{"gene":"RNF182","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":2008,"finding":"RNF182 is a brain-enriched E3 ubiquitin ligase that possesses RING finger-dependent E3 ubiquitin ligase activity, stimulating E2-dependent polyubiquitination in vitro, and directly interacts with ATP6V0C (a component of the V-ATPase involved in neurotransmitter release) to target it for proteasomal degradation.","method":"In vitro ubiquitination assay, yeast two-hybrid screening, co-precipitation (Co-IP), overexpression in cultured cells","journal":"Molecular neurodegeneration","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro enzymatic assay plus yeast two-hybrid and Co-IP orthogonal validation; foundational mechanistic paper","pmids":["18298843"],"is_preprint":false},{"year":2022,"finding":"RNF182 mediates K48-linked ubiquitination of p65 (NF-κB subunit), promoting its proteasomal degradation, thereby blocking p65 binding to the SLC7A11 promoter; this mechanism is activated downstream of PCDHB14 to induce ferroptosis in hepatocellular carcinoma cells.","method":"Co-immunoprecipitation, ubiquitination assay, chromatin immunoprecipitation, western blot, cell viability assays","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 — reciprocal Co-IP, ubiquitination assay, ChIP, and functional rescue experiments across multiple orthogonal methods","pmids":["35688944"],"is_preprint":false},{"year":2023,"finding":"RNF182 promotes ubiquitination and proteasomal degradation of p65 in lung adenocarcinoma (LUAD) cells, thereby suppressing p65-driven transcription of PDL1 and reducing immune evasion; RNF182 overexpression increased CD8+ T cell cytotoxicity against LUAD cells in co-culture.","method":"Co-immunoprecipitation, western blot, chromatin immunoprecipitation-qPCR, luciferase reporter assay, co-culture cytotoxicity assay, in vivo xenograft","journal":"Immunity, inflammation and disease","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal methods (Co-IP, ChIP, luciferase, in vivo) in a single study","pmids":["37249301"],"is_preprint":false},{"year":2022,"finding":"RNF182 facilitates K48-linked ubiquitination of p65, promoting its autophagic (p62-mediated) degradation; the compound Friedelin recruits RNF182 to increase this ubiquitination, thereby inhibiting NF-κB signaling in tenocytes and alleviating tendinopathy.","method":"Co-immunoprecipitation, ubiquitination assay (K48-linkage), western blot, blockade of ubiquitination as rescue experiment, in vivo mouse tendinopathy model","journal":"Nutrients","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP and ubiquitination assay with functional rescue, single lab","pmids":["35458235"],"is_preprint":false},{"year":2026,"finding":"RNF182 mediates ubiquitination and degradation of MFN2 (Mitofusin-2), leading to mitochondrial dysfunction; IL-1 receptor antagonist (rhIL-1Ra) suppresses TGF-β1-induced RNF182 expression, stabilizing MFN2 and reducing renal fibrosis.","method":"In vivo mouse models (UUO, 5/6Nx), in vitro TGF-β1-stimulated kidney cells, RNF182 inhibition rescue experiments, functional measurements of mitochondrial respiration and ATP production","journal":"Cell death discovery","confidence":"Medium","confidence_rationale":"Tier 2 — genetic/pharmacological loss-of-function with defined molecular and functional phenotype, rescue experiments; single lab","pmids":["41461638"],"is_preprint":false},{"year":2018,"finding":"RNF182 silencing in a rat myocardial ischemia-reperfusion injury model activates the mTOR signaling pathway (upregulating mTOR, S6K1, eEF2, Bcl-2) and reduces cardiomyocyte apoptosis and ventricular remodeling; these protective effects are reversed by mTOR inhibition (PITE), placing RNF182 upstream of mTOR as a negative regulator.","method":"shRNA knockdown in rat MIRI model, pharmacological mTOR inhibition, cardiac function measurements, apoptosis assays, protein expression analysis","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis via pharmacological rescue in vivo, single lab, no direct substrate identification","pmids":["30450663"],"is_preprint":false},{"year":2026,"finding":"RNF182 enhances p65 association with the autophagy receptor p62 and promotes autophagic degradation of p65 in nucleus pulposus cells; Friedelin treatment increases RNF182-p65 interaction and triggers selective p65 autophagy independently of IKK activity, inhibiting NF-κB signaling in intervertebral disc degeneration.","method":"In vivo mouse cervical spine instability model, in vitro nucleus pulposus cell assays, Co-immunoprecipitation, western blot, autophagy flux assays","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 — Co-IP and functional in vivo/in vitro experiments, single lab","pmids":["41503854"],"is_preprint":false},{"year":2010,"finding":"RNF182 expression is regulated by the transcriptional modulator MeCP2; chromatin immunoprecipitation confirmed MeCP2 binding to the methylated CpG island of the RNF182 gene, identifying RNF182 as a direct MeCP2 target gene involved in neuronal cell survival.","method":"Chromatin immunoprecipitation (ChIP), bisulfite sequencing, microarray expression profiling in MeCP2 mutant vs. wild-type clonal fibroblast cultures","journal":"Journal of cellular and molecular medicine","confidence":"Medium","confidence_rationale":"Tier 2 — ChIP with bisulfite validation; single lab, moderate method depth","pmids":["20569274"],"is_preprint":false},{"year":2023,"finding":"Benzo[a]pyrene (BaP) suppresses RNF182 expression in NSCLC cells via two mechanisms: AhR-dependent transcriptional repression and AhR-independent promoter hypermethylation; loss of RNF182 promotes cell proliferation and cell cycle progression in NSCLC cell lines.","method":"RT-qPCR, western blot, ChIP assay, bisulfite genomic sequencing (BGS), RNF182 overexpression cell assays","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods (ChIP, BGS, functional assays) in single lab","pmids":["36686776"],"is_preprint":false},{"year":2025,"finding":"Fish RNF182 (CiE3RNF182) associates with RIG-I and catalyzes Lys-33-linked polyubiquitination at the Lys33 residue of RIG-I, triggering RIG-I degradation and thereby inhibiting type I IFN antiviral signaling; CRISPR/Cas9 knockout of RNF182 in rare minnow enhanced survival against grass carp reovirus infection.","method":"Co-immunoprecipitation, ubiquitination assay (linkage-type analysis), CRISPR/Cas9 knockout, in vivo virus challenge, subcellular localization","journal":"Fish & shellfish immunology","confidence":"Medium","confidence_rationale":"Tier 1–2 — in vitro ubiquitination assay with linkage-type identification plus CRISPR KO in vivo; fish ortholog, not direct mammalian data","pmids":["40049566"],"is_preprint":false}],"current_model":"RNF182 is a brain-enriched (and more broadly expressed) RING finger E3 ubiquitin ligase that ubiquitinates multiple substrates—including ATP6V0C (targeting it for proteasomal degradation to regulate neurotransmitter release machinery), p65/NF-κB (via K48-linked ubiquitination to promote its proteasomal and/or autophagic degradation, suppressing NF-κB-driven transcription of genes such as SLC7A11 and PDL1), MFN2 (promoting mitochondrial dysfunction), and RIG-I (via K33 linkage in fish orthologs to inhibit innate antiviral signaling); its expression is transcriptionally regulated by MeCP2 and repressed by carcinogens such as BaP, and it acts upstream of the mTOR pathway in cardiac injury contexts."},"narrative":{"teleology":[{"year":2008,"claim":"Establishing RNF182 as a functional E3 ubiquitin ligase with a defined neuronal substrate resolved the question of whether this brain-enriched RING finger protein possesses catalytic activity and identified ATP6V0C as its first target for proteasomal degradation.","evidence":"In vitro ubiquitination assay, yeast two-hybrid, co-immunoprecipitation, and overexpression in cultured cells","pmids":["18298843"],"confidence":"High","gaps":["Physiological consequence of ATP6V0C degradation on neurotransmitter release not directly measured","No in vivo loss-of-function validation in neuronal tissue","Structural basis of RNF182–ATP6V0C recognition unknown"]},{"year":2010,"claim":"Identifying RNF182 as a direct transcriptional target of MeCP2 via CpG methylation connected RNF182 regulation to the epigenetic machinery governing neuronal gene expression.","evidence":"ChIP demonstrating MeCP2 binding to methylated RNF182 CpG island, bisulfite sequencing, expression profiling in MeCP2-mutant fibroblasts","pmids":["20569274"],"confidence":"Medium","gaps":["Functional consequence of MeCP2-dependent RNF182 regulation on neuronal survival not directly tested","Whether MeCP2 activates or represses RNF182 in neurons in vivo remains unclear","Single cell-line system (fibroblasts, not neurons)"]},{"year":2018,"claim":"Demonstrating that RNF182 silencing activates mTOR signaling and reduces cardiomyocyte apoptosis in ischemia-reperfusion injury placed RNF182 upstream of mTOR as a negative regulator in cardiac pathology, extending its function beyond the nervous system.","evidence":"shRNA knockdown in rat myocardial ischemia-reperfusion model with pharmacological mTOR inhibition rescue","pmids":["30450663"],"confidence":"Medium","gaps":["Direct substrate linking RNF182 to mTOR pathway suppression not identified","Single lab, no independent replication","Mechanism by which RNF182 inhibits mTOR signaling is unknown"]},{"year":2022,"claim":"Identification of p65 as a direct RNF182 substrate undergoing K48-linked ubiquitination and degradation established the molecular mechanism by which RNF182 suppresses NF-κB signaling, linking this to ferroptosis induction in hepatocellular carcinoma and to anti-inflammatory effects in tendinopathy.","evidence":"Reciprocal Co-IP, K48-linkage ubiquitination assays, ChIP on SLC7A11 promoter, functional rescue experiments in cancer cells and tenocytes, in vivo tendinopathy model","pmids":["35688944","35458235"],"confidence":"High","gaps":["Whether proteasomal vs. autophagic degradation of p65 predominates in different cell types is unresolved","Specific lysine residues on p65 targeted by RNF182 not mapped"]},{"year":2023,"claim":"Extending p65 ubiquitination to immune evasion, showing that RNF182-mediated p65 degradation suppresses PDL1 transcription and restores CD8+ T cell cytotoxicity, broadened RNF182's tumor-suppressive role to immunomodulation in lung adenocarcinoma.","evidence":"Co-IP, ChIP-qPCR on PDL1 promoter, luciferase reporter, co-culture cytotoxicity assays, in vivo xenograft","pmids":["37249301"],"confidence":"High","gaps":["Whether RNF182 affects anti-tumor immunity in immunocompetent models is untested","Contribution relative to other p65 E3 ligases not assessed"]},{"year":2023,"claim":"Demonstrating that benzo[a]pyrene represses RNF182 via AhR-dependent transcriptional repression and AhR-independent promoter hypermethylation linked carcinogen exposure to loss of RNF182 tumor-suppressive function in NSCLC.","evidence":"RT-qPCR, ChIP for AhR binding, bisulfite genomic sequencing, RNF182 overexpression rescue of proliferation in NSCLC cell lines","pmids":["36686776"],"confidence":"Medium","gaps":["In vivo relevance of BaP-mediated RNF182 silencing in lung carcinogenesis not shown","Whether RNF182 loss promotes NSCLC through p65 stabilization specifically was not tested"]},{"year":2025,"claim":"Identification of RIG-I as a substrate for K33-linked ubiquitination by the fish ortholog of RNF182 expanded its substrate repertoire to innate immune receptors and revealed a non-canonical ubiquitin linkage type for this ligase.","evidence":"Co-IP, ubiquitin linkage-type analysis, CRISPR/Cas9 knockout in rare minnow with in vivo viral challenge","pmids":["40049566"],"confidence":"Medium","gaps":["Whether mammalian RNF182 also ubiquitinates RIG-I is unknown","K33-linked ubiquitination mechanism and E2 partner not defined","Fish ortholog; direct extrapolation to human requires validation"]},{"year":2026,"claim":"Discovery that RNF182 ubiquitinates MFN2 to drive mitochondrial dysfunction in renal fibrosis identified a fourth substrate class and linked RNF182 to mitochondrial quality control.","evidence":"In vivo mouse UUO and 5/6Nx models, TGF-β1-stimulated kidney cells, RNF182 inhibition rescue, mitochondrial respiration and ATP measurements","pmids":["41461638"],"confidence":"Medium","gaps":["Ubiquitin linkage type on MFN2 not determined","Whether RNF182-MFN2 axis operates in tissues beyond kidney is unknown","Single lab finding"]},{"year":null,"claim":"The structural basis for RNF182 substrate selectivity across its diverse targets (ATP6V0C, p65, MFN2, RIG-I), its preferred E2 partners, and its physiological role in mammalian brain development remain uncharacterized.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural or biochemical reconstitution of RNF182 with any substrate","No mammalian knockout phenotype reported","E2 ubiquitin-conjugating enzyme preference not identified"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,2,3,4,6,9]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0,1,3,9]}],"localization":[],"pathway":[{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,1,2,3,4,6,9]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,2,3,5,6]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[1,5]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,9]}],"complexes":[],"partners":["RELA","ATP6V0C","MFN2","SQSTM1","MECP2"],"other_free_text":[]},"mechanistic_narrative":"RNF182 is a RING finger E3 ubiquitin ligase that ubiquitinates multiple substrates to regulate NF-κB signaling, neurotransmitter release, mitochondrial dynamics, and innate immunity. RNF182 catalyzes K48-linked polyubiquitination of the NF-κB subunit p65, targeting it for both proteasomal and p62-mediated autophagic degradation, which suppresses NF-κB-dependent transcription of genes including SLC7A11 and CD274 (PDL1) in cancer cells and inflammatory contexts [PMID:35688944, PMID:37249301, PMID:35458235, PMID:41503854]. RNF182 also ubiquitinates ATP6V0C to promote its proteasomal degradation, linking it to regulation of V-ATPase-dependent neurotransmitter release, and ubiquitinates MFN2 to drive mitochondrial dysfunction in renal fibrosis [PMID:18298843, PMID:41461638]. RNF182 expression is transcriptionally regulated by MeCP2 via CpG methylation and is repressed by the carcinogen benzo[a]pyrene through AhR-dependent and AhR-independent promoter hypermethylation, with loss of RNF182 promoting NSCLC cell proliferation [PMID:20569274, PMID:36686776]."},"prefetch_data":{"uniprot":{"accession":"Q8N6D2","full_name":"E3 ubiquitin-protein ligase RNF182","aliases":["RING finger protein 182","RING-type E3 ubiquitin transferase RNF182"],"length_aa":247,"mass_kda":27.4,"function":"E3 ubiquitin-protein ligase that mediates the ubiquitination of ATP6V0C and targets it to degradation via the ubiquitin-proteasome pathway (PubMed:18298843). Also plays a role in the inhibition of TLR-triggered innate immune response by mediating 'Lys'-48-linked ubiquitination and subsequent degradation of NF-kappa-B component RELA (PubMed:31432514)","subcellular_location":"Membrane; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q8N6D2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF182","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF182","total_profiled":1310},"omim":[{"mim_id":"621029","title":"RING FINGER PROTEIN 183; RNF183","url":"https://www.omim.org/entry/621029"},{"mim_id":"621026","title":"RING FINGER PROTEIN 182; RNF182","url":"https://www.omim.org/entry/621026"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Golgi apparatus","reliability":"Approved"},{"location":"Vesicles","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":8.0},{"tissue":"brain","ntpm":15.9},{"tissue":"retina","ntpm":8.1}],"url":"https://www.proteinatlas.org/search/RNF182"},"hgnc":{"alias_symbol":["MGC33993"],"prev_symbol":[]},"alphafold":{"accession":"Q8N6D2","domains":[{"cath_id":"3.30.40.10","chopping":"14-95","consensus_level":"high","plddt":86.7856,"start":14,"end":95},{"cath_id":"4.10.860","chopping":"176-242","consensus_level":"high","plddt":84.7707,"start":176,"end":242}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N6D2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N6D2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N6D2-F1-predicted_aligned_error_v6.png","plddt_mean":70.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF182","jax_strain_url":"https://www.jax.org/strain/search?query=RNF182"},"sequence":{"accession":"Q8N6D2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N6D2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N6D2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N6D2"}},"corpus_meta":[{"pmid":"35688944","id":"PMC_35688944","title":"PCDHB14 promotes ferroptosis and is a novel tumor suppressor in hepatocellular carcinoma.","date":"2022","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/35688944","citation_count":54,"is_preprint":false},{"pmid":"32486221","id":"PMC_32486221","title":"The Role of Tissue-Specific Ubiquitin Ligases, RNF183, RNF186, RNF182 and RNF152, in Disease and Biological Function.","date":"2020","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32486221","citation_count":49,"is_preprint":false},{"pmid":"18298843","id":"PMC_18298843","title":"A novel brain-enriched E3 ubiquitin ligase RNF182 is up regulated in the brains of Alzheimer's patients and targets ATP6V0C for degradation.","date":"2008","source":"Molecular neurodegeneration","url":"https://pubmed.ncbi.nlm.nih.gov/18298843","citation_count":49,"is_preprint":false},{"pmid":"37783761","id":"PMC_37783761","title":"Identifying potential biomarkers of idiopathic pulmonary fibrosis through machine learning analysis.","date":"2023","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/37783761","citation_count":39,"is_preprint":false},{"pmid":"20569274","id":"PMC_20569274","title":"Cell cloning-based transcriptome analysis in Rett patients: relevance to the pathogenesis of Rett syndrome of new human MeCP2 target genes.","date":"2010","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/20569274","citation_count":27,"is_preprint":false},{"pmid":"24475022","id":"PMC_24475022","title":"An integrative CGH, MSI and candidate genes methylation analysis of colorectal tumors.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/24475022","citation_count":26,"is_preprint":false},{"pmid":"35458235","id":"PMC_35458235","title":"Friedelin Alleviates the Pathogenesis of Collagenase-Induced Tendinopathy in Mice by Promoting the Selective Autophagic Degradation of p65.","date":"2022","source":"Nutrients","url":"https://pubmed.ncbi.nlm.nih.gov/35458235","citation_count":17,"is_preprint":false},{"pmid":"34652043","id":"PMC_34652043","title":"Ammonia induces autophagy via circ-IFNLR1/miR-2188-5p/RNF182 axis in tracheas of chickens.","date":"2021","source":"BioFactors (Oxford, England)","url":"https://pubmed.ncbi.nlm.nih.gov/34652043","citation_count":16,"is_preprint":false},{"pmid":"30450663","id":"PMC_30450663","title":"Activation of the mammalian target of rapamycin signaling pathway underlies a novel inhibitory role of ring finger protein 182 in ventricular remodeling after myocardial ischemia-reperfusion injury.","date":"2018","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30450663","citation_count":11,"is_preprint":false},{"pmid":"32353685","id":"PMC_32353685","title":"Identification and verification of EOMEs regulated network in Alopecia areata.","date":"2020","source":"International immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/32353685","citation_count":11,"is_preprint":false},{"pmid":"38894865","id":"PMC_38894865","title":"SIRPG promotes lung squamous cell carcinoma pathogenesis via M1 macrophages: a multi-omics study integrating data and Mendelian randomization.","date":"2024","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/38894865","citation_count":9,"is_preprint":false},{"pmid":"37249301","id":"PMC_37249301","title":"RNF182 induces p65 ubiquitination to affect PDL1 transcription and suppress immune evasion in lung adenocarcinoma.","date":"2023","source":"Immunity, inflammation and disease","url":"https://pubmed.ncbi.nlm.nih.gov/37249301","citation_count":6,"is_preprint":false},{"pmid":"36686776","id":"PMC_36686776","title":"Inhibition of RNF182 mediated by Bap promotes non-small cell lung cancer progression.","date":"2023","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/36686776","citation_count":5,"is_preprint":false},{"pmid":"36862916","id":"PMC_36862916","title":"The role of ferroptosis-related genes in airway epithelial cells of asthmatic patients based on bioinformatics.","date":"2023","source":"Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/36862916","citation_count":5,"is_preprint":false},{"pmid":"41503854","id":"PMC_41503854","title":"Friedelin Ameliorates Nucleus Pulposus Inflammation by Increasing p65 Autophagic Degradation to Inhibit NF-κB Signalling Pathway.","date":"2026","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/41503854","citation_count":1,"is_preprint":false},{"pmid":"41461638","id":"PMC_41461638","title":"IL‑1 receptor antagonism attenuates renal fibrosis via RNF182‑driven MFN2 destabilization and mitochondrial dysfunction.","date":"2025","source":"Cell death 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pneumonia.","date":"2026","source":"Microbiology spectrum","url":"https://pubmed.ncbi.nlm.nih.gov/41940665","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":12215,"output_tokens":2584,"usd":0.037702},"stage2":{"model":"claude-opus-4-6","input_tokens":5919,"output_tokens":2591,"usd":0.141555},"total_usd":0.179257,"stage1_batch_id":"msgbatch_01SUR8NDzVrNmyszWyJeCct7","stage2_batch_id":"msgbatch_01G7Yfutko3cDWzejrwH5AMx","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2008,\n      \"finding\": \"RNF182 is a brain-enriched E3 ubiquitin ligase that possesses RING finger-dependent E3 ubiquitin ligase activity, stimulating E2-dependent polyubiquitination in vitro, and directly interacts with ATP6V0C (a component of the V-ATPase involved in neurotransmitter release) to target it for proteasomal degradation.\",\n      \"method\": \"In vitro ubiquitination assay, yeast two-hybrid screening, co-precipitation (Co-IP), overexpression in cultured cells\",\n      \"journal\": \"Molecular neurodegeneration\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro enzymatic assay plus yeast two-hybrid and Co-IP orthogonal validation; foundational mechanistic paper\",\n      \"pmids\": [\"18298843\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF182 mediates K48-linked ubiquitination of p65 (NF-κB subunit), promoting its proteasomal degradation, thereby blocking p65 binding to the SLC7A11 promoter; this mechanism is activated downstream of PCDHB14 to induce ferroptosis in hepatocellular carcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, chromatin immunoprecipitation, western blot, cell viability assays\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP, ubiquitination assay, ChIP, and functional rescue experiments across multiple orthogonal methods\",\n      \"pmids\": [\"35688944\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF182 promotes ubiquitination and proteasomal degradation of p65 in lung adenocarcinoma (LUAD) cells, thereby suppressing p65-driven transcription of PDL1 and reducing immune evasion; RNF182 overexpression increased CD8+ T cell cytotoxicity against LUAD cells in co-culture.\",\n      \"method\": \"Co-immunoprecipitation, western blot, chromatin immunoprecipitation-qPCR, luciferase reporter assay, co-culture cytotoxicity assay, in vivo xenograft\",\n      \"journal\": \"Immunity, inflammation and disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (Co-IP, ChIP, luciferase, in vivo) in a single study\",\n      \"pmids\": [\"37249301\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF182 facilitates K48-linked ubiquitination of p65, promoting its autophagic (p62-mediated) degradation; the compound Friedelin recruits RNF182 to increase this ubiquitination, thereby inhibiting NF-κB signaling in tenocytes and alleviating tendinopathy.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay (K48-linkage), western blot, blockade of ubiquitination as rescue experiment, in vivo mouse tendinopathy model\",\n      \"journal\": \"Nutrients\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP and ubiquitination assay with functional rescue, single lab\",\n      \"pmids\": [\"35458235\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"RNF182 mediates ubiquitination and degradation of MFN2 (Mitofusin-2), leading to mitochondrial dysfunction; IL-1 receptor antagonist (rhIL-1Ra) suppresses TGF-β1-induced RNF182 expression, stabilizing MFN2 and reducing renal fibrosis.\",\n      \"method\": \"In vivo mouse models (UUO, 5/6Nx), in vitro TGF-β1-stimulated kidney cells, RNF182 inhibition rescue experiments, functional measurements of mitochondrial respiration and ATP production\",\n      \"journal\": \"Cell death discovery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic/pharmacological loss-of-function with defined molecular and functional phenotype, rescue experiments; single lab\",\n      \"pmids\": [\"41461638\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RNF182 silencing in a rat myocardial ischemia-reperfusion injury model activates the mTOR signaling pathway (upregulating mTOR, S6K1, eEF2, Bcl-2) and reduces cardiomyocyte apoptosis and ventricular remodeling; these protective effects are reversed by mTOR inhibition (PITE), placing RNF182 upstream of mTOR as a negative regulator.\",\n      \"method\": \"shRNA knockdown in rat MIRI model, pharmacological mTOR inhibition, cardiac function measurements, apoptosis assays, protein expression analysis\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis via pharmacological rescue in vivo, single lab, no direct substrate identification\",\n      \"pmids\": [\"30450663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"RNF182 enhances p65 association with the autophagy receptor p62 and promotes autophagic degradation of p65 in nucleus pulposus cells; Friedelin treatment increases RNF182-p65 interaction and triggers selective p65 autophagy independently of IKK activity, inhibiting NF-κB signaling in intervertebral disc degeneration.\",\n      \"method\": \"In vivo mouse cervical spine instability model, in vitro nucleus pulposus cell assays, Co-immunoprecipitation, western blot, autophagy flux assays\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — Co-IP and functional in vivo/in vitro experiments, single lab\",\n      \"pmids\": [\"41503854\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"RNF182 expression is regulated by the transcriptional modulator MeCP2; chromatin immunoprecipitation confirmed MeCP2 binding to the methylated CpG island of the RNF182 gene, identifying RNF182 as a direct MeCP2 target gene involved in neuronal cell survival.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP), bisulfite sequencing, microarray expression profiling in MeCP2 mutant vs. wild-type clonal fibroblast cultures\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — ChIP with bisulfite validation; single lab, moderate method depth\",\n      \"pmids\": [\"20569274\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Benzo[a]pyrene (BaP) suppresses RNF182 expression in NSCLC cells via two mechanisms: AhR-dependent transcriptional repression and AhR-independent promoter hypermethylation; loss of RNF182 promotes cell proliferation and cell cycle progression in NSCLC cell lines.\",\n      \"method\": \"RT-qPCR, western blot, ChIP assay, bisulfite genomic sequencing (BGS), RNF182 overexpression cell assays\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods (ChIP, BGS, functional assays) in single lab\",\n      \"pmids\": [\"36686776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Fish RNF182 (CiE3RNF182) associates with RIG-I and catalyzes Lys-33-linked polyubiquitination at the Lys33 residue of RIG-I, triggering RIG-I degradation and thereby inhibiting type I IFN antiviral signaling; CRISPR/Cas9 knockout of RNF182 in rare minnow enhanced survival against grass carp reovirus infection.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay (linkage-type analysis), CRISPR/Cas9 knockout, in vivo virus challenge, subcellular localization\",\n      \"journal\": \"Fish & shellfish immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro ubiquitination assay with linkage-type identification plus CRISPR KO in vivo; fish ortholog, not direct mammalian data\",\n      \"pmids\": [\"40049566\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF182 is a brain-enriched (and more broadly expressed) RING finger E3 ubiquitin ligase that ubiquitinates multiple substrates—including ATP6V0C (targeting it for proteasomal degradation to regulate neurotransmitter release machinery), p65/NF-κB (via K48-linked ubiquitination to promote its proteasomal and/or autophagic degradation, suppressing NF-κB-driven transcription of genes such as SLC7A11 and PDL1), MFN2 (promoting mitochondrial dysfunction), and RIG-I (via K33 linkage in fish orthologs to inhibit innate antiviral signaling); its expression is transcriptionally regulated by MeCP2 and repressed by carcinogens such as BaP, and it acts upstream of the mTOR pathway in cardiac injury contexts.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RNF182 is a RING finger E3 ubiquitin ligase that ubiquitinates multiple substrates to regulate NF-κB signaling, neurotransmitter release, mitochondrial dynamics, and innate immunity. RNF182 catalyzes K48-linked polyubiquitination of the NF-κB subunit p65, targeting it for both proteasomal and p62-mediated autophagic degradation, which suppresses NF-κB-dependent transcription of genes including SLC7A11 and CD274 (PDL1) in cancer cells and inflammatory contexts [PMID:35688944, PMID:37249301, PMID:35458235, PMID:41503854]. RNF182 also ubiquitinates ATP6V0C to promote its proteasomal degradation, linking it to regulation of V-ATPase-dependent neurotransmitter release, and ubiquitinates MFN2 to drive mitochondrial dysfunction in renal fibrosis [PMID:18298843, PMID:41461638]. RNF182 expression is transcriptionally regulated by MeCP2 via CpG methylation and is repressed by the carcinogen benzo[a]pyrene through AhR-dependent and AhR-independent promoter hypermethylation, with loss of RNF182 promoting NSCLC cell proliferation [PMID:20569274, PMID:36686776].\",\n  \"teleology\": [\n    {\n      \"year\": 2008,\n      \"claim\": \"Establishing RNF182 as a functional E3 ubiquitin ligase with a defined neuronal substrate resolved the question of whether this brain-enriched RING finger protein possesses catalytic activity and identified ATP6V0C as its first target for proteasomal degradation.\",\n      \"evidence\": \"In vitro ubiquitination assay, yeast two-hybrid, co-immunoprecipitation, and overexpression in cultured cells\",\n      \"pmids\": [\"18298843\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Physiological consequence of ATP6V0C degradation on neurotransmitter release not directly measured\",\n        \"No in vivo loss-of-function validation in neuronal tissue\",\n        \"Structural basis of RNF182–ATP6V0C recognition unknown\"\n      ]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identifying RNF182 as a direct transcriptional target of MeCP2 via CpG methylation connected RNF182 regulation to the epigenetic machinery governing neuronal gene expression.\",\n      \"evidence\": \"ChIP demonstrating MeCP2 binding to methylated RNF182 CpG island, bisulfite sequencing, expression profiling in MeCP2-mutant fibroblasts\",\n      \"pmids\": [\"20569274\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Functional consequence of MeCP2-dependent RNF182 regulation on neuronal survival not directly tested\",\n        \"Whether MeCP2 activates or represses RNF182 in neurons in vivo remains unclear\",\n        \"Single cell-line system (fibroblasts, not neurons)\"\n      ]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Demonstrating that RNF182 silencing activates mTOR signaling and reduces cardiomyocyte apoptosis in ischemia-reperfusion injury placed RNF182 upstream of mTOR as a negative regulator in cardiac pathology, extending its function beyond the nervous system.\",\n      \"evidence\": \"shRNA knockdown in rat myocardial ischemia-reperfusion model with pharmacological mTOR inhibition rescue\",\n      \"pmids\": [\"30450663\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Direct substrate linking RNF182 to mTOR pathway suppression not identified\",\n        \"Single lab, no independent replication\",\n        \"Mechanism by which RNF182 inhibits mTOR signaling is unknown\"\n      ]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identification of p65 as a direct RNF182 substrate undergoing K48-linked ubiquitination and degradation established the molecular mechanism by which RNF182 suppresses NF-κB signaling, linking this to ferroptosis induction in hepatocellular carcinoma and to anti-inflammatory effects in tendinopathy.\",\n      \"evidence\": \"Reciprocal Co-IP, K48-linkage ubiquitination assays, ChIP on SLC7A11 promoter, functional rescue experiments in cancer cells and tenocytes, in vivo tendinopathy model\",\n      \"pmids\": [\"35688944\", \"35458235\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether proteasomal vs. autophagic degradation of p65 predominates in different cell types is unresolved\",\n        \"Specific lysine residues on p65 targeted by RNF182 not mapped\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extending p65 ubiquitination to immune evasion, showing that RNF182-mediated p65 degradation suppresses PDL1 transcription and restores CD8+ T cell cytotoxicity, broadened RNF182's tumor-suppressive role to immunomodulation in lung adenocarcinoma.\",\n      \"evidence\": \"Co-IP, ChIP-qPCR on PDL1 promoter, luciferase reporter, co-culture cytotoxicity assays, in vivo xenograft\",\n      \"pmids\": [\"37249301\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Whether RNF182 affects anti-tumor immunity in immunocompetent models is untested\",\n        \"Contribution relative to other p65 E3 ligases not assessed\"\n      ]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Demonstrating that benzo[a]pyrene represses RNF182 via AhR-dependent transcriptional repression and AhR-independent promoter hypermethylation linked carcinogen exposure to loss of RNF182 tumor-suppressive function in NSCLC.\",\n      \"evidence\": \"RT-qPCR, ChIP for AhR binding, bisulfite genomic sequencing, RNF182 overexpression rescue of proliferation in NSCLC cell lines\",\n      \"pmids\": [\"36686776\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"In vivo relevance of BaP-mediated RNF182 silencing in lung carcinogenesis not shown\",\n        \"Whether RNF182 loss promotes NSCLC through p65 stabilization specifically was not tested\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identification of RIG-I as a substrate for K33-linked ubiquitination by the fish ortholog of RNF182 expanded its substrate repertoire to innate immune receptors and revealed a non-canonical ubiquitin linkage type for this ligase.\",\n      \"evidence\": \"Co-IP, ubiquitin linkage-type analysis, CRISPR/Cas9 knockout in rare minnow with in vivo viral challenge\",\n      \"pmids\": [\"40049566\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Whether mammalian RNF182 also ubiquitinates RIG-I is unknown\",\n        \"K33-linked ubiquitination mechanism and E2 partner not defined\",\n        \"Fish ortholog; direct extrapolation to human requires validation\"\n      ]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Discovery that RNF182 ubiquitinates MFN2 to drive mitochondrial dysfunction in renal fibrosis identified a fourth substrate class and linked RNF182 to mitochondrial quality control.\",\n      \"evidence\": \"In vivo mouse UUO and 5/6Nx models, TGF-β1-stimulated kidney cells, RNF182 inhibition rescue, mitochondrial respiration and ATP measurements\",\n      \"pmids\": [\"41461638\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Ubiquitin linkage type on MFN2 not determined\",\n        \"Whether RNF182-MFN2 axis operates in tissues beyond kidney is unknown\",\n        \"Single lab finding\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The structural basis for RNF182 substrate selectivity across its diverse targets (ATP6V0C, p65, MFN2, RIG-I), its preferred E2 partners, and its physiological role in mammalian brain development remain uncharacterized.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No structural or biochemical reconstitution of RNF182 with any substrate\",\n        \"No mammalian knockout phenotype reported\",\n        \"E2 ubiquitin-conjugating enzyme preference not identified\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 6, 9]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 1, 3, 9]}\n    ],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 1, 2, 3, 4, 6, 9]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 2, 3, 5, 6]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [1, 5]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 9]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"RELA\",\n      \"ATP6V0C\",\n      \"MFN2\",\n      \"SQSTM1\",\n      \"MECP2\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}