{"gene":"RNF185","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2011,"finding":"RNF185 localizes to the mitochondrial outer membrane via its two C-terminal transmembrane domains, where it acts as an E3 ubiquitin ligase that stimulates LC3II accumulation and autophagolysosome formation, promoting selective mitochondrial autophagy.","method":"Subcellular fractionation, live-cell imaging, co-immunoprecipitation, in vivo ubiquitination assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (fractionation, imaging, Co-IP, ubiquitination assay) from a single lab","pmids":["21931693"],"is_preprint":false},{"year":2011,"finding":"RNF185 directly ubiquitinates BNIP1 via K63-linked polyubiquitin chains; polyubiquitinated BNIP1 recruits the autophagy receptor p62, which binds both ubiquitin and LC3 to link ubiquitination with autophagy.","method":"Co-immunoprecipitation, in vivo ubiquitination assay with linkage-specific ubiquitin mutants","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus linkage-specific ubiquitination assay, single lab","pmids":["21931693"],"is_preprint":false},{"year":2013,"finding":"RNF185 controls the stability of CFTR and CFTRΔF508 in a RING domain- and proteasome-dependent manner, targeting CFTR to co-translational degradation as part of ERAD; RNF185 and RNF5 together form an E3 ligase module central to CFTR degradation.","method":"RNA interference (siRNA knockdown), turnover/pulse-chase analysis, proteasome inhibitor treatment, RING domain mutant","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (KD, turnover analysis, domain mutant, epistasis with RNF5), replicated with two CFTR substrates","pmids":["24019521"],"is_preprint":false},{"year":2014,"finding":"RNF185 interacts with Dvl2, a mediator of Wnt signaling, promotes its ubiquitination and proteasomal degradation, thereby inhibiting β-catenin-mediated transcriptional activity and negatively regulating osteogenic differentiation.","method":"Co-immunoprecipitation, ubiquitination assay, overexpression/knockdown with ALP activity and qRT-PCR readouts, rescue with Dvl2","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP, ubiquitination assay, and functional rescue, all from a single lab","pmids":["24727453"],"is_preprint":false},{"year":2017,"finding":"RNF185 (localized to the ER) interacts with cGAS and catalyzes K27-linked polyubiquitination of cGAS, promoting its enzymatic activity and enhancing downstream IRF3-responsive gene expression during HSV-1 infection.","method":"Co-immunoprecipitation, in vivo ubiquitination assay with linkage-specific ubiquitin mutants, ectopic expression and siRNA knockdown with reporter assays","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, linkage-specific ubiquitination, gain- and loss-of-function with defined signaling readout, single lab with multiple orthogonal methods","pmids":["28273161"],"is_preprint":false},{"year":2018,"finding":"RNF185 directly interacts with JWA and promotes its ubiquitination at lysine 158, leading to proteasomal degradation of JWA and facilitating gastric cancer cell migration and metastasis.","method":"Co-immunoprecipitation, ubiquitination assay with site-specific mutant (K158), overexpression/knockdown with migration and in vivo metastasis assays, rescue experiment","journal":"Biochimica et biophysica acta. Molecular basis of disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, site-specific mutagenesis of substrate, in vivo rescue, single lab","pmids":["29481911"],"is_preprint":false},{"year":2020,"finding":"RNF185 forms an ER membrane complex with TMUB1/2 and TMEM259/Membralin (an RNF185/Membralin complex) that cooperates with cytosolic ubiquitin ligase UBE3C and the p97 ATPase to degrade a subset of misfolded ER membrane proteins via ERAD.","method":"CRISPR-Cas9 genome-wide screen, biochemical fractionation, mass spectrometry, co-immunoprecipitation","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — unbiased genome-wide screen plus MS-defined complex plus biochemical validation, multiple orthogonal methods","pmids":["32738194"],"is_preprint":false},{"year":2022,"finding":"RNF185 polyubiquitinates Ebolavirus GP1,2 on lysine 673 via K27-linked ubiquitin chains; polyubiquitinated GP1,2 is subsequently recruited into autophagosomes by p62/SQSTM1 in an ATG3- and ATG5-dependent manner, directing GP1,2 to lysosomal (rather than proteasomal) degradation.","method":"Co-immunoprecipitation, in vivo ubiquitination assay with site-directed mutagenesis (K673), ATG3/ATG5 knockout analysis, autophagy flux assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — site-specific mutagenesis of substrate, multiple genetic KOs, autophagy flux assays, and linkage-specific ubiquitination in a single rigorous study","pmids":["36224200"],"is_preprint":false},{"year":2023,"finding":"RNF185 co-localizes with the SARS-CoV-2 envelope protein at the ER and regulates its stability; depletion of RNF185 significantly increases SARS-CoV-2 viral titer in a cellular model.","method":"Genetic screen, co-localization imaging, RNF185 siRNA knockdown with viral titer measurement","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — genetic screen plus imaging plus functional viral titer readout, single lab","pmids":["37095859"],"is_preprint":false},{"year":2024,"finding":"RNF185 interacts with TUFM via its transmembrane domain 1 and catalyzes K27-linked polyubiquitination of TUFM, enabling SQSTM1/p62 recognition and initiating mitophagy during Senecavirus A infection.","method":"Co-immunoprecipitation, GST pulldown, in vivo ubiquitination assay with linkage-specific ubiquitin mutants, site-directed mutagenesis of TUFM (E196, E211), domain deletion analysis","journal":"Autophagy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, GST pulldown, linkage-specific and site-specific ubiquitination, domain mutants, single lab","pmids":["38084826"],"is_preprint":false},{"year":2024,"finding":"The RNF185/Membralin ERAD complex recognizes unassembled Tapasin (a component of the MHC-I peptide loading complex) and targets it for degradation; loss of RNF185/Membralin elevates Tapasin steady-state levels and increases MHC-I surface expression on antigen-presenting cells.","method":"Unbiased proteomics screen, co-immunoprecipitation, CRISPR knockout, flow cytometry for MHC-I surface levels, degradation assays","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — unbiased proteomics screen, CRISPR KO, Co-IP, and functional surface MHC-I readout with multiple orthogonal methods","pmids":["39353943"],"is_preprint":false},{"year":2024,"finding":"D-mannose treatment upregulates RNF185 expression, which then interacts with and promotes proteasomal degradation of IDH2 via ubiquitination, inhibiting IDH2-mediated NADPH production in breast cancer cells.","method":"RNA-seq, Western blot, co-immunoprecipitation, ubiquitination assay, NADPH production assay, colony formation and CCK-8 assays","journal":"Nutrition & metabolism","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP and ubiquitination assay plus functional readout, single lab, single study","pmids":["38167476"],"is_preprint":false},{"year":2024,"finding":"RNF185 depletion in prostate cancer cells increases COL3A1 levels and promotes cell migration and metastasis; co-inhibition of COL3A1 rescues the enhanced migration phenotype, placing COL3A1 downstream of RNF185 in a metastasis-suppressive pathway.","method":"shRNA knockdown, RNA-sequencing, in vitro migration/invasion assays, in vivo xenograft mouse model, COL3A1 co-inhibition rescue","journal":"Molecular cancer research : MCR","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo metastasis model, RNA-seq, genetic epistasis via rescue, single lab","pmids":["37831068"],"is_preprint":false},{"year":2025,"finding":"RNF185 mediates ubiquitination of BAK1 to maintain mitochondrial integrity; loss of RNF185 causes BAK1 accumulation, mtDNA release, and activation of the cGAS-STING-IRF3 pathway, forming a positive feedback loop that promotes apoptosis in ESCC cells.","method":"CRISPR/Cas9 knockout, immunoprecipitation, ubiquitination assay, Western blot, luciferase reporter assay, ChIP-qPCR, transmission electron microscopy, xenograft in vivo model","journal":"European journal of medical research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP, ubiquitination assay, multiple pathway readouts, in vivo model, single lab","pmids":["41254727"],"is_preprint":false},{"year":2026,"finding":"Upon DNA damage, RNF185 undergoes ATM/ATR-dependent phosphorylation at threonine 106, translocates to the nucleus via NUP88 interaction, and sequentially promotes K6/K63-linked ubiquitination of RPA1 (stabilizing RPA1 on ssDNA for replication fork restart) followed by K48-linked ubiquitination at RPA1 lysine 458 (promoting RPA1 degradation and removal from chromatin for HR completion); RNF185 competes with deubiquitinase OTUB1 for RPA1 binding.","method":"Co-immunoprecipitation, in vivo ubiquitination assay with linkage-specific mutants and site-directed mutagenesis (T106, K458), ATM/ATR inhibitor treatment, OTUB1 competition assay, HR efficiency assay, replication fork stability assay, in vivo irradiation/cisplatin sensitivity","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (site-specific mutagenesis, linkage-specific ubiquitination, competition assay, functional HR and fork restart readouts), single lab, single study","pmids":["41997286"],"is_preprint":false}],"current_model":"RNF185 is a RING-domain E3 ubiquitin ligase that resides at the ER and mitochondrial outer membranes and operates through multiple substrate-specific mechanisms: it catalyzes K27-linked ubiquitination of cGAS to enhance innate immune signaling, K63-linked ubiquitination of BNIP1 and TUFM to drive selective mitophagy/autophagy via p62-LC3 recruitment, and ERAD-directed ubiquitination of misfolded ER membrane proteins (CFTR, EBOV-GP1,2, Tapasin) in a complex with Membralin/TMUB1/TMUB2 and co-operating with UBE3C and p97; additionally, RNF185 ubiquitinates cytoplasmic substrates including JWA, Dvl2, BAK1, and IDH2 for proteasomal degradation, and upon DNA damage undergoes ATM/ATR-dependent nuclear translocation to temporally switch RPA1 ubiquitin linkage (K6/K63 then K48), thereby coordinating replication fork restart and homologous recombination."},"narrative":{"mechanistic_narrative":"RNF185 is a RING-domain E3 ubiquitin ligase anchored at the ER and mitochondrial outer membranes that selects diverse substrates and writes distinct ubiquitin linkages to route them toward proteasomal degradation, selective autophagy, or signaling activation [PMID:21931693, PMID:24019521, PMID:28273161]. At the ER it forms a membrane-embedded ERAD module with TMUB1/2 and Membralin (TMEM259) that cooperates with the cytosolic ligase UBE3C and the p97 ATPase to degrade misfolded or unassembled membrane proteins, including CFTR and CFTRΔF508, unassembled Tapasin, and viral envelope glycoproteins, thereby controlling protein quality control and indirectly setting MHC-I surface levels [PMID:24019521, PMID:32738194, PMID:39353943]. Through K27- and K63-linked chains it functions as a positive regulator of innate immunity and selective autophagy: it ubiquitinates cGAS (K27) to enhance its enzymatic activity and IRF3-dependent gene expression [PMID:28273161], and modifies mitochondrial substrates BNIP1 (K63) and TUFM (K27) to recruit the autophagy receptor p62/SQSTM1 and drive LC3-dependent mitophagy [PMID:21931693, PMID:38084826]. RNF185 also degrades a range of cytoplasmic and signaling substrates — Dvl2 to restrain Wnt/β-catenin signaling [PMID:24727453], JWA to promote gastric cancer cell migration [PMID:29481911], IDH2 to limit NADPH production [PMID:38167476], and BAK1 to maintain mitochondrial integrity and restrain cGAS-STING-driven apoptosis [PMID:41254727]. Upon DNA damage it undergoes ATM/ATR-dependent phosphorylation, NUP88-mediated nuclear translocation, and sequential switching of RPA1 ubiquitin linkages (K6/K63 then K48), coordinating replication fork restart and homologous recombination in competition with the deubiquitinase OTUB1 [PMID:41997286].","teleology":[{"year":2011,"claim":"Established RNF185 as a mitochondrial outer-membrane E3 ligase that links substrate ubiquitination to selective autophagy, answering where the enzyme acts and how it engages the autophagy machinery.","evidence":"Subcellular fractionation, imaging, Co-IP and in vivo ubiquitination assays showing K63-linked ubiquitination of BNIP1 and p62 recruitment","pmids":["21931693"],"confidence":"Medium","gaps":["Single lab; physiological trigger for mitophagy not defined","Whether BNIP1 is the principal mitophagy substrate not established"]},{"year":2013,"claim":"Defined RNF185 as a core ERAD ligase by showing it controls CFTR turnover in a RING- and proteasome-dependent manner together with RNF5, establishing its protein quality-control role.","evidence":"siRNA knockdown, pulse-chase turnover, proteasome inhibition, RING mutant, epistasis with RNF5 in two CFTR substrates","pmids":["24019521"],"confidence":"High","gaps":["Mechanism of misfolded-substrate recognition not resolved","Ubiquitin linkage type on CFTR not specified"]},{"year":2014,"claim":"Extended RNF185 substrate range to cytoplasmic signaling by showing it degrades Dvl2 to suppress Wnt/β-catenin output and osteogenic differentiation.","evidence":"Co-IP, ubiquitination assay, gain/loss-of-function with ALP and qRT-PCR readouts, Dvl2 rescue","pmids":["24727453"],"confidence":"Medium","gaps":["Ubiquitin linkage on Dvl2 not defined","Single lab; in vivo relevance untested"]},{"year":2017,"claim":"Showed RNF185 can be an activating rather than degradative ligase, using K27-linked ubiquitination of cGAS to potentiate innate antiviral signaling.","evidence":"Reciprocal Co-IP, linkage-specific ubiquitination, gain/loss-of-function reporter assays during HSV-1 infection","pmids":["28273161"],"confidence":"High","gaps":["CGAS ubiquitination site not mapped","Structural basis for non-degradative K27 chains unknown"]},{"year":2018,"claim":"Identified site-specific (K158) ubiquitination of JWA by RNF185 as a pro-metastatic axis in gastric cancer, linking the ligase to tumor cell migration.","evidence":"Co-IP, site-specific mutagenesis, migration/in vivo metastasis assays, rescue","pmids":["29481911"],"confidence":"Medium","gaps":["Single lab; broader substrate context unclear","Regulation of RNF185 in tumors not defined"]},{"year":2020,"claim":"Resolved the molecular architecture of RNF185-dependent ERAD by defining the RNF185/Membralin/TMUB1/2 complex acting with UBE3C and p97 on a specific subset of misfolded membrane proteins.","evidence":"Genome-wide CRISPR screen, biochemical fractionation, mass spectrometry, Co-IP","pmids":["32738194"],"confidence":"High","gaps":["Substrate selectivity determinants within the complex not fully defined","Stoichiometry of the membrane complex not resolved"]},{"year":2022,"claim":"Demonstrated RNF185 can route a substrate to autophagy rather than the proteasome, K27-ubiquitinating Ebolavirus GP1,2 at K673 for p62/ATG-dependent lysosomal degradation.","evidence":"Co-IP, site-directed mutagenesis (K673), ATG3/ATG5 knockouts, autophagy flux assays, linkage-specific ubiquitination","pmids":["36224200"],"confidence":"High","gaps":["What dictates lysosomal vs proteasomal fate of K27-modified substrates unknown"]},{"year":2023,"claim":"Implicated RNF185 in restricting SARS-CoV-2 by regulating envelope protein stability at the ER, expanding its antiviral role.","evidence":"Genetic screen, co-localization imaging, siRNA knockdown with viral titer readout","pmids":["37095859"],"confidence":"Medium","gaps":["Direct ubiquitination of E protein not demonstrated","Linkage type and degradation route unspecified"]},{"year":2024,"claim":"Connected RNF185-dependent ERAD to immune presentation by showing the RNF185/Membralin complex degrades unassembled Tapasin and thereby limits MHC-I surface expression.","evidence":"Unbiased proteomics, CRISPR knockout, Co-IP, flow cytometry for surface MHC-I, degradation assays","pmids":["39353943"],"confidence":"High","gaps":["Whether Tapasin degradation is the dominant determinant of MHC-I changes in vivo not established"]},{"year":2024,"claim":"Established TUFM as a mitochondrial RNF185 substrate, with K27-linked ubiquitination via transmembrane domain 1 enabling p62-driven mitophagy during Senecavirus A infection.","evidence":"Co-IP, GST pulldown, linkage- and site-specific ubiquitination, domain deletion analysis","pmids":["38084826"],"confidence":"Medium","gaps":["Generality of TUFM mitophagy beyond viral infection unclear","Single lab"]},{"year":2024,"claim":"Linked RNF185 to metabolic control, showing D-mannose-induced RNF185 degrades IDH2 to suppress NADPH production in breast cancer cells.","evidence":"RNA-seq, Co-IP, ubiquitination and NADPH assays, colony formation/CCK-8 readouts","pmids":["38167476"],"confidence":"Medium","gaps":["Mechanism of D-mannose-driven RNF185 upregulation not defined","Ubiquitin linkage on IDH2 unspecified"]},{"year":2024,"claim":"Defined a metastasis-suppressive RNF185 pathway in prostate cancer acting upstream of COL3A1.","evidence":"shRNA knockdown, RNA-seq, migration/invasion assays, xenograft model, COL3A1 co-inhibition rescue","pmids":["37831068"],"confidence":"Medium","gaps":["Whether COL3A1 regulation is direct or transcriptional not resolved","No direct ubiquitination substrate identified in this axis"]},{"year":2025,"claim":"Showed RNF185 ubiquitinates BAK1 to preserve mitochondrial integrity, with its loss triggering mtDNA release and cGAS-STING-IRF3-driven apoptosis in ESCC.","evidence":"CRISPR knockout, IP, ubiquitination assay, luciferase reporter, ChIP-qPCR, electron microscopy, xenograft","pmids":["41254727"],"confidence":"Medium","gaps":["BAK1 ubiquitination site and linkage not defined","Relationship to RNF185's cGAS-activating role not reconciled"]},{"year":2026,"claim":"Revealed a damage-induced nuclear function in which phosphorylated RNF185 sequentially switches RPA1 ubiquitin linkages to coordinate fork restart and homologous recombination.","evidence":"Co-IP, linkage- and site-specific ubiquitination (T106, K458), ATM/ATR inhibition, OTUB1 competition, HR and fork-restart assays, in vivo irradiation/cisplatin sensitivity","pmids":["41997286"],"confidence":"Medium","gaps":["Single lab; nuclear translocation kinetics relative to membrane pool unclear","How the K6/K63-to-K48 switch is timed not mechanistically resolved"]},{"year":null,"claim":"How RNF185 selects among its many substrates and partitions between ER membrane, mitochondrial, and nuclear pools to specify proteasomal degradation, autophagy, or signaling activation remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying determinant of substrate or linkage choice identified","Dynamic regulation of subcellular distribution not characterized","No structural model of RNF185 substrate engagement"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,1,2,4,6,7,9,14]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[2,4,5,7,9,11,13,14]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[2,6,14]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[4,6,8,10]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[0,9,13]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[14]}],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[2,6,10]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[0,1,7,9]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[4,10,13]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[14]}],"complexes":["RNF185/Membralin ERAD complex (with TMUB1/TMUB2)","RNF185-RNF5 ERAD module"],"partners":["RNF5","TMEM259","TMUB1","TMUB2","UBE3C","CGAS","TUFM","RPA1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q96GF1","full_name":"E3 ubiquitin-protein ligase RNF185","aliases":["RING finger protein 185"],"length_aa":192,"mass_kda":20.5,"function":"E3 ubiquitin-protein ligase that regulates selective mitochondrial autophagy by mediating 'Lys-63'-linked polyubiquitination of BNIP1 (PubMed:21931693). Acts in the endoplasmic reticulum (ER)-associated degradation (ERAD) pathway, which targets misfolded proteins that accumulate in the endoplasmic reticulum (ER) for ubiquitination and subsequent proteasome-mediated degradation (PubMed:27485036). Protects cells from ER stress-induced apoptosis (PubMed:27485036). Responsible for the cotranslational ubiquitination and degradation of CFTR in the ERAD pathway (PubMed:24019521). Also acts as a regulator of the innate antiviral response by catalyzing 'Lys-27'-linked polyubiquitination of CGAS at 'Lys-173' and 'Lys-384', thereby promoting CGAS cyclic GMP-AMP synthase activity (PubMed:28273161). Preferentially associates with the E2 enzymes UBE2J1 and UBE2J2 (PubMed:24019521)","subcellular_location":"Mitochondrion outer membrane; Endoplasmic reticulum membrane","url":"https://www.uniprot.org/uniprotkb/Q96GF1/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF185","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"CANX","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/RNF185","total_profiled":1310},"omim":[{"mim_id":"620096","title":"RING FINGER PROTEIN 185; RNF185","url":"https://www.omim.org/entry/620096"},{"mim_id":"613297","title":"MEMBRANE-ASSOCIATED RING-CH FINGER PROTEIN 6; MARCHF6","url":"https://www.omim.org/entry/613297"},{"mim_id":"606130","title":"RING FINGER PROTEIN 26; RNF26","url":"https://www.omim.org/entry/606130"},{"mim_id":"603291","title":"BCL2/ADENOVIRUS E1B 19-KD PROTEIN-INTERACTING PROTEIN 1; BNIP1","url":"https://www.omim.org/entry/603291"},{"mim_id":"602421","title":"CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR; CFTR","url":"https://www.omim.org/entry/602421"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RNF185"},"hgnc":{"alias_symbol":["FLJ38628"],"prev_symbol":[]},"alphafold":{"accession":"Q96GF1","domains":[{"cath_id":"3.30.40.10","chopping":"35-89","consensus_level":"medium","plddt":85.934,"start":35,"end":89}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96GF1","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q96GF1-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q96GF1-F1-predicted_aligned_error_v6.png","plddt_mean":68.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF185","jax_strain_url":"https://www.jax.org/strain/search?query=RNF185"},"sequence":{"accession":"Q96GF1","fasta_url":"https://rest.uniprot.org/uniprotkb/Q96GF1.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q96GF1/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96GF1"}},"corpus_meta":[{"pmid":"28273161","id":"PMC_28273161","title":"The E3 ubiquitin ligase RNF185 facilitates the cGAS-mediated innate immune response.","date":"2017","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/28273161","citation_count":146,"is_preprint":false},{"pmid":"21931693","id":"PMC_21931693","title":"RNF185, a novel mitochondrial ubiquitin E3 ligase, regulates autophagy through interaction with BNIP1.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21931693","citation_count":86,"is_preprint":false},{"pmid":"24019521","id":"PMC_24019521","title":"RNF185 is a novel E3 ligase of endoplasmic reticulum-associated degradation (ERAD) that targets cystic fibrosis transmembrane conductance regulator (CFTR).","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24019521","citation_count":83,"is_preprint":false},{"pmid":"32738194","id":"PMC_32738194","title":"Quality Control of ER Membrane Proteins by the RNF185/Membralin Ubiquitin Ligase Complex.","date":"2020","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/32738194","citation_count":53,"is_preprint":false},{"pmid":"29481911","id":"PMC_29481911","title":"RNF185 modulates JWA ubiquitination and promotes gastric cancer metastasis.","date":"2018","source":"Biochimica et biophysica acta. 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research","url":"https://pubmed.ncbi.nlm.nih.gov/41254727","citation_count":3,"is_preprint":false},{"pmid":"38167476","id":"PMC_38167476","title":"D-mannose promotes the degradation of IDH2 through upregulation of RNF185 and suppresses breast cancer.","date":"2024","source":"Nutrition & metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/38167476","citation_count":3,"is_preprint":false},{"pmid":"41997286","id":"PMC_41997286","title":"RNF185 orchestrates replication fork restart and homologous recombination through temporal RPA1 ubiquitination switching.","date":"2026","source":"Cancer letters","url":"https://pubmed.ncbi.nlm.nih.gov/41997286","citation_count":0,"is_preprint":false},{"pmid":"37425866","id":"PMC_37425866","title":"RNF185 control of COL3A1 expression limits prostate cancer migration and metastatic potential.","date":"2023","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/37425866","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11076,"output_tokens":3767,"usd":0.044866,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11366,"output_tokens":4825,"usd":0.088728,"stage2_stop_reason":"end_turn"},"total_usd":0.133594,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2011,\n      \"finding\": \"RNF185 localizes to the mitochondrial outer membrane via its two C-terminal transmembrane domains, where it acts as an E3 ubiquitin ligase that stimulates LC3II accumulation and autophagolysosome formation, promoting selective mitochondrial autophagy.\",\n      \"method\": \"Subcellular fractionation, live-cell imaging, co-immunoprecipitation, in vivo ubiquitination assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (fractionation, imaging, Co-IP, ubiquitination assay) from a single lab\",\n      \"pmids\": [\"21931693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"RNF185 directly ubiquitinates BNIP1 via K63-linked polyubiquitin chains; polyubiquitinated BNIP1 recruits the autophagy receptor p62, which binds both ubiquitin and LC3 to link ubiquitination with autophagy.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay with linkage-specific ubiquitin mutants\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus linkage-specific ubiquitination assay, single lab\",\n      \"pmids\": [\"21931693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RNF185 controls the stability of CFTR and CFTRΔF508 in a RING domain- and proteasome-dependent manner, targeting CFTR to co-translational degradation as part of ERAD; RNF185 and RNF5 together form an E3 ligase module central to CFTR degradation.\",\n      \"method\": \"RNA interference (siRNA knockdown), turnover/pulse-chase analysis, proteasome inhibitor treatment, RING domain mutant\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (KD, turnover analysis, domain mutant, epistasis with RNF5), replicated with two CFTR substrates\",\n      \"pmids\": [\"24019521\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RNF185 interacts with Dvl2, a mediator of Wnt signaling, promotes its ubiquitination and proteasomal degradation, thereby inhibiting β-catenin-mediated transcriptional activity and negatively regulating osteogenic differentiation.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, overexpression/knockdown with ALP activity and qRT-PCR readouts, rescue with Dvl2\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP, ubiquitination assay, and functional rescue, all from a single lab\",\n      \"pmids\": [\"24727453\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RNF185 (localized to the ER) interacts with cGAS and catalyzes K27-linked polyubiquitination of cGAS, promoting its enzymatic activity and enhancing downstream IRF3-responsive gene expression during HSV-1 infection.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay with linkage-specific ubiquitin mutants, ectopic expression and siRNA knockdown with reporter assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, linkage-specific ubiquitination, gain- and loss-of-function with defined signaling readout, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"28273161\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RNF185 directly interacts with JWA and promotes its ubiquitination at lysine 158, leading to proteasomal degradation of JWA and facilitating gastric cancer cell migration and metastasis.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay with site-specific mutant (K158), overexpression/knockdown with migration and in vivo metastasis assays, rescue experiment\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular basis of disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, site-specific mutagenesis of substrate, in vivo rescue, single lab\",\n      \"pmids\": [\"29481911\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF185 forms an ER membrane complex with TMUB1/2 and TMEM259/Membralin (an RNF185/Membralin complex) that cooperates with cytosolic ubiquitin ligase UBE3C and the p97 ATPase to degrade a subset of misfolded ER membrane proteins via ERAD.\",\n      \"method\": \"CRISPR-Cas9 genome-wide screen, biochemical fractionation, mass spectrometry, co-immunoprecipitation\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — unbiased genome-wide screen plus MS-defined complex plus biochemical validation, multiple orthogonal methods\",\n      \"pmids\": [\"32738194\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF185 polyubiquitinates Ebolavirus GP1,2 on lysine 673 via K27-linked ubiquitin chains; polyubiquitinated GP1,2 is subsequently recruited into autophagosomes by p62/SQSTM1 in an ATG3- and ATG5-dependent manner, directing GP1,2 to lysosomal (rather than proteasomal) degradation.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay with site-directed mutagenesis (K673), ATG3/ATG5 knockout analysis, autophagy flux assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — site-specific mutagenesis of substrate, multiple genetic KOs, autophagy flux assays, and linkage-specific ubiquitination in a single rigorous study\",\n      \"pmids\": [\"36224200\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF185 co-localizes with the SARS-CoV-2 envelope protein at the ER and regulates its stability; depletion of RNF185 significantly increases SARS-CoV-2 viral titer in a cellular model.\",\n      \"method\": \"Genetic screen, co-localization imaging, RNF185 siRNA knockdown with viral titer measurement\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — genetic screen plus imaging plus functional viral titer readout, single lab\",\n      \"pmids\": [\"37095859\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF185 interacts with TUFM via its transmembrane domain 1 and catalyzes K27-linked polyubiquitination of TUFM, enabling SQSTM1/p62 recognition and initiating mitophagy during Senecavirus A infection.\",\n      \"method\": \"Co-immunoprecipitation, GST pulldown, in vivo ubiquitination assay with linkage-specific ubiquitin mutants, site-directed mutagenesis of TUFM (E196, E211), domain deletion analysis\",\n      \"journal\": \"Autophagy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, GST pulldown, linkage-specific and site-specific ubiquitination, domain mutants, single lab\",\n      \"pmids\": [\"38084826\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"The RNF185/Membralin ERAD complex recognizes unassembled Tapasin (a component of the MHC-I peptide loading complex) and targets it for degradation; loss of RNF185/Membralin elevates Tapasin steady-state levels and increases MHC-I surface expression on antigen-presenting cells.\",\n      \"method\": \"Unbiased proteomics screen, co-immunoprecipitation, CRISPR knockout, flow cytometry for MHC-I surface levels, degradation assays\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — unbiased proteomics screen, CRISPR KO, Co-IP, and functional surface MHC-I readout with multiple orthogonal methods\",\n      \"pmids\": [\"39353943\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"D-mannose treatment upregulates RNF185 expression, which then interacts with and promotes proteasomal degradation of IDH2 via ubiquitination, inhibiting IDH2-mediated NADPH production in breast cancer cells.\",\n      \"method\": \"RNA-seq, Western blot, co-immunoprecipitation, ubiquitination assay, NADPH production assay, colony formation and CCK-8 assays\",\n      \"journal\": \"Nutrition & metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP and ubiquitination assay plus functional readout, single lab, single study\",\n      \"pmids\": [\"38167476\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF185 depletion in prostate cancer cells increases COL3A1 levels and promotes cell migration and metastasis; co-inhibition of COL3A1 rescues the enhanced migration phenotype, placing COL3A1 downstream of RNF185 in a metastasis-suppressive pathway.\",\n      \"method\": \"shRNA knockdown, RNA-sequencing, in vitro migration/invasion assays, in vivo xenograft mouse model, COL3A1 co-inhibition rescue\",\n      \"journal\": \"Molecular cancer research : MCR\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo metastasis model, RNA-seq, genetic epistasis via rescue, single lab\",\n      \"pmids\": [\"37831068\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF185 mediates ubiquitination of BAK1 to maintain mitochondrial integrity; loss of RNF185 causes BAK1 accumulation, mtDNA release, and activation of the cGAS-STING-IRF3 pathway, forming a positive feedback loop that promotes apoptosis in ESCC cells.\",\n      \"method\": \"CRISPR/Cas9 knockout, immunoprecipitation, ubiquitination assay, Western blot, luciferase reporter assay, ChIP-qPCR, transmission electron microscopy, xenograft in vivo model\",\n      \"journal\": \"European journal of medical research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP, ubiquitination assay, multiple pathway readouts, in vivo model, single lab\",\n      \"pmids\": [\"41254727\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"Upon DNA damage, RNF185 undergoes ATM/ATR-dependent phosphorylation at threonine 106, translocates to the nucleus via NUP88 interaction, and sequentially promotes K6/K63-linked ubiquitination of RPA1 (stabilizing RPA1 on ssDNA for replication fork restart) followed by K48-linked ubiquitination at RPA1 lysine 458 (promoting RPA1 degradation and removal from chromatin for HR completion); RNF185 competes with deubiquitinase OTUB1 for RPA1 binding.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay with linkage-specific mutants and site-directed mutagenesis (T106, K458), ATM/ATR inhibitor treatment, OTUB1 competition assay, HR efficiency assay, replication fork stability assay, in vivo irradiation/cisplatin sensitivity\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (site-specific mutagenesis, linkage-specific ubiquitination, competition assay, functional HR and fork restart readouts), single lab, single study\",\n      \"pmids\": [\"41997286\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF185 is a RING-domain E3 ubiquitin ligase that resides at the ER and mitochondrial outer membranes and operates through multiple substrate-specific mechanisms: it catalyzes K27-linked ubiquitination of cGAS to enhance innate immune signaling, K63-linked ubiquitination of BNIP1 and TUFM to drive selective mitophagy/autophagy via p62-LC3 recruitment, and ERAD-directed ubiquitination of misfolded ER membrane proteins (CFTR, EBOV-GP1,2, Tapasin) in a complex with Membralin/TMUB1/TMUB2 and co-operating with UBE3C and p97; additionally, RNF185 ubiquitinates cytoplasmic substrates including JWA, Dvl2, BAK1, and IDH2 for proteasomal degradation, and upon DNA damage undergoes ATM/ATR-dependent nuclear translocation to temporally switch RPA1 ubiquitin linkage (K6/K63 then K48), thereby coordinating replication fork restart and homologous recombination.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RNF185 is a RING-domain E3 ubiquitin ligase anchored at the ER and mitochondrial outer membranes that selects diverse substrates and writes distinct ubiquitin linkages to route them toward proteasomal degradation, selective autophagy, or signaling activation [#0, #2, #4]. At the ER it forms a membrane-embedded ERAD module with TMUB1/2 and Membralin (TMEM259) that cooperates with the cytosolic ligase UBE3C and the p97 ATPase to degrade misfolded or unassembled membrane proteins, including CFTR and CFTR\\u0394F508, unassembled Tapasin, and viral envelope glycoproteins, thereby controlling protein quality control and indirectly setting MHC-I surface levels [#2, #6, #10]. Through K27- and K63-linked chains it functions as a positive regulator of innate immunity and selective autophagy: it ubiquitinates cGAS (K27) to enhance its enzymatic activity and IRF3-dependent gene expression [#4], and modifies mitochondrial substrates BNIP1 (K63) and TUFM (K27) to recruit the autophagy receptor p62/SQSTM1 and drive LC3-dependent mitophagy [#1, #9]. RNF185 also degrades a range of cytoplasmic and signaling substrates \\u2014 Dvl2 to restrain Wnt/\\u03b2-catenin signaling [#3], JWA to promote gastric cancer cell migration [#5], IDH2 to limit NADPH production [#11], and BAK1 to maintain mitochondrial integrity and restrain cGAS-STING-driven apoptosis [#13]. Upon DNA damage it undergoes ATM/ATR-dependent phosphorylation, NUP88-mediated nuclear translocation, and sequential switching of RPA1 ubiquitin linkages (K6/K63 then K48), coordinating replication fork restart and homologous recombination in competition with the deubiquitinase OTUB1 [#14].\",\n  \"teleology\": [\n    {\n      \"year\": 2011,\n      \"claim\": \"Established RNF185 as a mitochondrial outer-membrane E3 ligase that links substrate ubiquitination to selective autophagy, answering where the enzyme acts and how it engages the autophagy machinery.\",\n      \"evidence\": \"Subcellular fractionation, imaging, Co-IP and in vivo ubiquitination assays showing K63-linked ubiquitination of BNIP1 and p62 recruitment\",\n      \"pmids\": [\"21931693\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single lab; physiological trigger for mitophagy not defined\", \"Whether BNIP1 is the principal mitophagy substrate not established\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defined RNF185 as a core ERAD ligase by showing it controls CFTR turnover in a RING- and proteasome-dependent manner together with RNF5, establishing its protein quality-control role.\",\n      \"evidence\": \"siRNA knockdown, pulse-chase turnover, proteasome inhibition, RING mutant, epistasis with RNF5 in two CFTR substrates\",\n      \"pmids\": [\"24019521\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism of misfolded-substrate recognition not resolved\", \"Ubiquitin linkage type on CFTR not specified\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Extended RNF185 substrate range to cytoplasmic signaling by showing it degrades Dvl2 to suppress Wnt/\\u03b2-catenin output and osteogenic differentiation.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, gain/loss-of-function with ALP and qRT-PCR readouts, Dvl2 rescue\",\n      \"pmids\": [\"24727453\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Ubiquitin linkage on Dvl2 not defined\", \"Single lab; in vivo relevance untested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showed RNF185 can be an activating rather than degradative ligase, using K27-linked ubiquitination of cGAS to potentiate innate antiviral signaling.\",\n      \"evidence\": \"Reciprocal Co-IP, linkage-specific ubiquitination, gain/loss-of-function reporter assays during HSV-1 infection\",\n      \"pmids\": [\"28273161\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"CGAS ubiquitination site not mapped\", \"Structural basis for non-degradative K27 chains unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identified site-specific (K158) ubiquitination of JWA by RNF185 as a pro-metastatic axis in gastric cancer, linking the ligase to tumor cell migration.\",\n      \"evidence\": \"Co-IP, site-specific mutagenesis, migration/in vivo metastasis assays, rescue\",\n      \"pmids\": [\"29481911\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single lab; broader substrate context unclear\", \"Regulation of RNF185 in tumors not defined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Resolved the molecular architecture of RNF185-dependent ERAD by defining the RNF185/Membralin/TMUB1/2 complex acting with UBE3C and p97 on a specific subset of misfolded membrane proteins.\",\n      \"evidence\": \"Genome-wide CRISPR screen, biochemical fractionation, mass spectrometry, Co-IP\",\n      \"pmids\": [\"32738194\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Substrate selectivity determinants within the complex not fully defined\", \"Stoichiometry of the membrane complex not resolved\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrated RNF185 can route a substrate to autophagy rather than the proteasome, K27-ubiquitinating Ebolavirus GP1,2 at K673 for p62/ATG-dependent lysosomal degradation.\",\n      \"evidence\": \"Co-IP, site-directed mutagenesis (K673), ATG3/ATG5 knockouts, autophagy flux assays, linkage-specific ubiquitination\",\n      \"pmids\": [\"36224200\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"What dictates lysosomal vs proteasomal fate of K27-modified substrates unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Implicated RNF185 in restricting SARS-CoV-2 by regulating envelope protein stability at the ER, expanding its antiviral role.\",\n      \"evidence\": \"Genetic screen, co-localization imaging, siRNA knockdown with viral titer readout\",\n      \"pmids\": [\"37095859\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct ubiquitination of E protein not demonstrated\", \"Linkage type and degradation route unspecified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Connected RNF185-dependent ERAD to immune presentation by showing the RNF185/Membralin complex degrades unassembled Tapasin and thereby limits MHC-I surface expression.\",\n      \"evidence\": \"Unbiased proteomics, CRISPR knockout, Co-IP, flow cytometry for surface MHC-I, degradation assays\",\n      \"pmids\": [\"39353943\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether Tapasin degradation is the dominant determinant of MHC-I changes in vivo not established\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established TUFM as a mitochondrial RNF185 substrate, with K27-linked ubiquitination via transmembrane domain 1 enabling p62-driven mitophagy during Senecavirus A infection.\",\n      \"evidence\": \"Co-IP, GST pulldown, linkage- and site-specific ubiquitination, domain deletion analysis\",\n      \"pmids\": [\"38084826\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Generality of TUFM mitophagy beyond viral infection unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linked RNF185 to metabolic control, showing D-mannose-induced RNF185 degrades IDH2 to suppress NADPH production in breast cancer cells.\",\n      \"evidence\": \"RNA-seq, Co-IP, ubiquitination and NADPH assays, colony formation/CCK-8 readouts\",\n      \"pmids\": [\"38167476\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanism of D-mannose-driven RNF185 upregulation not defined\", \"Ubiquitin linkage on IDH2 unspecified\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a metastasis-suppressive RNF185 pathway in prostate cancer acting upstream of COL3A1.\",\n      \"evidence\": \"shRNA knockdown, RNA-seq, migration/invasion assays, xenograft model, COL3A1 co-inhibition rescue\",\n      \"pmids\": [\"37831068\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether COL3A1 regulation is direct or transcriptional not resolved\", \"No direct ubiquitination substrate identified in this axis\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Showed RNF185 ubiquitinates BAK1 to preserve mitochondrial integrity, with its loss triggering mtDNA release and cGAS-STING-IRF3-driven apoptosis in ESCC.\",\n      \"evidence\": \"CRISPR knockout, IP, ubiquitination assay, luciferase reporter, ChIP-qPCR, electron microscopy, xenograft\",\n      \"pmids\": [\"41254727\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"BAK1 ubiquitination site and linkage not defined\", \"Relationship to RNF185's cGAS-activating role not reconciled\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Revealed a damage-induced nuclear function in which phosphorylated RNF185 sequentially switches RPA1 ubiquitin linkages to coordinate fork restart and homologous recombination.\",\n      \"evidence\": \"Co-IP, linkage- and site-specific ubiquitination (T106, K458), ATM/ATR inhibition, OTUB1 competition, HR and fork-restart assays, in vivo irradiation/cisplatin sensitivity\",\n      \"pmids\": [\"41997286\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Single lab; nuclear translocation kinetics relative to membrane pool unclear\", \"How the K6/K63-to-K48 switch is timed not mechanistically resolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RNF185 selects among its many substrates and partitions between ER membrane, mitochondrial, and nuclear pools to specify proteasomal degradation, autophagy, or signaling activation remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No unifying determinant of substrate or linkage choice identified\", \"Dynamic regulation of subcellular distribution not characterized\", \"No structural model of RNF185 substrate engagement\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 1, 2, 4, 6, 7, 9, 14]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [2, 4, 5, 7, 9, 11, 13, 14]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [2, 6, 14]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [4, 6, 8, 10]},\n      {\"term_id\": \"GO:0005741\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [0, 9, 13]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [14]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 6, 10]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [0, 1, 7, 9]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [4, 10, 13]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [14]}\n    ],\n    \"complexes\": [\n      \"RNF185/Membralin ERAD complex (with TMUB1/TMUB2)\",\n      \"RNF185-RNF5 ERAD module\"\n    ],\n    \"partners\": [\n      \"RNF5\",\n      \"TMEM259\",\n      \"TMUB1\",\n      \"TMUB2\",\n      \"UBE3C\",\n      \"cGAS\",\n      \"TUFM\",\n      \"RPA1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}