| Aspect | Key finding | Experimental evidence/assay | Key quantitative/statistical notes | Primary source (include first author year journal) and URL | Citation ID |
|---|---|---|---|---|---|
| Molecular function | RNF185 is a RING-dependent ERAD E3 ligase localized predominantly to the ER; it associates with ERAD factors and shows E2 preference for UbcH5c in vitro, with cell-based association to Ubc6e/UBE2J1 and UBE2J2 | Purified GST-RNF185 in vitro auto-ubiquitination with E1/E2/ubiquitin; co-immunoprecipitation with Derlin-1, Erlin2, Ubc6e/UBE2J1, UBE2J2; RING mutants and TM truncation to test catalytic and localization requirements | Robust auto-ubiquitination with UbcH5c, weaker with UbcH6, none with UbcH7; C39/C42 RING mutations or RING deletion abolished activity; distal TM truncation disrupted ER targeting; tunicamycin induced RNF185 transcripts peaking at ~12 h | Khouri 2013, *Journal of Biological Chemistry*. https://doi.org/10.1074/jbc.m113.470500 | (pqac-00000000) |
| Substrate / phenotype | RNF185 targets CFTR and CFTR-ΔF508 for ubiquitin-proteasome degradation, acting with RNF5 as a major ERAD module for co- and post-translational control | Pulse-labeling with [35S]Met/Cys, cycloheximide chase, proteasome inhibition (ALLN), overexpression and knockdown/silencing, anti-HA IP and quantification | RNF185 overexpression reduced CFTR-F508 half-life from 44 to 29 min and reduced 35S-labeled CFTR signal by up to ~50%; RNF185 knockdown increased labeled CFTR at least ~2-fold; RNF5 knockdown ~3-fold stabilization, RNF185 knockdown ~2-fold, combined depletion ~4.5-fold stabilization of CFTR-F508 steady-state levels | Khouri 2013, *Journal of Biological Chemistry*. https://doi.org/10.1074/jbc.m113.470500 | (pqac-00000004, pqac-00000007, pqac-00000020, pqac-00000021) |
| Localization / pathway | RNF185 is a mitochondrial outer membrane E3 ligase; its two C-terminal TM domains target it to mitochondria and it promotes selective mitochondrial autophagy/mitophagy via BNIP1 | Confocal colocalization with MitoTracker and DsRed2-Mito; biochemical fractionation; proteinase K protection; GFP-LC3 puncta assays; LC3-I to LC3-II immunoblotting; siRNA knockdown; in vivo ubiquitination assays | TM2 especially required for mitochondrial targeting; RNF185 overexpression increased LC3-II and GFP-LC3 puncta; RNF185 knockdown reduced basal LC3-II; BNIP1 was polyubiquitinated by RNF185 through K63-linked chains and recruited p62 | Tang 2011, *PLoS ONE*. https://doi.org/10.1371/journal.pone.0024367 | (pqac-00000005, pqac-00000006) |
| Substrate / pathway | In ebolavirus infection, ER-associated RNF185 polyubiquitinates EBOV GP1,2 on Lys673 with K27-linked chains, diverting it to SQSTM1/p62-dependent reticulophagy/ERLAD rather than proteasomal degradation | Infection-based proteostasis assays; ubiquitination mapping; dependence on SQSTM1/p62, ATG3, and ATG5; ER proteostasis framework linking calnexin cycle, ERAD, and ERLAD | K27-linked polyubiquitination on GP1,2 Lys673; degradation proceeds via lysosome/autophagosome recruitment rather than proteasome; study concludes this increases viral fitness | Zhang 2022, *Nature Communications*. https://doi.org/10.1038/s41467-022-33805-9 | (pqac-00000001) |
| Substrate / pathway / phenotype | RNF185 forms an ERAD complex with Membralin (MBRL) that recognizes unassembled tapasin (TPSN), promotes its ubiquitination and degradation, and thereby limits MHC-I surface expression | Quantitative proteomics (TMT-LC-MS/MS); IP-MS and co-IP; cycloheximide chase; ubiquitination assays; knockout/rescue in iPSCs, HEK293, THP-1, U2OS, and iPSC-derived macrophages; W6/32 flow cytometry for MHC-I | TPSN half-life ~4 h in control cells; RNF185 or MBRL loss increased TPSN steady-state levels and reduced ubiquitinated TPSN despite higher total TPSN; TPSN tail 4K→A mutant poorly ubiquitinated; p<0.05 threshold in proteomics; n=5 parental and n=5 MBRL KO astrocyte samples in TMT experiment; IFNγ stimulation used at 100 ng/mL for 16 h; CHX quantification from n=3 experiments | van de Weijer 2024, *Nature Communications*. https://doi.org/10.1038/s41467-024-52772-x | (pqac-00000008, pqac-00000009, pqac-00000010, pqac-00000011, pqac-00000012, pqac-00000014, pqac-00000015) |
| Substrate / pathway | During Senecavirus A infection, RNF185 catalyzes K27-linked polyubiquitination of mitochondrial TUFM, enabling SQSTM1 recognition and mitophagy that promotes viral replication | Co-IP and ubiquitination assays with HA-Ub and Ub lysine mutants; deletion-mutant mapping; docking/residue mapping; siRNA knockdown and overexpression; western blot/autophagy-marker analysis; TCID50 viral titration | Only K27R ubiquitin mutant significantly reduced TUFM polyubiquitination; RNF185 interacts with TUFM via RNF185 TM1 (aa133–155); TUFM interaction regions mapped mainly to aa56–252 and around aa344/345; RNF185 knockdown increased TUFM, SQSTM1, TIMM23, TOMM20 and decreased LC3-II; experiments performed in three independent biological replicates; viral yields decreased after RNF185 knockdown and increased after RNF185 overexpression | Chen 2024, *Autophagy*. https://doi.org/10.1080/15548627.2023.2293442 | (pqac-00000023, pqac-00000024, pqac-00000025, pqac-00000026, pqac-00000027) |
| Pathway / phenotype | CRISPR screens show RNF185 is a redundant ER-resident E3 for CFTR-F508del ERAD in parallel with RNF5; genetic data suggest UBE2D3 may act as an E2 for both RNF5 and RNF185 | Genome-wide CRISPR/Cas9 stability screen; sensitized sublibrary screens in RNF5KO and RNF5/UBE2D3KO backgrounds; double-knockout analysis and degradation kinetics | RNF5 was top E3 hit, but RNF5 KO only modestly reduced CFTR-F508del degradation; RNF185 emerged as a modest hit in sensitized screens; RNF185/UBE2D3KO kinetics were indistinguishable from UBE2D3KO, supporting same-pathway action; RNF5 disruption plus correctors increased mNG-F508del half-life and maturation by 52% and 46%, respectively; triple RNF185/RNF5/UBE2D3 knockout caused severe growth defect | Riepe 2024, *Molecular Biology of the Cell*. https://doi.org/10.1091/mbc.e23-08-0336 | (pqac-00000016, pqac-00000017, pqac-00000018, pqac-00000019) |
| Expert synthesis | Recent expert review places RNF185 at the intersection of mitochondrial quality control, ERAD, autophagy/mitophagy, innate immunity, and cancer biology; known targets/processes include BNIP1, CFTR, cGAS, and RNF185/Membralin-dependent ER quality control | Narrative synthesis of primary literature on mitochondrial E3 ligases and cancer/mitochondrial biology | Review emphasizes context-dependent tumor-suppressive versus oncogenic roles, epigenetic/miRNA regulation, and major knowledge gaps in substrate scope and cooperation with other E3 ligases | Di Gregorio 2023, *International Journal of Molecular Sciences*. https://doi.org/10.3390/ijms242417176 | (pqac-00000029, pqac-00000030, pqac-00000031, pqac-00000032, pqac-00000034) |


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