, Jul 2023. URL: https://doi.org/10.1021/acs.jmedchem.3c00608, doi:10.1021/acs.jmedchem.3c00608. This article has 14 citations and is from a highest quality peer-reviewed journal.
(brusa2023innovativestrategytoward pages 11-14): Irene Brusa, Elvira Sondo, Emanuela Pesce, Valeria Tomati, Dario Gioia, Federico Falchi, Beatrice Balboni, Jose Antonio Ortega Martínez, Marina Veronesi, Elisa Romeo, Natasha Margaroli, Maurizio Recanatini, Stefania Girotto, Nicoletta Pedemonte, Marinella Roberti, and Andrea Cavalli. Innovative strategy toward mutant cftr rescue in cystic fibrosis: design and synthesis of thiadiazole inhibitors of the e3 ligase rnf5. Journal of Medicinal Chemistry, 66:9797-9822, Jul 2023. URL: https://doi.org/10.1021/acs.jmedchem.3c00608, doi:10.1021/acs.jmedchem.3c00608. This article has 14 citations and is from a highest quality peer-reviewed journal.
(ge2024rnf5inhibitingantiviral pages 7-9): Junyi Ge and Leiliang Zhang. Rnf5: inhibiting antiviral immunity and shaping virus life cycle. Frontiers in Immunology, Jan 2024. URL: https://doi.org/10.3389/fimmu.2023.1324516, doi:10.3389/fimmu.2023.1324516. This article has 9 citations and is from a peer-reviewed journal.
(OpenTargets Search: -RNF5): Open Targets Query (-RNF5, 38 results). Buniello, A. et al. (2025). Open Targets Platform: facilitating therapeutic hypotheses building in drug discovery. Nucleic Acids Research.
(ge2024rnf5inhibitingantiviral pages 9-10): Junyi Ge and Leiliang Zhang. Rnf5: inhibiting antiviral immunity and shaping virus life cycle. Frontiers in Immunology, Jan 2024. URL: https://doi.org/10.3389/fimmu.2023.1324516, doi:10.3389/fimmu.2023.1324516. This article has 9 citations and is from a peer-reviewed journal.

Artifacts

Edison artifact artifact-00

Citations

okiyoneda2024targetingubiquitinationmachinery pages 4-7
riepe2024smallmoleculecorrectorsdivert pages 3-4
wan2024ringfingerprotein pages 12-13
brusa2023innovativestrategytoward pages 11-14
riepe2024smallmoleculecorrectorsdivert pages 1-2
brusa2023innovativestrategytoward pages 9-11
okiyoneda2024targetingubiquitinationmachinery pages 11-13
brusa2023innovativestrategytoward pages 14-15
brusa2023innovativestrategytoward pages 1-3
https://doi.org/10.1091/mbc.e23-08-0336
https://doi.org/10.1007/s00018-024-05295-z
https://doi.org/10.1021/acs.jmedchem.3c00608
https://doi.org/10.3389/fimmu.2023.1324516
https://doi.org/10.1186/s12872-024-04070-z
https://doi.org/10.1007/s00018-024-05295-z,
https://doi.org/10.1021/acs.jmedchem.3c00608,
https://doi.org/10.1083/jcb.202308003,
https://doi.org/10.3389/fimmu.2023.1324516,
https://doi.org/10.1091/mbc.e23-08-0336,
https://doi.org/10.1186/s12872-024-04070-z,

📚 Additional Documentation

Notes

(RNF5-notes.md)

RNF5 (RMA1) review notes

UniProt: Q99942 (RNF5_HUMAN), 180 aa, gene RNF5 / synonyms RMA1, NG2, G16. HGNC:10068. Chromosome 6 MHC class III region.

Core identity

RNF5 is a membrane-anchored RING-type E3 ubiquitin ligase (EC 2.3.2.27). It has a C3HC4 RING domain (ZN_FING 27-68; catalytic Cys-42, mutation C42S abolishes ligase activity) and two C-terminal transmembrane helices (118-138, 160-180) that anchor it to the ER membrane (tail-anchored / multi-pass).

[file:human/RNF5/RNF5-uniprot.txt "Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins"]
[file:human/RNF5/RNF5-uniprot.txt "MUTAGEN 42 ... C->S: Loss of E3 ubiquitin-protein ligase activity"]

Primary role: ERAD of misfolded membrane proteins (incl. CFTR/CFTRdeltaF508)

RNF5/RMA1 is one of the founding mammalian ER-associated E3 ligases for ERAD. It primes misfolded CFTR/CFTRdeltaF508 for degradation co-translationally, acting in an ER-membrane complex with the E2 Ubc6e and Derlin-1, upstream of/parallel to the cytosolic Hsc70/CHIP branch.

PMID:16901789
PMID:24019521

Functional redundancy with RNF185

RNF5 and its paralog RNF185 form a partly redundant ERAD E3 module for CFTR; simultaneous depletion profoundly blocks CFTRdeltaF508 degradation. RNF5 interacts physically with RNF185 (IntAct).

PMID:24019521
PMID:24019521

E2 partners / ubiquitin chain topologies

RNF5 works with UBE2D (UbcH5) family E2s and can use Ubc13 (UBE2N) for K63 chains. It builds K48 chains on CFTR (degradative) and K63 chains on JAMP/JKAMP (non-degradative, regulatory).

PMID:19269966
PMID:19269966
[file:human/RNF5/RNF5-uniprot.txt "Mediates the 'Lys-63'-linked polyubiquitination of JKAMP thereby regulating JKAMP function by decreasing its association with components of the proteasome and ERAD; the ubiquitination appears to involve E2 ubiquitin-conjugating enzyme UBE2N"]

Secondary / non-core roles (pleiotropic)

Antiviral immunity: K48-linked ubiquitination and degradation of STING1/MITA at Lys-150 at mitochondria, negatively regulating type I IFN responses. Note this is at mitochondria, not ER, and is a regulatory immune role.
PMID:19285439
PMID:19285439
Autophagy: regulates basal autophagy by controlling stability of membrane-associated ATG4B, limiting LC3 processing.
PMID:23093945
Cell motility: non-canonical (Ubc13/K63) ubiquitination of paxillin (PXN), altering its localization.
PMID:12861019
Breast cancer / glutamine carrier (SLC1A5, SLC38A2) regulation under ER stress (PMID:25759021).

Localization

UniProt records Cell membrane (plasma membrane, PMID:9533025 early study), Mitochondrion membrane (PMID:19285439), and ER membrane (PMID:19285439). The functionally dominant ERAD location is the ER membrane. Plasma membrane and mitochondrial membrane localizations are real but reflect the original cloning paper and the antiviral/MITA study respectively. ER membrane is the core compartment for the ligase's ERAD function.

[file:human/RNF5/RNF5-uniprot.txt "Endoplasmic reticulum membrane {ECO:0000269|PubMed:19285439}; Multi-pass membrane protein"]

Annotation review decisions (summary)

ubiquitin protein ligase activity (GO:0061630): CORE, ACCEPT (IBA/IEA/TAS/EXP all supported; EC 2.3.2.27).
ERAD pathway (GO:0036503): CORE, ACCEPT (IBA, IMP PMID:24019521 & PMID:19269966).
ER membrane (GO:0005789): CORE location, ACCEPT.
zinc ion binding (GO:0008270): KEEP_AS_NON_CORE (structural RING feature).
protein binding (GO:0005515, many IPI): KEEP_AS_NON_CORE (uninformative; includes meaningful E2 and CFTR/Derlin partners but bare term not core).
ubiquitin-like protein conjugating enzyme binding (GO:0044390): ACCEPT non-core / KEEP_AS_NON_CORE — reflects E2 (UBE2D/UBE2N) binding, informative-ish but ancillary to ligase activity.
K48-linked (GO:0070936) and K63-linked (GO:0070534) ubiquitination: ACCEPT (IDA, both topologies demonstrated).
protein-containing complex binding (GO:0044877): KEEP_AS_NON_CORE.
plasma membrane (GO:0005886), mitochondrial membrane (GO:0031966), membrane (GO:0016020): KEEP_AS_NON_CORE (real but secondary; ER membrane is core).
transmembrane transport (GO:0055085, Reactome TAS): MARK_AS_OVER_ANNOTATED — RNF5 is not a transporter; this is pathway-context (ABC transport / CFTR) bleed-through, not a direct RNF5 function.
endoplasmic reticulum mannose trimming (GO:1904380, IEA ARBA / Reactome TAS): MARK_AS_OVER_ANNOTATED — RNF5 does not trim mannose; this is pathway-adjacency over-annotation.
ER quality control compartment (GO:0044322, IEA inter-ontology link): KEEP_AS_NON_CORE / ACCEPT — plausible localization from ERAD inference.
identical protein binding (GO:0042802): KEEP_AS_NON_CORE (RNF5 self-interaction in interactome screens).

Falcon deep-research findings (incorporated 2026-06)

Reviewed RNF5-deep-research-falcon.md (Edison/Falcon, 2026-06-12). Most content (ERAD/CFTR-F508del, STING/MAVS antiviral negative regulation, ATG4B/autophagy, RNF5/RNF185 redundancy) is already captured in this review. Genuinely new items:

NEW cardioprotective role via ASK1: RNF5 is downregulated in infarcted heart; RNF5 KO worsens and overexpression ameliorates myocardial infarction, attributed to inhibition of ASK1 (MAP3K5) activation. PMID:39098896 (DOI 10.1186/s12872-024-04070-z). Added to references (relevance MEDIUM); directness of RNF5 ubiquitination of ASK1 not established in the abstract, and full text is not cached, so no GO annotation or supporting_text was added.
2024 genome-wide CRISPR screen (Riepe et al., MBoC, DOI 10.1091/mbc.e23-08-0336) confirms RNF5 is the top E3 hit but only modest single-KO effect on CFTR-F508del ERAD, with RNF185 as the redundant ligase and UBE2D3 as a shared E2 — consistent with the redundancy already documented; not separately added (peripheral, not cached, RNF185-focused mechanism covered in the RNF185 review).
Thiadiazole RNF5 inhibitors (inh-2 / analogue 16; Brusa et al. 2023 J Med Chem, DOI 10.1021/acs.jmedchem.3c00608) and the 2024 CF ubiquitination-targeting review (Okiyoneda et al., DOI 10.1007/s00018-024-05295-z) are translational/medicinal-chemistry context only; no new functional annotation implied. Noted for completeness, not added as functional refs.
Falcon also restates the Ge & Zhang 2024 Front Immunol review (RNF5 negative regulator of STING/MAVS/IRF3; DOI 10.3389/fimmu.2023.1324516) — review-level, corroborates existing non-core antiviral annotations; not added.

Pn Notes

(RNF5-pn-notes.md)

RNF5 PN Consistency Notes

Generated: 2026-06-18
Project: PROTEOSTASIS
Scope: PN consistency rereview against local AIGR review and available deep-research artifacts
UniProt: Q99942
AIGR review status: COMPLETE
Review batch: proteostasis-batch-2026-06-11
Batch change status: added

Source Files Checked

AIGR review YAML: present - genes/human/RNF5/RNF5-ai-review.yaml
AIGR review HTML: missing - genes/human/RNF5/RNF5-ai-review.html
Gene notes: present - genes/human/RNF5/RNF5-notes.md
GOA TSV: present - genes/human/RNF5/RNF5-goa.tsv
UniProt record: present - genes/human/RNF5/RNF5-uniprot.txt
PN dossier: projects/PROTEOSTASIS/reports/phase1_dossiers/RNF5.md
PN consistency section: projects/PROTEOSTASIS/reports/phase1_dossiers/_sections/RNF5.md
PN projection report: projects/PROTEOSTASIS/reports/pn_projection/pn_projected_gene_go_summary.tsv
PN mapping audit: projects/PROTEOSTASIS/reports/pn_mapping_audit/current_mapping_scrutiny.tsv

Deep Research Files

genes/human/RNF5/RNF5-deep-research-falcon.md

AIGR Review Snapshot

Description: RNF5 (RMA1, NG2/G16) is a small (180 aa) tail-anchored RING-type E3 ubiquitin-protein ligase (EC 2.3.2.27) embedded in the endoplasmic reticulum membrane via two C-terminal transmembrane helices, with an N-terminal cytosolic C3HC4 RING domain (catalytic Cys-42) that recruits ubiquitin-charged E2 enzymes (notably the UBE2D/UbcH5 family and UBE2N/Ubc13). RNF5 is one of the founding mammalian ER-anchored ERAD ubiquitin ligases. Together with the E2 UBE2J1/Ubc6e and Derlin-1 it recognizes folding defects in membrane proteins co-translationally and assembles K48-linked polyubiquitin chains that commit misfolded clients such as CFTR and the disease-associated CFTR-deltaF508 mutant to retrotranslocation and proteasomal degradation; it functions partly redundantly with its close paralog RNF185, with which it forms an E3 ligase module central to CFTR degradation. Beyond canonical ERAD, RNF5 has documented ligase-dependent regulatory roles that use distinct chain topologies and substrates. It builds non-degradative K63-linked chains on the ERAD adaptor JKAMP/JAMP to limit its recruitment of proteasome and p97/VCP components; it ubiquitinates and degrades the innate-immune adaptor STING1/MITA at mitochondria to dampen antiviral type I interferon responses; it controls basal autophagy by regulating the stability of a membrane pool of the cysteine protease ATG4B; and it ubiquitinates paxillin to influence cell motility. RNF5 is widely expressed, and although localized predominantly to membranes (with reported plasma-membrane and mitochondrial-membrane pools), its ER-membrane localization underlies its core ERAD ligase function.
Existing/core annotation action counts: ACCEPT: 31; KEEP_AS_NON_CORE: 28; REMOVE: 3

PN Consistency Summary

Consistency: Excellent. Deep research, review and PN concur on the core: ER-membrane RING E3 (Cys-42), ERAD of CFTR/ΔF508 (K48), partly redundant with RNF185; plus the non-core ATG4B/autophagy, STING1/MITA antiviral (K48, mitochondria), JAMP (K63), and paxillin/motility roles. The PN ATG4 row's note ("controls stability of ATG4B") and its reference (PMID:23093945, ATG4B/RNF5 PLOS paper) match the review's GO:0061630 EXP annotation on PMID:23093945. No contradictions.
PN story / NEW pressure: GO:0061630 + GO:0036503 already in GOA and ACCEPTed (multiple EXP/IMP/IGI). GO:2000785 regulation of autophagosome assembly (verified real, OLS) is new_to_goa — RNF5 genuinely limits basal autophagy via ATG4B stability, so this is a DEFENSIBLE ADD, though it is a regulatory framing of a role the review treats as non-core moonlighting (the review captures ATG4B only as substrate-directed ligase activity, no autophagy-process term). Conclude: ligase MF + ERAD already captured; GO:2000785 defensible NEW (non-core).
Evidence alignment: Strong overlap. PN ATG4 row PMID:23093945 = review's autophagy annotation. PN RING row cites "19489725 / rev" (Kaneko/RING-family review) — NOT in the review's reference list (review uses gene-specific EXP papers instead); low impact since ligase MF is independently well-supported.
Verdict: Fully consistent; PN ligase/ERAD already captured, GO:2000785 a defensible (non-core) NEW autophagy-regulation term. No YAML change required.

Full Consistency Review

UniProt: Q99942 · batch: proteostasis-batch-2026-06-11 · review status: COMPLETE (very thorough; tail-anchored ER RING E3, ERAD core + STING/ATG4B/paxillin moonlighting)
PN placement: 3 rows — ER proteostasis|...|ER associated degradation|Cytosolic handling of ERAD substrates|ERAD-associated RING E3 ligase; ALP|Autophagophore initiation and elongation|...|Modulation of ATG4 activity; UPS|E3 ubiquitin and UBL ligases|RING|with transmembrane domain|ER, mitochondria, cell membrane. PN-node mapping: ERAD-RING-E3 subtype→GO:0061630 (in_goa); ERAD type/group→GO:0036503 (in_goa); ATG4-modulation subtype→GO:2000785 regulation of autophagosome assembly (new_to_goa); RING group→GO:0061630 (in_goa). Projected: GO:0036503×2, GO:0061630×2 (all in GOA), GO:2000785 (new).
Consistency: Excellent. Deep research, review and PN concur on the core: ER-membrane RING E3 (Cys-42), ERAD of CFTR/ΔF508 (K48), partly redundant with RNF185; plus the non-core ATG4B/autophagy, STING1/MITA antiviral (K48, mitochondria), JAMP (K63), and paxillin/motility roles. The PN ATG4 row's note ("controls stability of ATG4B") and its reference (PMID:23093945, ATG4B/RNF5 PLOS paper) match the review's GO:0061630 EXP annotation on PMID:23093945. No contradictions.
PN story / NEW pressure: GO:0061630 + GO:0036503 already in GOA and ACCEPTed (multiple EXP/IMP/IGI). GO:2000785 regulation of autophagosome assembly (verified real, OLS) is new_to_goa — RNF5 genuinely limits basal autophagy via ATG4B stability, so this is a DEFENSIBLE ADD, though it is a regulatory framing of a role the review treats as non-core moonlighting (the review captures ATG4B only as substrate-directed ligase activity, no autophagy-process term). Conclude: ligase MF + ERAD already captured; GO:2000785 defensible NEW (non-core).
Mapping strategy: Correct. Catalytic RING E3 → GO:0061630 at subtype/group; ERAD pathway at type/group; subtype-with-TM no_mapping avoids double-counting. ATG4-modulation leaf → GO:2000785 is appropriately conservative (regulation, not core autophagy). No node-status change warranted.
Evidence alignment: Strong overlap. PN ATG4 row PMID:23093945 = review's autophagy annotation. PN RING row cites "19489725 / rev" (Kaneko/RING-family review) — NOT in the review's reference list (review uses gene-specific EXP papers instead); low impact since ligase MF is independently well-supported.
Verdict: Fully consistent; PN ligase/ERAD already captured, GO:2000785 a defensible (non-core) NEW autophagy-regulation term. No YAML change required.
Recommended edits: none required. [YAML] (optional) consider adding GO:2000785 regulation of autophagosome assembly (IMP/IDA, PMID:23093945) as a non-core BP to mirror the PN projection. [REF] PN-cited PMID:19489725 (RING row) absent from review — verify it is a family review, not gene-specific.

PN Dossier Context

review_batch: proteostasis-batch-2026-06-11
review_yaml: genes/human/RNF5/RNF5-ai-review.yaml
PN workbook rows: 3

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

UniProt: Q99942
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 | Autophagophore initiation and elongation | ATG8 homolog processing, upstream | Preparation of ATG8 homologs for lipidation | Modulation of ATG4 activity

UniProt: Q99942
In branches: ER, ALP, UPS
Notes: Annotated by homology with mouse. Regulates basal levels of autophagy by controlling the stability of ATG4B
PN references (titles):
- Regulation of ATG4B Stability by RNF5 Limits Basal Levels of Autophagy and Influences Susceptibility to Bacterial Infection (plos.org)
PN-node mapping records (path + ancestors):
- [subtype] Autophagy-Lysosome Pathway|Autophagophore initiation and elongation|ATG8 homolog processing, upstream|Preparation of ATG8 homologs for lipidation|Modulation of ATG4 activity
  status=mapped scope=ok_for_propagation_to_go GO=[GO:2000785 regulation of autophagosome assembly]
  rationale: This PN leaf denotes factors that alter ATG4 function upstream of ATG8 processing. The cleanest GO-level consequence captured in the current ontology is regulation of autophagosome assembly.
- [type] Autophagy-Lysosome Pathway|Autophagophore initiation and elongation|ATG8 homolog processing, upstream|Preparation of ATG8 homologs for lipidation
  status=no_mapping scope= GO=[]
  rationale: Reviewed as a contextual PN role. The label is useful for curator triage, but by itself does not support a universal GO assertion for all member genes beyond curated ancestor or child mappings.
- [group] Autophagy-Lysosome Pathway|Autophagophore initiation and elongation|ATG8 homolog processing, upstream
  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|Autophagophore initiation and elongation
  status=context_only scope=too_broad_to_propagate GO=[GO:0016236 macroautophagy]
  rationale: This class is a real macroautophagy context, but its descendants include core factors, component buckets, upstream modulators, localization roles, and residual categories. Projecting generic macroautophagy from this ancestor creates TRAPP-like overpropagation, so candidate GO annotations must come from narrower curated nodes.
- [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, cell membrane

UniProt: Q99942
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, cell membrane
  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.

Projected GO annotations (5)

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:2000785 regulation of autophagosome assembly | scope=ok_for_propagation_to_go | goa_status=new_to_goa | from=Autophagy-Lysosome Pathway|Autophagophore initiation and elongation|ATG8 homolog processing, upstream|Preparation of ATG8 homologs for lipidation|Modulation of ATG4 activity
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

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: Q99942
gene_symbol: RNF5
product_type: PROTEIN
status: COMPLETE
taxon:
  id: NCBITaxon:9606
  label: Homo sapiens
description: >-
  RNF5 (RMA1, NG2/G16) is a small (180 aa) tail-anchored RING-type E3
  ubiquitin-protein ligase (EC 2.3.2.27) embedded in the endoplasmic reticulum
  membrane via two C-terminal transmembrane helices, with an N-terminal
  cytosolic C3HC4 RING domain (catalytic Cys-42) that recruits ubiquitin-charged
  E2 enzymes (notably the UBE2D/UbcH5 family and UBE2N/Ubc13). RNF5 is one of
  the founding mammalian ER-anchored ERAD ubiquitin ligases. Together with the
  E2 UBE2J1/Ubc6e and Derlin-1 it recognizes folding defects in membrane
  proteins co-translationally and assembles K48-linked polyubiquitin chains that
  commit misfolded clients such as CFTR and the disease-associated CFTR-deltaF508
  mutant to retrotranslocation and proteasomal degradation; it functions partly
  redundantly with its close paralog RNF185, with which it forms an E3 ligase
  module central to CFTR degradation. Beyond canonical ERAD, RNF5 has documented
  ligase-dependent regulatory roles that use distinct chain topologies and
  substrates. It builds non-degradative K63-linked chains on the ERAD adaptor
  JKAMP/JAMP to limit its recruitment of proteasome and p97/VCP components; it
  ubiquitinates and degrades the innate-immune adaptor STING1/MITA at
  mitochondria to dampen antiviral type I interferon responses; it controls
  basal autophagy by regulating the stability of a membrane pool of the cysteine
  protease ATG4B; and it ubiquitinates paxillin to influence cell motility. RNF5
  is widely expressed, and although localized predominantly to membranes (with
  reported plasma-membrane and mitochondrial-membrane pools), its ER-membrane
  localization underlies its core ERAD ligase function.
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 RNF5, conserved across the RNF5/RNF185 family.
    action: ACCEPT
    reason: Core molecular function; supported experimentally (EC 2.3.2.27; C42S abolishes activity) and across the family.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
- 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 RNF5 as an ER-anchored ERAD ligase.
    action: ACCEPT
    reason: Core biological process; directly supported by IMP evidence (CFTR/CFTR-deltaF508 degradation) and conserved in the family.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: Cystic fibrosis transmembrane conductance regulator (CFTR) is one ERAD substrate targeted to co-translational degradation by the E3 ligase RNF5/RMA1
- term:
    id: GO:0044390
    label: ubiquitin-like protein conjugating enzyme binding
  evidence_type: IBA
  original_reference_id: GO_REF:0000033
  qualifier: enables
  review:
    summary: RNF5 binds ubiquitin-conjugating (E2) enzymes, including the UBE2D/UbcH5 family and UBE2N/Ubc13, as part of its catalytic cycle. This is informative but ancillary to the ligase activity itself.
    action: KEEP_AS_NON_CORE
    reason: Accurately reflects E2 binding (UBE2D1/UBE2D2, UBE2N) required for catalysis, but is a mechanistic subsidiary of the core ubiquitin ligase activity.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: May function together with E2 ubiquitin-conjugating enzymes UBE2D1/UBCH5A and UBE2D2/UBC4
- 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 RNF5's ER-membrane site of ERAD function.
    action: ACCEPT
    reason: Correct compartment; redundant with the more specific ER membrane annotations and IDA (HPA) evidence.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Endoplasmic reticulum membrane {ECO:0000269|PubMed:19285439}'
- 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; this is the core compartment for RNF5's ERAD ligase function.
    action: ACCEPT
    reason: Correct core localization; RNF5 is a tail-anchored ER membrane protein, supported experimentally (EXP, PMID:19285439).
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Endoplasmic reticulum membrane {ECO:0000269|PubMed:19285439}'
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: Electronic transfer of plasma membrane localization from the original cloning study (PMID:9533025). A real but secondary localization not tied to the core ERAD function.
    action: KEEP_AS_NON_CORE
    reason: Reported in the early cloning paper but the functionally dominant compartment for RNF5 is the ER membrane; plasma membrane pool is peripheral.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Cell membrane {ECO:0000269|PubMed:9533025}'
- 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 RNF5'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:24019521
      supporting_text: degraded by the ubiquitin-proteasome pathway through a process called ER-associated degradation (ERAD)
- term:
    id: GO:0031966
    label: mitochondrial membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000044
  qualifier: located_in
  review:
    summary: Electronic transfer of mitochondrial membrane localization, reflecting the antiviral STING1/MITA study in which RNF5 acts at mitochondria. A real but secondary, immunity-related localization.
    action: KEEP_AS_NON_CORE
    reason: Supported by PMID:19285439 (MITA ubiquitination at mitochondria) but represents a secondary, context-specific role distinct from the core ER-membrane ERAD function.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: virus-induced ubiquitination and degradation of MITA by RNF5 occurred at the mitochondria
- 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 and IBA evidence.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: Cystic fibrosis transmembrane conductance regulator (CFTR) is one ERAD substrate targeted to co-translational degradation by the E3 ligase RNF5/RMA1
- 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 RNF5 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:24019521
      supporting_text: hosts a machinery called ERQC for ER quality control
- 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 EXP/IBA evidence.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-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. RNF5 is a ubiquitin ligase and does not trim mannose; this is a pathway-adjacency over-annotation propagated from the ERAD/ERQC context.
    action: REMOVE
    reason: RNF5 has no glycosidase/mannosidase activity; mannose trimming is performed by EDEM/ER mannosidases, not by the E3 ligase. This electronic inference is biologically incorrect.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:12861019
  qualifier: enables
  review:
    summary: IntAct interaction with paxillin (PXN), a functionally meaningful RNF5 substrate, but the bare protein binding term is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Records a real substrate interaction (PXN) but bare protein binding is uninformative per curation guidelines.
    supported_by:
    - reference_id: PMID:12861019
      supporting_text: the human homologue of RNF5 associates with the amino-terminal domain of paxillin
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:14667819
  qualifier: enables
  review:
    summary: High-throughput yeast two-hybrid interactions (e.g. ABHD16A, UBE2 enzymes). Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Records real IntAct interactions but bare protein binding is uninformative and not a core function.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; O95870: ABHD16A'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:16901789
  qualifier: enables
  review:
    summary: Interaction with CFTR captured in the sequential-triage ERAD study; CFTR is a key RNF5 ERAD substrate. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Records the functionally important RNF5-CFTR interaction, but bare protein binding is uninformative; the substrate relationship is captured by the ERAD process annotations.
    supported_by:
    - reference_id: PMID:16901789
      supporting_text: an ER membrane-associated ubiquitin ligase complex containing the E3 RMA1, the E2 Ubc6e, and Derlin-1
- 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 RNF5 binding to multiple UBE2D/UBE2E/UBE2W E2s. 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/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; P51668: UBE2D1'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:20152160
  qualifier: enables
  review:
    summary: Interaction with the E2 UBE2D2/UbcH5b from a structural study of the E2~Ub conjugate. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Real E2 interaction but bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; P62837: UBE2D2'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25416956
  qualifier: enables
  review:
    summary: Large-scale interactome map capturing numerous RNF5 partners (including SLC transporters, RNF185, SEC22A). Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: High-throughput interactome; bare protein binding is uninformative and not a core function.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q96GF1: RNF185'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:25759021
  qualifier: enables
  review:
    summary: Interactions with glutamine carrier proteins (SLC1A5, SLC38A2) relevant to RNF5's role in breast cancer ER-stress response. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Records real substrate/carrier interactions but bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q15758: SLC1A5'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:26618866
  qualifier: enables
  review:
    summary: Interaction with CFTR from a deltaF508 CFTR interactome-remodeling study. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Real CFTR interaction relevant to ERAD but bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; P13569: CFTR'
- 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/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; 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 RNF5 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/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q9Y5U4: INSIG2'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:32814053
  qualifier: enables
  review:
    summary: Interactions (e.g. OPTN, UBE2K) from a neurodegenerative-disease interactome study. 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/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q96CV9: OPTN'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:33961781
  qualifier: enables
  review:
    summary: Cell-specific interactome capturing RNF5 partners including RNF185 and RHBDD1. 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/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q96GF1: RNF185'
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:36012204
  qualifier: enables
  review:
    summary: Proximity-labeling interaction with CFTR identifying enrichment in SLC transporters. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Real CFTR/SLC interactions but bare protein binding is uninformative.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; P13569: CFTR'
- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:25416956
  qualifier: enables
  review:
    summary: RNF5 self-interaction captured in a proteome-scale interactome. Reflects homo-association seen in screens.
    action: KEEP_AS_NON_CORE
    reason: Documents RNF5 self-association (IntAct EBI-348482 with itself) but is peripheral to the core ligase function.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q99942: RNF5'
- term:
    id: GO:0042802
    label: identical protein binding
  evidence_type: IPI
  original_reference_id: PMID:32296183
  qualifier: enables
  review:
    summary: RNF5 self-interaction captured in the binary interactome reference map.
    action: KEEP_AS_NON_CORE
    reason: Documents RNF5 self-association but is peripheral to the core ligase function.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Q99942; Q99942: RNF5'
- term:
    id: GO:0016020
    label: membrane
  evidence_type: IEA
  original_reference_id: GO_REF:0000107
  qualifier: located_in
  review:
    summary: Ortholog-based electronic assignment of generic membrane localization, a parent of the specific ER membrane localization.
    action: KEEP_AS_NON_CORE
    reason: Correct but generic; subsumed by the more specific ER membrane localization.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: Multi-pass membrane protein
- 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). RNF5 is a ubiquitin ligase, not a transporter; this reflects pathway context (CFTR/ABC transport) bleed-through.
    action: REMOVE
    reason: RNF5 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, not a direct function.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
- 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-associated ubiquitination RNF5 performs.
    action: KEEP_AS_NON_CORE
    reason: Correct but generic; the specific ERAD pathway and K48/K63 ubiquitination annotations better capture the role.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-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 RNF5 (with RNF185) 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/RNF5/RNF5-uniprot.txt
      supporting_text: Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
- term:
    id: GO:0005783
    label: endoplasmic reticulum
  evidence_type: IDA
  original_reference_id: GO_REF:0000052
  qualifier: located_in
  review:
    summary: Direct immunofluorescence (HPA) evidence for ER localization, consistent with RNF5'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: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Endoplasmic reticulum membrane {ECO:0000269|PubMed:19285439}'
- term:
    id: GO:0005789
    label: endoplasmic reticulum membrane
  evidence_type: EXP
  original_reference_id: PMID:19285439
  qualifier: located_in
  review:
    summary: Experimental evidence that RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (ER)
- term:
    id: GO:0005886
    label: plasma membrane
  evidence_type: EXP
  original_reference_id: PMID:9533025
  qualifier: located_in
  review:
    summary: Experimental localization to the plasma membrane reported in the original cloning study. A real but secondary localization.
    action: KEEP_AS_NON_CORE
    reason: Documented in the cloning paper but the functionally dominant compartment is the ER membrane; plasma-membrane pool is peripheral to the core ERAD role.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: 'Predominantly located in the plasma membrane, with some localization occurring within cytoplasmic organelles'
- term:
    id: GO:0031966
    label: mitochondrial membrane
  evidence_type: EXP
  original_reference_id: PMID:19285439
  qualifier: located_in
  review:
    summary: Experimental localization to the mitochondrial membrane, where RNF5 ubiquitinates STING1/MITA during antiviral responses. A real but secondary, immunity-related localization.
    action: KEEP_AS_NON_CORE
    reason: Directly supported (PMID:19285439) but a secondary compartment for a context-specific immune role, not the core ER ERAD function.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: virus-induced ubiquitination and degradation of MITA by RNF5 occurred at the mitochondria
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: EXP
  original_reference_id: PMID:19269966
  qualifier: enables
  review:
    summary: Experimental demonstration of RNF5 ubiquitin ligase activity (RING-dependent ubiquitination of JAMP). Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental support; WT but not RING-mutant RNF5 ubiquitinates substrate.
    supported_by:
    - reference_id: PMID:19269966
      supporting_text: Ectopically expressed JAMP was efficiently ubiquitinated in vivo by WT but not the RING mutant form of RNF5
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: EXP
  original_reference_id: PMID:19285439
  qualifier: enables
  review:
    summary: Experimental demonstration of RNF5 ligase activity ubiquitinating STING1/MITA. Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental support.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: RNF5 targeted MITA at Lys150 for ubiquitination and degradation after viral infection
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: EXP
  original_reference_id: PMID:23093945
  qualifier: enables
  review:
    summary: Experimental demonstration of RNF5 ligase activity controlling ATG4B ubiquitination and stability. Core molecular function.
    action: ACCEPT
    reason: Core molecular function with direct experimental support.
    supported_by:
    - reference_id: PMID:23093945
      supporting_text: the membrane-associated E3 ligase RNF5 regulates basal levels of autophagy by controlling the stability of a select pool of the cysteine protease ATG4B
- term:
    id: GO:0006511
    label: ubiquitin-dependent protein catabolic process
  evidence_type: IGI
  original_reference_id: PMID:24019521
  qualifier: involved_in
  review:
    summary: Genetic-interaction evidence (with RNF185) that RNF5 drives ubiquitin-dependent degradation of CFTR; redundant depletion blocks CFTR-deltaF508 turnover.
    action: ACCEPT
    reason: Supported by the RNF5/RNF185 co-depletion experiments; consistent with the core ERAD/proteasomal degradation role.
    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:0036503
    label: ERAD pathway
  evidence_type: IMP
  original_reference_id: PMID:24019521
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence that RNF5 is required for ERAD of CFTR/CFTR-deltaF508. Core biological process.
    action: ACCEPT
    reason: Core biological process with direct experimental (IMP) support from depletion experiments.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: simultaneous depletion of RNF5 and RNF185 profoundly blocks CFTRΔF508 degradation
- term:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  evidence_type: TAS
  original_reference_id: PMID:24019521
  qualifier: enables
  review:
    summary: Author statement of RNF5 ubiquitin ligase activity in CFTR ERAD. Core molecular function.
    action: ACCEPT
    reason: Correct core molecular function.
    supported_by:
    - reference_id: PMID:24019521
      supporting_text: targeted to co-translational degradation by the E3 ligase RNF5/RMA1
- 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. RNF5 does not perform mannose trimming; this is pathway-adjacency over-annotation.
    action: REMOVE
    reason: RNF5 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/RNF5/RNF5-uniprot.txt
      supporting_text: Membrane-bound E3 ubiquitin-protein ligase that mediates ubiquitination of target proteins
- term:
    id: GO:0044877
    label: protein-containing complex binding
  evidence_type: IPI
  original_reference_id: PMID:24019521
  qualifier: enables
  review:
    summary: RNF5 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:0005789
    label: endoplasmic reticulum membrane
  evidence_type: TAS
  original_reference_id: Reactome:R-HSA-8866542
  qualifier: located_in
  review:
    summary: Reactome curation of RNF5 ER membrane localization within CFTR ERAD reactions. Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 ER membrane localization (CFTR binds ERAD machinery). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 ER membrane localization (CFTR F508del translocation). Core compartment.
    action: ACCEPT
    reason: Correct core localization; redundant with experimental evidence.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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 RNF5 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:19285439
      supporting_text: Both MITA and RNF5 were located at the mitochondria and endoplasmic reticulum (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:0030163
    label: protein catabolic process
  evidence_type: IMP
  original_reference_id: PMID:19285439
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence that RNF5 drives degradation of a target protein (STING1/MITA). A parent of the more specific ubiquitin-dependent catabolic/ERAD process.
    action: KEEP_AS_NON_CORE
    reason: Correct but generic relative to the specific ERAD and K48-ubiquitination annotations; here the catabolic target is the immune adaptor MITA.
    supported_by:
    - reference_id: PMID:19285439
      supporting_text: RNF5 targeted MITA at Lys150 for ubiquitination and degradation after viral infection
- term:
    id: GO:0070936
    label: protein K48-linked ubiquitination
  evidence_type: IDA
  original_reference_id: PMID:19285439
  qualifier: involved_in
  review:
    summary: Direct evidence that RNF5 builds K48-linked polyubiquitin chains (degradative topology) on STING1/MITA. This degradative topology is also used on CFTR.
    action: ACCEPT
    reason: Directly demonstrated K48-linked ubiquitination, the canonical degradative topology underlying RNF5's ERAD and MITA-degradation roles.
    supported_by:
    - reference_id: PMID:19269966
      supporting_text: CFTR ubiquitination occurred at the canonical Lys-48 topology
- term:
    id: GO:0004842
    label: ubiquitin-protein transferase activity
  evidence_type: IDA
  original_reference_id: PMID:19269966
  qualifier: enables
  review:
    summary: Direct evidence 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 RING E3 ligase activity.
    supported_by:
    - reference_id: PMID:19269966
      supporting_text: Ectopically expressed JAMP was efficiently ubiquitinated in vivo by WT but not the RING mutant form of RNF5
- term:
    id: GO:0005515
    label: protein binding
  evidence_type: IPI
  original_reference_id: PMID:19269966
  qualifier: enables
  review:
    summary: Interaction with JKAMP/JAMP, a functionally important RNF5 substrate/partner at the ER membrane. Bare protein binding is uninformative.
    action: KEEP_AS_NON_CORE
    reason: Records the real RNF5-JAMP interaction but bare protein binding is uninformative.
    supported_by:
    - reference_id: PMID:19269966
      supporting_text: RNF5 associates with JAMP in the ER membrane
- term:
    id: GO:0036503
    label: ERAD pathway
  evidence_type: IMP
  original_reference_id: PMID:19269966
  qualifier: involved_in
  review:
    summary: Mutant-phenotype evidence linking RNF5 to ERAD, here via regulation of the ERAD adaptor JAMP. Core biological process.
    action: ACCEPT
    reason: Core biological process; RNF5 modulates ERAD both by direct substrate ubiquitination and by K63-regulation of JAMP.
    supported_by:
    - reference_id: PMID:19269966
      supporting_text: RNF5 is a ubiquitin ligase anchored to the ER membrane implicated in ERAD via ubiquitination of misfolded proteins
- term:
    id: GO:0070534
    label: protein K63-linked ubiquitination
  evidence_type: IDA
  original_reference_id: PMID:19269966
  qualifier: involved_in
  review:
    summary: Direct evidence that RNF5 builds non-degradative K63-linked polyubiquitin chains on JAMP (Ubc13/UBE2N-dependent), a regulatory rather than degradative modification.
    action: ACCEPT
    reason: Directly demonstrated K63-linked ubiquitination, establishing RNF5's capacity for non-canonical regulatory ubiquitination distinct from K48 degradative chains.
    supported_by:
    - reference_id: PMID:19269966
      supporting_text: These findings establish that RNF5 mediates Lys-63-based polyubiquitination of JAMP
- term:
    id: GO:0008270
    label: zinc ion binding
  evidence_type: TAS
  original_reference_id: PMID:9533025
  qualifier: enables
  review:
    summary: The C3HC4 RING domain coordinates zinc as a structural requirement for the ligase fold. A structural attribute supporting, not equal to, the ligase activity.
    action: KEEP_AS_NON_CORE
    reason: Accurate structural feature of the RING domain (residues 27-68) but subsidiary to the informative ubiquitin ligase activity; not a standalone core function.
    supported_by:
    - reference_id: file:human/RNF5/RNF5-uniprot.txt
      supporting_text: RING-type
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:0000052
  title: Gene Ontology annotation based on curation of immunofluorescence data
  findings: []
- id: GO_REF:0000107
  title: Automatic transfer of experimentally verified manual GO annotation data to orthologs using Ensembl Compara
  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:12861019
  title: RNF5, a RING finger protein that regulates cell motility by targeting paxillin ubiquitination and altered localization.
  findings:
  - statement: RNF5 associates with the N-terminal domain of paxillin and mediates its ubiquitination, requiring intact RING and C-terminal domains, regulating cell motility.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Establishes a non-core cell-motility role via paxillin ubiquitination; the source of the GO:0005515 PXN interaction.
- id: PMID:14667819
  title: Analysis of a high-throughput yeast two-hybrid system and its use to predict the function of intracellular proteins encoded within the human MHC class III region.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: High-throughput Y2H interactome (MHC class III region); source of several bare protein binding annotations.
- id: PMID:16901789
  title: Sequential quality-control checkpoints triage misfolded cystic fibrosis transmembrane conductance regulator.
  findings:
  - statement: An ER membrane-associated ubiquitin ligase complex of E3 RMA1/RNF5, E2 Ubc6e and Derlin-1 recognizes folding defects in CFTR/CFTR-deltaF508 co-translationally and cooperates with cytosolic Hsc70/CHIP to promote proteasomal degradation.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Foundational study establishing RNF5/RMA1 as a co-translational ER-membrane ERAD ligase for CFTR.
- id: PMID:19269966
  title: Regulation of endoplasmic reticulum-associated degradation by RNF5-dependent ubiquitination of JNK-associated membrane protein (JAMP).
  findings:
  - statement: RNF5 builds Ubc13(UBE2N)-dependent K63-linked polyubiquitin chains on the ERAD adaptor JAMP at the ER membrane, reducing JAMP association with proteasome subunits and p97/VCP and thereby limiting ERAD; RNF5 builds canonical K48 chains on CFTR.
    reference_section_type: RESULTS
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Full text available; demonstrates RNF5 RING-dependent ligase activity and distinct K48 (CFTR) vs K63 (JAMP) chain topologies.
- id: PMID:19285439
  title: The ubiquitin ligase RNF5 regulates antiviral responses by mediating degradation of the adaptor protein MITA.
  findings:
  - statement: RNF5 interacts with STING1/MITA in a virus-infection-dependent manner and builds K48-linked chains on MITA at Lys150 for proteasomal degradation at mitochondria, negatively regulating type I interferon antiviral responses; RNF5 localizes at ER and mitochondria.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Establishes the non-core antiviral/immunity role and mitochondrial localization; source of K48 ubiquitination and mitochondrial-membrane annotations.
- 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 RNF5 binding to multiple UBE2D/UBE2E/UBE2W enzymes relevant to its catalytic cycle.
- id: PMID:20152160
  title: 'Crystal structure of UbcH5b~ubiquitin intermediate: insight into the formation of the self-assembled E2~Ub conjugates.'
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Structural E2~Ub study; captures RNF5-UBE2D2 interaction.
- id: PMID:23093945
  title: Regulation of ATG4B stability by RNF5 limits basal levels of autophagy and influences susceptibility to bacterial infection.
  findings:
  - statement: RNF5 regulates basal autophagy by controlling the stability of a membrane pool of the cysteine protease ATG4B, thereby limiting LC3/ATG8 processing required for autophagosome formation.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Establishes the non-core autophagy-regulatory role via ATG4B; provides EXP support for ligase activity.
- 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 and RNF5 form a partly redundant E3 ligase module central to CFTR/CFTR-deltaF508 degradation; co-depletion profoundly blocks CFTR-deltaF508 degradation during and after translation.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: HIGH
    correctness: VERIFIED
    review_notes: Full text available; establishes RNF5/RNF185 redundancy in CFTR ERAD; source of IMP/IGI ERAD 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 and identical protein binding (self) annotations.
- id: PMID:25759021
  title: Regulation of glutamine carrier proteins by RNF5 determines breast cancer response to ER stress-inducing chemotherapies.
  findings:
  - statement: RNF5 regulates glutamine carrier proteins (SLC1A5, SLC38A2) under ER stress, influencing breast cancer response to chemotherapy.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Non-core cancer/ER-stress role; source of SLC transporter interactions.
- id: PMID:26618866
  title: ∆F508 CFTR interactome remodelling promotes rescue of cystic fibrosis.
  findings: []
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: CFTR-deltaF508 interactome study; captures RNF5-CFTR interaction relevant to ERAD.
- 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 and self-interaction annotations.
- id: PMID:32814053
  title: Interactome Mapping Provides a Network of Neurodegenerative Disease Proteins and Uncovers Widespread Protein Aggregation in Affected Brains.
  findings: []
  reference_review:
    relevance: LOW
    correctness: VERIFIED
    review_notes: Neurodegeneration interactome; source of bare protein binding annotations (e.g. OPTN, UBE2K).
- 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 (RNF185, RHBDD1).
- id: PMID:36012204
  title: Differential CFTR-Interactome Proximity Labeling Procedures Identify Enrichment in Multiple SLC Transporters.
  findings: []
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: CFTR proximity-labeling interactome; captures RNF5-CFTR interaction relevant to ERAD.
- id: PMID:9533025
  title: Cloning, expression and mapping of a novel RING-finger gene (RNF5), a human homologue of a putative zinc-finger gene from Caenorhabditis elegans.
  findings:
  - statement: Original cloning of RNF5, a RING-finger gene in the MHC class III region; reported widespread expression and predominantly plasma-membrane localization with some cytoplasmic-organelle localization.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: Original cloning/characterization; source of zinc ion binding (TAS) and plasma-membrane localization annotations.
- 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:39098896
  title: RING finger protein 5 protects against acute myocardial infarction by inhibiting ASK1.
  findings:
  - statement: RNF5 is downregulated in infarcted heart tissue; RNF5 knockout worsens and RNF5 overexpression ameliorates myocardial infarction injury in mouse and cardiomyocyte models, with protection attributed to inhibition of ASK1 (MAP3K5) activation.
    reference_section_type: ABSTRACT
  reference_review:
    relevance: MEDIUM
    correctness: VERIFIED
    review_notes: PubMed-verified (PMID:39098896 = BMC Cardiovasc Disord 2024, DOI 10.1186/s12872-024-04070-z); proposes a non-core cardioprotective role for RNF5 via the ASK1 stress-signaling pathway. Directness of RNF5 E3 action on ASK1 not fully resolved; 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 ubiquitinates misfolded membrane-protein clients (e.g. CFTR and CFTR-deltaF508) with K48-linked chains to commit them to retrotranslocation and proteasomal degradation via the ERAD pathway, acting partly redundantly with RNF185.
  molecular_function:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  locations:
  - id: GO:0005789
    label: endoplasmic reticulum membrane
  supported_by:
  - reference_id: PMID:16901789
    supporting_text: an ER membrane-associated ubiquitin ligase complex containing the E3 RMA1, the E2 Ubc6e, and Derlin-1
  - 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-membrane ubiquitin ligase that builds non-degradative K63-linked polyubiquitin chains on the ERAD adaptor JKAMP/JAMP (UBE2N/Ubc13-dependent), modulating its recruitment of proteasome and p97/VCP components and thereby regulating ERAD flux.
  molecular_function:
    id: GO:0061630
    label: ubiquitin protein ligase activity
  locations:
  - id: GO:0005789
    label: endoplasmic reticulum membrane
  supported_by:
  - reference_id: PMID:19269966
    supporting_text: These findings establish that RNF5 mediates Lys-63-based polyubiquitination of JAMP
  directly_involved_in:
  - id: GO:0070534
    label: protein K63-linked ubiquitination
proposed_new_terms: []
suggested_questions:
- question: What determines RNF5 substrate selectivity and chain-topology choice (K48 degradative on CFTR/MITA vs K63 regulatory on JAMP), and which E2 partner (UBE2D vs UBE2N) is decisive in each case?
- question: To what extent are RNF5's antiviral (STING1/MITA) and autophagy (ATG4B) roles separable from its core ER ERAD function, and do they require distinct subcellular pools?
suggested_experiments:
- description: Reconstitute ERAD ubiquitination of CFTR-deltaF508 in vitro with purified RNF5, RNF185, UBE2J1/Ubc6e and Derlin-1 to map lysine sites and ubiquitin-chain linkage on the substrate and to dissect RNF5/RNF185 redundancy.
- description: Generate RNF5 and RNF5/RNF185 double-knockout cells and perform quantitative ubiquitinome/proteome profiling under basal and ER-stress conditions to define the endogenous ERAD substrate repertoire and the degree of paralog redundancy.

RNF5

Gene Description

Existing Annotations Review

Core Functions

RING-type E3 ubiquitin ligase anchored in the ER membrane that ubiquitinates misfolded membrane-protein clients (e.g. CFTR and CFTR-deltaF508) with K48-linked chains to commit them to retrotranslocation and proteasomal degradation via the ERAD pathway, acting partly redundantly with RNF185.

ER-membrane ubiquitin ligase that builds non-degradative K63-linked polyubiquitin chains on the ERAD adaptor JKAMP/JAMP (UBE2N/Ubc13-dependent), modulating its recruitment of proteasome and p97/VCP components and thereby regulating ERAD flux.

References

Suggested Questions for Experts

Suggested Experiments

Deep Research

Falcon

Research report: Human RNF5 (UniProt Q99942) functional annotation (2023–2024 emphasis)

0) Target verification (critical)

1) Key concepts and definitions (current understanding)

1.1 Ubiquitination and E3 ligases

1.2 ER-associated degradation (ERAD) and ER quality control (ERQC)

1.3 Innate immune adaptor regulation (cGAS–STING and RIG-I/MAVS)

2) RNF5 molecular function, localization, and substrate specificity

2.1 Subcellular localization

2.2 RNF5 in CFTR ERAD: substrate recognition and pathway placement

2.3 Innate immunity substrates: STING, MAVS, IRF3

2.4 Autophagy-related regulation (ATG4B)

2.5 Cardiovascular signaling (ASK1)

3) Recent developments (prioritizing 2023–2024)

3.1 Genome-wide CRISPR screens redefine RNF5’s role as important but partially redundant in CFTR-F508del ERAD (2024)

3.2 Medicinal chemistry progress: thiadiazole RNF5 inhibitors that enhance mutant CFTR rescue (2023)

3.3 Updated CF translational assessment: no ubiquitination-pathway drugs in CF clinical trials (2024 review)

3.4 RNF5 as an innate-immunity node and potential host-directed antiviral target (2024 review)

4) Current applications and real-world implementations

4.1 Cystic fibrosis: modulating RNF5-mediated ubiquitination to rescue CFTR

4.2 Host-directed antiviral strategies

4.3 Cardiovascular disease models

5) Expert opinions and analysis (authoritative perspectives)

5.1 CF field perspective: redundancy and risk-benefit constraints

5.2 Systems view from CRISPR genetics: RNF5 is a major node but not sufficient alone

5.3 Innate immunity perspective: RNF5 as a negative regulator of STING/MAVS

6) Relevant statistics and data points (recent studies)

7) Mechanistic summary figure

8) Summary and functional annotation statement

Appendix: compact evidence map

Artifacts

Citations

📚 Additional Documentation

Notes

RNF5 (RMA1) review notes

Core identity

Primary role: ERAD of misfolded membrane proteins (incl. CFTR/CFTRdeltaF508)

Functional redundancy with RNF185

E2 partners / ubiquitin chain topologies

Secondary / non-core roles (pleiotropic)

Localization

Annotation review decisions (summary)

Falcon deep-research findings (incorporated 2026-06)

Pn Notes

RNF5 PN Consistency Notes

Source Files Checked

Deep Research Files

AIGR Review Snapshot

PN Consistency Summary

Full Consistency Review

PN Dossier Context

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

PN row 2: Autophagy-Lysosome Pathway | Autophagophore initiation and elongation | ATG8 homolog processing, upstream | Preparation of ATG8 homologs for lipidation | Modulation of ATG4 activity

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

Projected GO annotations (5)

Note

📄 View Raw YAML