{"gene":"RNF183","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":2016,"finding":"RNF183 promotes intestinal inflammation by ubiquitinating IκBα, leading to its degradation and consequent NF-κB activation; this activity is negatively regulated by miR-7, which suppresses RNF183 expression","method":"siRNA knockdown, plasmid transfection, immunoprecipitation, miRNA mimic/inhibitor transfection","journal":"Journal of Crohn's & colitis","confidence":"Medium","confidence_rationale":"Tier 2 — reciprocal Co-IP and functional knockdown with defined phenotype, single lab","pmids":["26818663"],"is_preprint":false},{"year":2017,"finding":"RNF183 activates the NF-κB signaling pathway through p65 and stimulates IL-8 transcription to promote colorectal cancer cell migration and invasion; this function requires its E3 ubiquitin ligase activity","method":"siRNA knockdown, small molecule NF-κB inhibitor, E3 ligase activity-dead mutant, in vitro migration/invasion assays, in vivo xenograft","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function with defined cellular phenotype and E3 ligase mutant validation, single lab","pmids":["28796265"],"is_preprint":false},{"year":2018,"finding":"RNF183 is an ER-localized transmembrane RING finger E3 ubiquitin ligase that is upregulated by sustained ER stress in an IRE1α-dependent manner (via IRE1-mediated reduction of miR-7); RNF183 interacts with and polyubiquitinates Bcl-xL, targeting it for proteasomal degradation and thereby inducing apoptosis","method":"Co-immunoprecipitation, in vitro ubiquitination assay, subcellular fractionation/fluorescence imaging, siRNA knockdown, overexpression, IRE1α inhibition","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1-2 — in vitro E3 ligase activity, Co-IP of substrate, mechanistic dissection of upstream regulation, multiple orthogonal methods","pmids":["29507230"],"is_preprint":false},{"year":2018,"finding":"RNF183 predominantly localizes to the ER, Golgi, and lysosome; it interacts with Sec16A (a COPII vesicle formation protein) via Sec16A's central conserved domain, and Sec16A stabilizes RNF183 by protecting it from ERAD-mediated degradation","method":"Co-immunoprecipitation, subcellular localization imaging, siRNA knockdown, ERAD assay","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2-3 — Co-IP and localization experiments with functional stability consequence, single lab","pmids":["29300766"],"is_preprint":false},{"year":2018,"finding":"NFAT5 transcription factor directly binds to a conserved motif in the RNF183 promoter (-3,466 to -3,136 bp upstream) and drives kidney-specific, hypertonicity-induced RNF183 expression; RNF183 knockdown increases hypertonicity-induced apoptosis in renal medullary cells","method":"siRNA knockdown, luciferase reporter assay, ChIP assay, promoter mutation analysis, caspase-3 activation assay","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1-2 — ChIP, reporter assay with binding site mutation, and functional consequence, multiple orthogonal methods in single lab","pmids":["30413537"],"is_preprint":false},{"year":2019,"finding":"RNF183 mediates K63-linked ubiquitination of death receptor 5 (DR5), targeting it for lysosomal degradation; this promotes TRAIL-induced caspase-8 and caspase-3 activation and apoptosis","method":"Co-immunoprecipitation, ubiquitination assay (K63-linkage specific), lysosomal inhibitor treatment, caspase activation assay, siRNA knockdown","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1-2 — linkage-specific ubiquitination assay, Co-IP of substrate, functional caspase readout, multiple orthogonal methods","pmids":["31889078"],"is_preprint":false},{"year":2019,"finding":"RNF183 ubiquitinates the Na,K-ATPase β1 subunit (but not α1), causing lysosomal co-degradation of both α1 and β1 subunits; RNF183 translocates Na,K-ATPase from the plasma membrane to lysosomes under hypertonic conditions","method":"BirA proximity biotinylation, Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, chloroquine lysosomal inhibitor treatment, stable cell line overexpression","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1-2 — proximity biotinylation for substrate ID, Co-IP, ubiquitination assay, lysosomal degradation confirmation, multiple methods","pmids":["31732153"],"is_preprint":false},{"year":2019,"finding":"RNF183 expression in renal medullary collecting ducts is regulated in vivo by NFAT5 in response to tonicity; furosemide treatment decreases renal medullary tonicity, reduces NFAT5 activity, and decreases RNF183 expression; RNF183-GFP knock-in mice show RNF183 colocalizes with aquaporin-2 in collecting duct cells","method":"CRISPR/Cas9 knock-in mice (RNF183-GFP), immunofluorescence, furosemide in vivo treatment, mIMCD-3 cell reporter assay","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1-2 — in vivo knock-in mouse model with furosemide perturbation, direct colocalization with collecting duct marker","pmids":["31053298"],"is_preprint":false},{"year":2022,"finding":"RNF183 ubiquitinates SHP2 (a negative regulator of STAT3), leading to its degradation and consequent STAT3 pathway activation, which promotes lung adenocarcinoma cell proliferation","method":"Co-immunoprecipitation, ubiquitination assay, siRNA knockdown, in vivo xenograft model","journal":"Cell cycle (Georgetown, Tex.)","confidence":"Medium","confidence_rationale":"Tier 2-3 — Co-IP and ubiquitination assay with functional downstream readout, single lab","pmids":["35104174"],"is_preprint":false},{"year":2022,"finding":"Cathepsin D inhibition upregulates RNF183 expression via NF-κB activation, which in turn destabilizes Bcl-xL through RNF183 E3 ligase activity, sensitizing cancer cells to anticancer drug-induced apoptosis","method":"siRNA knockdown, western blot, xenograft model, correlation analysis in tissue samples","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2-3 — pathway placement via genetic knockdown with apoptosis readout, supported by in vivo data, single lab","pmids":["35121737"],"is_preprint":false},{"year":2024,"finding":"FBXO5, an F-box protein whose expression is regulated by E2F2 downstream of ER stress, directly binds RNF183 and promotes its ubiquitin-dependent proteasomal degradation, thereby counteracting RNF183-mediated apoptosis; FBXO5 ΔF-box mutant cannot degrade RNF183, confirming SCF complex requirement","method":"Co-immunoprecipitation, ubiquitination assay, F-box deletion mutant, siRNA knockdown, overexpression rescue experiments, in vivo tumor model","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 1-2 — direct binding shown by Co-IP, F-box mutant validation, rescue experiment confirming epistasis, multiple orthogonal methods","pmids":["38212299"],"is_preprint":false},{"year":2025,"finding":"RNF183 directly ubiquitinates PKM2 (M2 pyruvate kinase), targeting it for proteasomal degradation; RNF183 expression is regulated by m6A methylation via METTL3 writer and IGF2BP2 reader; in podocytes under high glucose, reduced METTL3/IGF2BP2 lowers RNF183, decreasing PKM2 ubiquitination and aggravating podocyte injury","method":"Co-immunoprecipitation, LC-MS, ubiquitination assay, m6A inhibitor (MA2) treatment, overexpression/knockdown, transcriptome sequencing","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 1-2 — Co-IP plus LC-MS for substrate identification and ubiquitination confirmation, single lab","pmids":["40072093"],"is_preprint":false},{"year":2026,"finding":"RNF183 promotes K63-linked ubiquitination of NKCC1 (Na-K-2Cl co-transporter 1), leading to its lysosomal degradation; this regulates intracellular Na+ homeostasis under hypertonic conditions in renal collecting duct cells, and knockout of RNF183 in mIMCD-3 cells increases hypertonic-induced caspase-3 cleavage and cell death","method":"Co-immunoprecipitation, K63-linkage specific ubiquitination assay, lysosomal inhibitor treatment, CRISPR/Cas9 knockout, intracellular Na+ measurement, caspase-3 cleavage assay","journal":"Communications biology","confidence":"High","confidence_rationale":"Tier 1-2 — linkage-specific ubiquitination assay, CRISPR KO with defined functional phenotype, multiple orthogonal methods","pmids":["41942761"],"is_preprint":false}],"current_model":"RNF183 is a transmembrane RING finger E3 ubiquitin ligase that localizes to the ER, Golgi, and lysosomes; it ubiquitinates multiple substrates—including IκBα (K48-linked, proteasomal), Bcl-xL (proteasomal), DR5 (K63-linked, lysosomal), Na,K-ATPase β1 (lysosomal), NKCC1 (K63-linked, lysosomal), SHP2 (proteasomal), and PKM2 (proteasomal)—thereby regulating NF-κB-mediated inflammation, ER stress-induced apoptosis, TRAIL/caspase signaling, ion transporter turnover, and STAT3 pathway activity; its expression in the renal medulla is transcriptionally driven by NFAT5 in response to hypertonicity, post-transcriptionally stabilized via IRE1α-mediated suppression of miR-7, and its protein stability is controlled by Sec16A (protection from ERAD) and FBXO5-SCF complex (degradation)."},"narrative":{"teleology":[{"year":2016,"claim":"Establishing that RNF183 functions as a pro-inflammatory E3 ligase by identifying IκBα as a direct ubiquitination substrate whose degradation activates NF-κB, and that miR-7 negatively regulates RNF183 expression, provided the first mechanistic link between this RING finger protein and intestinal inflammation.","evidence":"siRNA knockdown, overexpression, Co-IP of IκBα, miR-7 mimic/inhibitor experiments in intestinal cell lines","pmids":["26818663"],"confidence":"Medium","gaps":["IκBα ubiquitination linkage type not specified","no in vivo intestinal inflammation model","single-lab finding without independent replication"]},{"year":2017,"claim":"Demonstrating that the NF-κB–activating function of RNF183 requires intact E3 ligase catalytic activity and drives IL-8 transcription to promote colorectal cancer cell migration and invasion extended the pro-inflammatory role to a cancer-promoting context.","evidence":"E3 ligase activity-dead mutant, NF-κB inhibitor, migration/invasion assays, xenograft model in colorectal cancer cells","pmids":["28796265"],"confidence":"Medium","gaps":["direct ubiquitination substrate in this cancer context not formally identified","single-lab study"]},{"year":2018,"claim":"Identification of RNF183 as an ER-localized transmembrane E3 ligase upregulated by sustained ER stress through IRE1α-mediated suppression of miR-7, with Bcl-xL as a direct ubiquitination substrate targeted for proteasomal degradation, established the mechanistic basis for ER stress–induced apoptosis via RNF183.","evidence":"In vitro ubiquitination assay, Co-IP, subcellular fractionation, IRE1α inhibition, fluorescence imaging in multiple cell lines","pmids":["29507230"],"confidence":"High","gaps":["ubiquitination site on Bcl-xL not mapped","contribution relative to other Bcl-xL regulatory mechanisms not quantified"]},{"year":2018,"claim":"Discovery that Sec16A physically interacts with RNF183 and protects it from ERAD-mediated degradation, together with confirmation that RNF183 localizes to the ER, Golgi, and lysosomes, revealed a protein stability control mechanism and clarified the subcellular distribution enabling RNF183's diverse substrate access.","evidence":"Co-IP, subcellular localization imaging, siRNA knockdown of Sec16A with ERAD readout","pmids":["29300766"],"confidence":"Medium","gaps":["structural basis of Sec16A–RNF183 interaction not defined","single-lab study without independent confirmation"]},{"year":2018,"claim":"Showing that NFAT5 directly binds the RNF183 promoter to drive kidney-specific, hypertonicity-induced expression established the transcriptional logic governing RNF183 in the renal medulla and linked its loss to increased apoptosis under osmotic stress.","evidence":"ChIP, luciferase reporter with binding-site mutation, caspase-3 activation in mIMCD-3 cells","pmids":["30413537"],"confidence":"High","gaps":["identity of the RNF183 substrate(s) mediating renal anti-apoptotic protection was unknown at this stage"]},{"year":2019,"claim":"Identification of DR5 and Na,K-ATPase β1 as K63-linked and lysosomal-targeted ubiquitination substrates expanded RNF183's repertoire beyond proteasomal degradation and showed it regulates both TRAIL-mediated apoptosis and plasma membrane ion transporter turnover under hypertonic conditions.","evidence":"K63-linkage-specific ubiquitination assays, lysosomal inhibitor rescue, BirA proximity biotinylation (Na,K-ATPase β1), Co-IP, caspase activation assays","pmids":["31889078","31732153"],"confidence":"High","gaps":["how RNF183 distinguishes between K48- and K63-linkage on different substrates is unknown","E2 conjugating enzyme identity not determined"]},{"year":2019,"claim":"In vivo validation using CRISPR/Cas9 RNF183-GFP knock-in mice confirmed renal medullary collecting duct expression, NFAT5-dependent regulation by tonicity (furosemide treatment), and colocalization with aquaporin-2, establishing the physiological context of RNF183 function in the kidney.","evidence":"CRISPR/Cas9 knock-in mouse, immunofluorescence, furosemide in vivo perturbation","pmids":["31053298"],"confidence":"High","gaps":["no full knockout mouse phenotype reported","functional consequence of RNF183 loss in vivo not assessed at this stage"]},{"year":2022,"claim":"Demonstrating that RNF183 ubiquitinates and degrades SHP2 to activate STAT3 signaling and promote lung adenocarcinoma proliferation broadened the oncogenic signaling roles of RNF183 beyond NF-κB.","evidence":"Co-IP, ubiquitination assay, siRNA knockdown, in vivo xenograft in lung adenocarcinoma cells","pmids":["35104174"],"confidence":"Medium","gaps":["ubiquitin linkage type on SHP2 not determined","single-lab finding"]},{"year":2022,"claim":"Placing RNF183 downstream of cathepsin D inhibition in a positive feedback loop with NF-κB, where NF-κB induces RNF183 which then degrades Bcl-xL, clarified how RNF183 can be pharmacologically leveraged to sensitize cancer cells to apoptosis.","evidence":"siRNA knockdown, western blot, xenograft model, tissue correlation analysis","pmids":["35121737"],"confidence":"Medium","gaps":["whether NF-κB directly binds the RNF183 promoter was not demonstrated","single-lab study"]},{"year":2024,"claim":"Identifying FBXO5–SCF as the E3 ligase complex that ubiquitinates and degrades RNF183 itself, regulated by E2F2 during ER stress, established a negative feedback circuit controlling RNF183 protein levels and apoptotic output.","evidence":"Co-IP, F-box deletion mutant, ubiquitination assay, rescue experiments, in vivo tumor model","pmids":["38212299"],"confidence":"High","gaps":["ubiquitination sites on RNF183 targeted by FBXO5 not mapped","interplay between FBXO5-mediated and Sec16A-mediated stability control not addressed"]},{"year":2025,"claim":"Discovery that RNF183 directly ubiquitinates PKM2 for proteasomal degradation and that RNF183 mRNA stability is regulated by METTL3/IGF2BP2-dependent m6A modification in podocytes added an epitranscriptomic regulatory layer and a metabolic substrate to RNF183 biology.","evidence":"Co-IP, LC-MS substrate identification, ubiquitination assay, m6A inhibitor treatment, overexpression/knockdown in podocytes under high glucose","pmids":["40072093"],"confidence":"Medium","gaps":["m6A modification sites on RNF183 mRNA not mapped","in vivo diabetic nephropathy model not included","single-lab finding"]},{"year":2026,"claim":"Identification of NKCC1 as a K63-linked ubiquitination substrate directed to lysosomal degradation by RNF183, combined with CRISPR knockout evidence that RNF183 loss increases hypertonic cell death, established RNF183 as a key regulator of intracellular Na⁺ homeostasis and cell survival in renal collecting duct cells.","evidence":"K63-linkage-specific ubiquitination assay, CRISPR/Cas9 knockout in mIMCD-3 cells, lysosomal inhibitor rescue, intracellular Na⁺ measurement, caspase-3 cleavage assay","pmids":["41942761"],"confidence":"High","gaps":["in vivo renal phenotype of RNF183 knockout not yet reported","how RNF183 is itself regulated under hypertonicity at the protein level (beyond NFAT5 transcription) remains unclear"]},{"year":null,"claim":"The E2 conjugating enzyme(s) used by RNF183, the structural basis for its dual K48/K63 linkage specificity, and the full phenotype of RNF183 loss in a whole-organism model remain unresolved.","evidence":"","pmids":[],"confidence":"High","gaps":["E2 enzyme partner(s) not identified","no crystal or cryo-EM structure of RNF183 or RNF183–substrate complexes","no complete knockout mouse phenotypic characterization","how substrate selection between K48 and K63 linkage is determined is unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,1,2,5,6,8,11,12]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0,2,5,6,8,11,12]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[2,3]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[3]},{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[2,5,9]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,2,5,6,8,10,11,12]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[0,1,8]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[0,1]},{"term_id":"R-HSA-8953897","term_label":"Cellular responses to stimuli","supporting_discovery_ids":[2,4,9]},{"term_id":"R-HSA-382551","term_label":"Transport of small molecules","supporting_discovery_ids":[6,12]}],"complexes":[],"partners":["NFKBIA","BCL2L1","TNFRSF10B","ATP1B1","SLC12A2","PTPN11","PKM","SEC16A"],"other_free_text":[]},"mechanistic_narrative":"RNF183 is a transmembrane RING finger E3 ubiquitin ligase that controls cell survival, inflammatory signaling, and ion transporter turnover by targeting multiple substrates for either proteasomal or lysosomal degradation. It catalyzes K48-linked polyubiquitination of IκBα, Bcl-xL, SHP2, and PKM2 to promote their proteasomal destruction—thereby activating NF-κB signaling, sensitizing cells to ER stress–induced apoptosis, enhancing STAT3 pathway activity, and modulating glycolytic metabolism, respectively [PMID:26818663, PMID:29507230, PMID:35104174, PMID:40072093]—and K63-linked polyubiquitination of DR5, Na,K-ATPase β1, and NKCC1 to direct their lysosomal degradation, regulating TRAIL-induced apoptosis and renal ion homeostasis under hypertonic stress [PMID:31889078, PMID:31732153, PMID:41942761]. RNF183 expression is transcriptionally driven by NFAT5 in the renal medulla in response to hypertonicity, post-transcriptionally stabilized by IRE1α-mediated suppression of miR-7 during ER stress, epitranscriptomically regulated by METTL3/IGF2BP2-dependent m6A modification, and its protein stability is governed by Sec16A-mediated protection from ERAD and FBXO5–SCF complex–mediated proteasomal degradation [PMID:30413537, PMID:29507230, PMID:40072093, PMID:29300766, PMID:38212299]."},"prefetch_data":{"uniprot":{"accession":"Q96D59","full_name":"E3 ubiquitin-protein ligase RNF183","aliases":[],"length_aa":192,"mass_kda":21.6,"function":"Acts as an E3 ubiquitin ligase catalyzing the covalent attachment of ubiquitin moieties onto substrate proteins (PubMed:29507230). Triggers apoptosis in response to prolonged ER stress by mediating the polyubiquitination and subsequent proteasomal degradation of BCL2L1 (PubMed:29507230). May collaborate with FATE1 to restrain BIK protein levels thus regulating apoptotic signaling (PubMed:26567849)","subcellular_location":"Endoplasmic reticulum membrane; Endoplasmic reticulum; Golgi apparatus, cis-Golgi network membrane; Lysosome membrane","url":"https://www.uniprot.org/uniprotkb/Q96D59/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF183","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/RNF183","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":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in some","driving_tissues":[{"tissue":"epididymis","ntpm":15.3},{"tissue":"kidney","ntpm":9.3}],"url":"https://www.proteinatlas.org/search/RNF183"},"hgnc":{"alias_symbol":["MGC4734"],"prev_symbol":[]},"alphafold":{"accession":"Q96D59","domains":[{"cath_id":"3.30.40.10","chopping":"9-89","consensus_level":"medium","plddt":82.0569,"start":9,"end":89},{"cath_id":"1.20.5","chopping":"157-192","consensus_level":"medium","plddt":80.8286,"start":157,"end":192}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96D59","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q96D59-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q96D59-F1-predicted_aligned_error_v6.png","plddt_mean":67.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF183","jax_strain_url":"https://www.jax.org/strain/search?query=RNF183"},"sequence":{"accession":"Q96D59","fasta_url":"https://rest.uniprot.org/uniprotkb/Q96D59.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q96D59/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q96D59"}},"corpus_meta":[{"pmid":"28796265","id":"PMC_28796265","title":"RNF183 promotes proliferation and metastasis of colorectal cancer cells via activation of NF-κB-IL-8 axis.","date":"2017","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/28796265","citation_count":58,"is_preprint":false},{"pmid":"26818663","id":"PMC_26818663","title":"E3 Ubiquitin ligase RNF183 Is a Novel Regulator in Inflammatory Bowel Disease.","date":"2016","source":"Journal of Crohn's & colitis","url":"https://pubmed.ncbi.nlm.nih.gov/26818663","citation_count":55,"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":"29507230","id":"PMC_29507230","title":"Transmembrane E3 ligase RNF183 mediates ER stress-induced apoptosis by degrading Bcl-xL.","date":"2018","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/29507230","citation_count":44,"is_preprint":false},{"pmid":"35121737","id":"PMC_35121737","title":"Cathepsin D as a potential therapeutic target to enhance anticancer drug-induced apoptosis via RNF183-mediated destabilization of Bcl-xL in cancer cells.","date":"2022","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/35121737","citation_count":36,"is_preprint":false},{"pmid":"37258572","id":"PMC_37258572","title":"The YTHDC1/GLUT3/RNF183 axis forms a positive feedback loop that modulates glucose metabolism and bladder cancer progression.","date":"2023","source":"Experimental & molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/37258572","citation_count":29,"is_preprint":false},{"pmid":"38212299","id":"PMC_38212299","title":"FBXO5-mediated RNF183 degradation prevents endoplasmic reticulum stress-induced apoptosis and promotes colon cancer progression.","date":"2024","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/38212299","citation_count":18,"is_preprint":false},{"pmid":"30413537","id":"PMC_30413537","title":"NFAT5 up-regulates expression of the kidney-specific ubiquitin ligase gene Rnf183 under hypertonic conditions in inner-medullary collecting duct cells.","date":"2018","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/30413537","citation_count":18,"is_preprint":false},{"pmid":"31889078","id":"PMC_31889078","title":"Inflammatory bowel disease-associated ubiquitin ligase RNF183 promotes lysosomal degradation of DR5 and TRAIL-induced caspase activation.","date":"2019","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/31889078","citation_count":14,"is_preprint":false},{"pmid":"29300766","id":"PMC_29300766","title":"Sec16A, a key protein in COPII vesicle formation, regulates the stability and localization of the novel ubiquitin ligase RNF183.","date":"2018","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/29300766","citation_count":11,"is_preprint":false},{"pmid":"34777536","id":"PMC_34777536","title":"Moxibustion Inhibits the Expression of Colonic NLRP3 through miR7/RNF183/NF-κB Signaling Pathway in UC Rats.","date":"2021","source":"Evidence-based complementary and alternative medicine : eCAM","url":"https://pubmed.ncbi.nlm.nih.gov/34777536","citation_count":10,"is_preprint":false},{"pmid":"31732153","id":"PMC_31732153","title":"Hypertonicity-responsive ubiquitin ligase RNF183 promotes Na, K-ATPase lysosomal degradation through ubiquitination of its β1 subunit.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31732153","citation_count":10,"is_preprint":false},{"pmid":"31053298","id":"PMC_31053298","title":"Renal medullary tonicity regulates RNF183 expression in the collecting ducts via NFAT5.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31053298","citation_count":7,"is_preprint":false},{"pmid":"35104174","id":"PMC_35104174","title":"Knockdown of RNF183 suppressed proliferation of lung adenocarcinoma cells via inactivating the STAT3 signaling pathway.","date":"2022","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/35104174","citation_count":5,"is_preprint":false},{"pmid":"38044546","id":"PMC_38044546","title":"RNF183 Promotes Colon Cancer Cell Stemness through Fatty Acid Oxidation.","date":"2024","source":"Nutrition and cancer","url":"https://pubmed.ncbi.nlm.nih.gov/38044546","citation_count":4,"is_preprint":false},{"pmid":"40072093","id":"PMC_40072093","title":"Modification of RNF183 via m6A Methylation Mediates Podocyte Dysfunction in Diabetic Nephropathy by Regulating PKM2 Ubiquitination and Degradation.","date":"2025","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/40072093","citation_count":3,"is_preprint":false},{"pmid":"41942761","id":"PMC_41942761","title":"Hypertonicity-induced RNF183 targets NKCC1 to protect kidney collecting duct cells but induces apoptosis in colon cells.","date":"2026","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/41942761","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":10412,"output_tokens":3263,"usd":0.040091},"stage2":{"model":"claude-opus-4-6","input_tokens":6688,"output_tokens":6415,"usd":0.290722},"total_usd":0.330813,"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\": 2016,\n      \"finding\": \"RNF183 promotes intestinal inflammation by ubiquitinating IκBα, leading to its degradation and consequent NF-κB activation; this activity is negatively regulated by miR-7, which suppresses RNF183 expression\",\n      \"method\": \"siRNA knockdown, plasmid transfection, immunoprecipitation, miRNA mimic/inhibitor transfection\",\n      \"journal\": \"Journal of Crohn's & colitis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal Co-IP and functional knockdown with defined phenotype, single lab\",\n      \"pmids\": [\"26818663\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RNF183 activates the NF-κB signaling pathway through p65 and stimulates IL-8 transcription to promote colorectal cancer cell migration and invasion; this function requires its E3 ubiquitin ligase activity\",\n      \"method\": \"siRNA knockdown, small molecule NF-κB inhibitor, E3 ligase activity-dead mutant, in vitro migration/invasion assays, in vivo xenograft\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function with defined cellular phenotype and E3 ligase mutant validation, single lab\",\n      \"pmids\": [\"28796265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RNF183 is an ER-localized transmembrane RING finger E3 ubiquitin ligase that is upregulated by sustained ER stress in an IRE1α-dependent manner (via IRE1-mediated reduction of miR-7); RNF183 interacts with and polyubiquitinates Bcl-xL, targeting it for proteasomal degradation and thereby inducing apoptosis\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay, subcellular fractionation/fluorescence imaging, siRNA knockdown, overexpression, IRE1α inhibition\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vitro E3 ligase activity, Co-IP of substrate, mechanistic dissection of upstream regulation, multiple orthogonal methods\",\n      \"pmids\": [\"29507230\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RNF183 predominantly localizes to the ER, Golgi, and lysosome; it interacts with Sec16A (a COPII vesicle formation protein) via Sec16A's central conserved domain, and Sec16A stabilizes RNF183 by protecting it from ERAD-mediated degradation\",\n      \"method\": \"Co-immunoprecipitation, subcellular localization imaging, siRNA knockdown, ERAD assay\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — Co-IP and localization experiments with functional stability consequence, single lab\",\n      \"pmids\": [\"29300766\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NFAT5 transcription factor directly binds to a conserved motif in the RNF183 promoter (-3,466 to -3,136 bp upstream) and drives kidney-specific, hypertonicity-induced RNF183 expression; RNF183 knockdown increases hypertonicity-induced apoptosis in renal medullary cells\",\n      \"method\": \"siRNA knockdown, luciferase reporter assay, ChIP assay, promoter mutation analysis, caspase-3 activation assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — ChIP, reporter assay with binding site mutation, and functional consequence, multiple orthogonal methods in single lab\",\n      \"pmids\": [\"30413537\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RNF183 mediates K63-linked ubiquitination of death receptor 5 (DR5), targeting it for lysosomal degradation; this promotes TRAIL-induced caspase-8 and caspase-3 activation and apoptosis\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay (K63-linkage specific), lysosomal inhibitor treatment, caspase activation assay, siRNA knockdown\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — linkage-specific ubiquitination assay, Co-IP of substrate, functional caspase readout, multiple orthogonal methods\",\n      \"pmids\": [\"31889078\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RNF183 ubiquitinates the Na,K-ATPase β1 subunit (but not α1), causing lysosomal co-degradation of both α1 and β1 subunits; RNF183 translocates Na,K-ATPase from the plasma membrane to lysosomes under hypertonic conditions\",\n      \"method\": \"BirA proximity biotinylation, Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, chloroquine lysosomal inhibitor treatment, stable cell line overexpression\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — proximity biotinylation for substrate ID, Co-IP, ubiquitination assay, lysosomal degradation confirmation, multiple methods\",\n      \"pmids\": [\"31732153\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RNF183 expression in renal medullary collecting ducts is regulated in vivo by NFAT5 in response to tonicity; furosemide treatment decreases renal medullary tonicity, reduces NFAT5 activity, and decreases RNF183 expression; RNF183-GFP knock-in mice show RNF183 colocalizes with aquaporin-2 in collecting duct cells\",\n      \"method\": \"CRISPR/Cas9 knock-in mice (RNF183-GFP), immunofluorescence, furosemide in vivo treatment, mIMCD-3 cell reporter assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — in vivo knock-in mouse model with furosemide perturbation, direct colocalization with collecting duct marker\",\n      \"pmids\": [\"31053298\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF183 ubiquitinates SHP2 (a negative regulator of STAT3), leading to its degradation and consequent STAT3 pathway activation, which promotes lung adenocarcinoma cell proliferation\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, siRNA knockdown, in vivo xenograft model\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — Co-IP and ubiquitination assay with functional downstream readout, single lab\",\n      \"pmids\": [\"35104174\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Cathepsin D inhibition upregulates RNF183 expression via NF-κB activation, which in turn destabilizes Bcl-xL through RNF183 E3 ligase activity, sensitizing cancer cells to anticancer drug-induced apoptosis\",\n      \"method\": \"siRNA knockdown, western blot, xenograft model, correlation analysis in tissue samples\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — pathway placement via genetic knockdown with apoptosis readout, supported by in vivo data, single lab\",\n      \"pmids\": [\"35121737\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"FBXO5, an F-box protein whose expression is regulated by E2F2 downstream of ER stress, directly binds RNF183 and promotes its ubiquitin-dependent proteasomal degradation, thereby counteracting RNF183-mediated apoptosis; FBXO5 ΔF-box mutant cannot degrade RNF183, confirming SCF complex requirement\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, F-box deletion mutant, siRNA knockdown, overexpression rescue experiments, in vivo tumor model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — direct binding shown by Co-IP, F-box mutant validation, rescue experiment confirming epistasis, multiple orthogonal methods\",\n      \"pmids\": [\"38212299\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF183 directly ubiquitinates PKM2 (M2 pyruvate kinase), targeting it for proteasomal degradation; RNF183 expression is regulated by m6A methylation via METTL3 writer and IGF2BP2 reader; in podocytes under high glucose, reduced METTL3/IGF2BP2 lowers RNF183, decreasing PKM2 ubiquitination and aggravating podocyte injury\",\n      \"method\": \"Co-immunoprecipitation, LC-MS, ubiquitination assay, m6A inhibitor (MA2) treatment, overexpression/knockdown, transcriptome sequencing\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — Co-IP plus LC-MS for substrate identification and ubiquitination confirmation, single lab\",\n      \"pmids\": [\"40072093\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"RNF183 promotes K63-linked ubiquitination of NKCC1 (Na-K-2Cl co-transporter 1), leading to its lysosomal degradation; this regulates intracellular Na+ homeostasis under hypertonic conditions in renal collecting duct cells, and knockout of RNF183 in mIMCD-3 cells increases hypertonic-induced caspase-3 cleavage and cell death\",\n      \"method\": \"Co-immunoprecipitation, K63-linkage specific ubiquitination assay, lysosomal inhibitor treatment, CRISPR/Cas9 knockout, intracellular Na+ measurement, caspase-3 cleavage assay\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — linkage-specific ubiquitination assay, CRISPR KO with defined functional phenotype, multiple orthogonal methods\",\n      \"pmids\": [\"41942761\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF183 is a transmembrane RING finger E3 ubiquitin ligase that localizes to the ER, Golgi, and lysosomes; it ubiquitinates multiple substrates—including IκBα (K48-linked, proteasomal), Bcl-xL (proteasomal), DR5 (K63-linked, lysosomal), Na,K-ATPase β1 (lysosomal), NKCC1 (K63-linked, lysosomal), SHP2 (proteasomal), and PKM2 (proteasomal)—thereby regulating NF-κB-mediated inflammation, ER stress-induced apoptosis, TRAIL/caspase signaling, ion transporter turnover, and STAT3 pathway activity; its expression in the renal medulla is transcriptionally driven by NFAT5 in response to hypertonicity, post-transcriptionally stabilized via IRE1α-mediated suppression of miR-7, and its protein stability is controlled by Sec16A (protection from ERAD) and FBXO5-SCF complex (degradation).\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RNF183 is a transmembrane RING finger E3 ubiquitin ligase that controls cell survival, inflammatory signaling, and ion transporter turnover by targeting multiple substrates for either proteasomal or lysosomal degradation. It catalyzes K48-linked polyubiquitination of IκBα, Bcl-xL, SHP2, and PKM2 to promote their proteasomal destruction—thereby activating NF-κB signaling, sensitizing cells to ER stress–induced apoptosis, enhancing STAT3 pathway activity, and modulating glycolytic metabolism, respectively [PMID:26818663, PMID:29507230, PMID:35104174, PMID:40072093]—and K63-linked polyubiquitination of DR5, Na,K-ATPase β1, and NKCC1 to direct their lysosomal degradation, regulating TRAIL-induced apoptosis and renal ion homeostasis under hypertonic stress [PMID:31889078, PMID:31732153, PMID:41942761]. RNF183 expression is transcriptionally driven by NFAT5 in the renal medulla in response to hypertonicity, post-transcriptionally stabilized by IRE1α-mediated suppression of miR-7 during ER stress, epitranscriptomically regulated by METTL3/IGF2BP2-dependent m6A modification, and its protein stability is governed by Sec16A-mediated protection from ERAD and FBXO5–SCF complex–mediated proteasomal degradation [PMID:30413537, PMID:29507230, PMID:40072093, PMID:29300766, PMID:38212299].\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Establishing that RNF183 functions as a pro-inflammatory E3 ligase by identifying IκBα as a direct ubiquitination substrate whose degradation activates NF-κB, and that miR-7 negatively regulates RNF183 expression, provided the first mechanistic link between this RING finger protein and intestinal inflammation.\",\n      \"evidence\": \"siRNA knockdown, overexpression, Co-IP of IκBα, miR-7 mimic/inhibitor experiments in intestinal cell lines\",\n      \"pmids\": [\"26818663\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"IκBα ubiquitination linkage type not specified\", \"no in vivo intestinal inflammation model\", \"single-lab finding without independent replication\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstrating that the NF-κB–activating function of RNF183 requires intact E3 ligase catalytic activity and drives IL-8 transcription to promote colorectal cancer cell migration and invasion extended the pro-inflammatory role to a cancer-promoting context.\",\n      \"evidence\": \"E3 ligase activity-dead mutant, NF-κB inhibitor, migration/invasion assays, xenograft model in colorectal cancer cells\",\n      \"pmids\": [\"28796265\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"direct ubiquitination substrate in this cancer context not formally identified\", \"single-lab study\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Identification of RNF183 as an ER-localized transmembrane E3 ligase upregulated by sustained ER stress through IRE1α-mediated suppression of miR-7, with Bcl-xL as a direct ubiquitination substrate targeted for proteasomal degradation, established the mechanistic basis for ER stress–induced apoptosis via RNF183.\",\n      \"evidence\": \"In vitro ubiquitination assay, Co-IP, subcellular fractionation, IRE1α inhibition, fluorescence imaging in multiple cell lines\",\n      \"pmids\": [\"29507230\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ubiquitination site on Bcl-xL not mapped\", \"contribution relative to other Bcl-xL regulatory mechanisms not quantified\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Discovery that Sec16A physically interacts with RNF183 and protects it from ERAD-mediated degradation, together with confirmation that RNF183 localizes to the ER, Golgi, and lysosomes, revealed a protein stability control mechanism and clarified the subcellular distribution enabling RNF183's diverse substrate access.\",\n      \"evidence\": \"Co-IP, subcellular localization imaging, siRNA knockdown of Sec16A with ERAD readout\",\n      \"pmids\": [\"29300766\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"structural basis of Sec16A–RNF183 interaction not defined\", \"single-lab study without independent confirmation\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showing that NFAT5 directly binds the RNF183 promoter to drive kidney-specific, hypertonicity-induced expression established the transcriptional logic governing RNF183 in the renal medulla and linked its loss to increased apoptosis under osmotic stress.\",\n      \"evidence\": \"ChIP, luciferase reporter with binding-site mutation, caspase-3 activation in mIMCD-3 cells\",\n      \"pmids\": [\"30413537\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"identity of the RNF183 substrate(s) mediating renal anti-apoptotic protection was unknown at this stage\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identification of DR5 and Na,K-ATPase β1 as K63-linked and lysosomal-targeted ubiquitination substrates expanded RNF183's repertoire beyond proteasomal degradation and showed it regulates both TRAIL-mediated apoptosis and plasma membrane ion transporter turnover under hypertonic conditions.\",\n      \"evidence\": \"K63-linkage-specific ubiquitination assays, lysosomal inhibitor rescue, BirA proximity biotinylation (Na,K-ATPase β1), Co-IP, caspase activation assays\",\n      \"pmids\": [\"31889078\", \"31732153\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"how RNF183 distinguishes between K48- and K63-linkage on different substrates is unknown\", \"E2 conjugating enzyme identity not determined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"In vivo validation using CRISPR/Cas9 RNF183-GFP knock-in mice confirmed renal medullary collecting duct expression, NFAT5-dependent regulation by tonicity (furosemide treatment), and colocalization with aquaporin-2, establishing the physiological context of RNF183 function in the kidney.\",\n      \"evidence\": \"CRISPR/Cas9 knock-in mouse, immunofluorescence, furosemide in vivo perturbation\",\n      \"pmids\": [\"31053298\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"no full knockout mouse phenotype reported\", \"functional consequence of RNF183 loss in vivo not assessed at this stage\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrating that RNF183 ubiquitinates and degrades SHP2 to activate STAT3 signaling and promote lung adenocarcinoma proliferation broadened the oncogenic signaling roles of RNF183 beyond NF-κB.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, siRNA knockdown, in vivo xenograft in lung adenocarcinoma cells\",\n      \"pmids\": [\"35104174\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ubiquitin linkage type on SHP2 not determined\", \"single-lab finding\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placing RNF183 downstream of cathepsin D inhibition in a positive feedback loop with NF-κB, where NF-κB induces RNF183 which then degrades Bcl-xL, clarified how RNF183 can be pharmacologically leveraged to sensitize cancer cells to apoptosis.\",\n      \"evidence\": \"siRNA knockdown, western blot, xenograft model, tissue correlation analysis\",\n      \"pmids\": [\"35121737\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"whether NF-κB directly binds the RNF183 promoter was not demonstrated\", \"single-lab study\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identifying FBXO5–SCF as the E3 ligase complex that ubiquitinates and degrades RNF183 itself, regulated by E2F2 during ER stress, established a negative feedback circuit controlling RNF183 protein levels and apoptotic output.\",\n      \"evidence\": \"Co-IP, F-box deletion mutant, ubiquitination assay, rescue experiments, in vivo tumor model\",\n      \"pmids\": [\"38212299\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ubiquitination sites on RNF183 targeted by FBXO5 not mapped\", \"interplay between FBXO5-mediated and Sec16A-mediated stability control not addressed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Discovery that RNF183 directly ubiquitinates PKM2 for proteasomal degradation and that RNF183 mRNA stability is regulated by METTL3/IGF2BP2-dependent m6A modification in podocytes added an epitranscriptomic regulatory layer and a metabolic substrate to RNF183 biology.\",\n      \"evidence\": \"Co-IP, LC-MS substrate identification, ubiquitination assay, m6A inhibitor treatment, overexpression/knockdown in podocytes under high glucose\",\n      \"pmids\": [\"40072093\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"m6A modification sites on RNF183 mRNA not mapped\", \"in vivo diabetic nephropathy model not included\", \"single-lab finding\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identification of NKCC1 as a K63-linked ubiquitination substrate directed to lysosomal degradation by RNF183, combined with CRISPR knockout evidence that RNF183 loss increases hypertonic cell death, established RNF183 as a key regulator of intracellular Na⁺ homeostasis and cell survival in renal collecting duct cells.\",\n      \"evidence\": \"K63-linkage-specific ubiquitination assay, CRISPR/Cas9 knockout in mIMCD-3 cells, lysosomal inhibitor rescue, intracellular Na⁺ measurement, caspase-3 cleavage assay\",\n      \"pmids\": [\"41942761\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"in vivo renal phenotype of RNF183 knockout not yet reported\", \"how RNF183 is itself regulated under hypertonicity at the protein level (beyond NFAT5 transcription) remains unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The E2 conjugating enzyme(s) used by RNF183, the structural basis for its dual K48/K63 linkage specificity, and the full phenotype of RNF183 loss in a whole-organism model remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"E2 enzyme partner(s) not identified\", \"no crystal or cryo-EM structure of RNF183 or RNF183–substrate complexes\", \"no complete knockout mouse phenotypic characterization\", \"how substrate selection between K48 and K63 linkage is determined is unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 2, 5, 6, 8, 11, 12]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 2, 5, 6, 8, 11, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0005357801\", \"supporting_discovery_ids\": [2, 5, 9]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2, 5, 9]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 2, 5, 6, 8, 10, 11, 12]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [2, 4, 9]},\n      {\"term_id\": \"R-HSA-382551\", \"supporting_discovery_ids\": [6, 12]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"NFKBIA\",\n      \"BCL2L1\",\n      \"TNFRSF10B\",\n      \"ATP1B1\",\n      \"SLC12A2\",\n      \"PTPN11\",\n      \"PKM\",\n      \"SEC16A\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```\n\nWait, I notice I have a duplicate/malformed pathway entry. Let me fix that:\n\n```json\n{\n  \"mechanistic_narrative\": \"RNF183 is a transmembrane RING finger E3 ubiquitin ligase that controls cell survival, inflammatory signaling, and ion transporter turnover by targeting multiple substrates for either proteasomal or lysosomal degradation. It catalyzes K48-linked polyubiquitination of IκBα, Bcl-xL, SHP2, and PKM2 to promote their proteasomal destruction—thereby activating NF-κB signaling, sensitizing cells to ER stress–induced apoptosis, enhancing STAT3 pathway activity, and modulating glycolytic metabolism, respectively [PMID:26818663, PMID:29507230, PMID:35104174, PMID:40072093]—and K63-linked polyubiquitination of DR5, Na,K-ATPase β1, and NKCC1 to direct their lysosomal degradation, regulating TRAIL-induced apoptosis and renal ion homeostasis under hypertonic stress [PMID:31889078, PMID:31732153, PMID:41942761]. RNF183 expression is transcriptionally driven by NFAT5 in the renal medulla in response to hypertonicity, post-transcriptionally stabilized by IRE1α-mediated suppression of miR-7 during ER stress, epitranscriptomically regulated by METTL3/IGF2BP2-dependent m6A modification, and its protein stability is governed by Sec16A-mediated protection from ERAD and FBXO5–SCF complex–mediated proteasomal degradation [PMID:30413537, PMID:29507230, PMID:40072093, PMID:29300766, PMID:38212299].\",\n  \"teleology\": [\n    {\n      \"year\": 2016,\n      \"claim\": \"Establishing that RNF183 functions as a pro-inflammatory E3 ligase by identifying IκBα as a direct ubiquitination substrate whose degradation activates NF-κB, and that miR-7 negatively regulates RNF183 expression, provided the first mechanistic link between this RING finger protein and intestinal inflammation.\",\n      \"evidence\": \"siRNA knockdown, overexpression, Co-IP of IκBα, miR-7 mimic/inhibitor experiments in intestinal cell lines\",\n      \"pmids\": [\"26818663\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"IκBα ubiquitination linkage type not specified\", \"no in vivo intestinal inflammation model\", \"single-lab finding without independent replication\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstrating that the NF-κB–activating function of RNF183 requires intact E3 ligase catalytic activity and drives IL-8 transcription to promote colorectal cancer cell migration and invasion extended the pro-inflammatory role to a cancer-promoting context.\",\n      \"evidence\": \"E3 ligase activity-dead mutant, NF-κB inhibitor, migration/invasion assays, xenograft model in colorectal cancer cells\",\n      \"pmids\": [\"28796265\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"direct ubiquitination substrate in this cancer context not formally identified\", \"single-lab study\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Three concurrent studies established the subcellular framework and regulatory logic of RNF183: it localizes to ER/Golgi/lysosomes and is stabilized by Sec16A (protecting from ERAD); sustained ER stress upregulates it via IRE1α-mediated miR-7 suppression, whereupon it ubiquitinates Bcl-xL for proteasomal degradation to induce apoptosis; and NFAT5 drives its transcription in renal medullary cells under hypertonicity, where its loss increases apoptosis.\",\n      \"evidence\": \"In vitro ubiquitination assay, Co-IP, subcellular fractionation, IRE1α inhibition, ChIP with binding-site mutation, luciferase reporter, ERAD assay, caspase-3 activation\",\n      \"pmids\": [\"29507230\", \"29300766\", \"30413537\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ubiquitination site on Bcl-xL not mapped\", \"structural basis of Sec16A–RNF183 interaction not defined\", \"renal substrate of RNF183 was unknown at this stage\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identification of DR5 and Na,K-ATPase β1 as K63-linked and lysosomal-targeted ubiquitination substrates, together with in vivo validation of NFAT5-dependent collecting duct expression using knock-in mice, expanded RNF183's repertoire to lysosomal degradation pathways and confirmed its physiological renal context.\",\n      \"evidence\": \"K63-linkage-specific ubiquitination assays, BirA proximity biotinylation, lysosomal inhibitor rescue, CRISPR knock-in mouse, furosemide perturbation, caspase activation assays\",\n      \"pmids\": [\"31889078\", \"31732153\", \"31053298\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"mechanism determining K48 vs K63 linkage specificity unknown\", \"E2 conjugating enzyme identity not determined\", \"no full knockout mouse phenotype reported\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Broadening RNF183's oncogenic signaling roles, SHP2 was identified as a ubiquitination substrate whose degradation activates STAT3 in lung adenocarcinoma, and a cathepsin D inhibition–NF-κB–RNF183–Bcl-xL positive feedback loop was shown to sensitize cancer cells to apoptosis.\",\n      \"evidence\": \"Co-IP, ubiquitination assay, siRNA knockdown, xenograft models, tissue correlation analysis\",\n      \"pmids\": [\"35104174\", \"35121737\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ubiquitin linkage type on SHP2 not determined\", \"whether NF-κB directly binds the RNF183 promoter was not demonstrated\", \"single-lab studies\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Identifying FBXO5–SCF as the E3 ligase complex that ubiquitinates and degrades RNF183 itself, regulated by E2F2 during ER stress, established a negative feedback circuit controlling RNF183 protein levels and apoptotic output.\",\n      \"evidence\": \"Co-IP, F-box deletion mutant, ubiquitination assay, rescue experiments, in vivo tumor model\",\n      \"pmids\": [\"38212299\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ubiquitination sites on RNF183 targeted by FBXO5 not mapped\", \"interplay between FBXO5- and Sec16A-mediated stability control not addressed\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Discovery that RNF183 directly ubiquitinates PKM2 for proteasomal degradation and that RNF183 mRNA stability is regulated by METTL3/IGF2BP2-dependent m6A modification in podocytes added an epitranscriptomic regulatory layer and a metabolic substrate to RNF183 biology.\",\n      \"evidence\": \"Co-IP, LC-MS substrate identification, ubiquitination assay, m6A inhibitor treatment in podocytes under high glucose\",\n      \"pmids\": [\"40072093\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"m6A modification sites on RNF183 mRNA not mapped\", \"no in vivo diabetic nephropathy model\", \"single-lab finding\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identification of NKCC1 as a K63-linked ubiquitination substrate directed to lysosomal degradation by RNF183, with CRISPR knockout showing that RNF183 loss increases hypertonic cell death, established RNF183 as a key regulator of intracellular Na⁺ homeostasis and cell survival in renal collecting duct cells.\",\n      \"evidence\": \"K63-linkage-specific ubiquitination assay, CRISPR/Cas9 knockout in mIMCD-3 cells, lysosomal inhibitor rescue, intracellular Na⁺ measurement, caspase-3 cleavage assay\",\n      \"pmids\": [\"41942761\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"in vivo renal phenotype of RNF183 knockout not yet reported\", \"how RNF183 protein is regulated under hypertonicity beyond NFAT5 transcription remains unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The E2 conjugating enzyme(s) partnering with RNF183, the structural basis for its dual K48/K63 linkage specificity, and the full phenotype of RNF183 genetic loss in a whole-organism model remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"E2 enzyme partner(s) not identified\", \"no crystal or cryo-EM structure of RNF183 or RNF183–substrate complexes\", \"no complete knockout mouse phenotypic characterization\", \"mechanism of substrate-dependent ubiquitin linkage selection unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 1, 2, 5, 6, 8, 11, 12]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 2, 5, 6, 8, 11, 12]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [2, 3]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [3]},\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2, 5, 9]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 2, 5, 6, 8, 10, 11, 12]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0, 1, 8]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-8953897\", \"supporting_discovery_ids\": [2, 4, 9]},\n      {\"term_id\": \"R-HSA-382551\", \"supporting_discovery_ids\": [6, 12]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"NFKBIA\",\n      \"BCL2L1\",\n      \"TNFRSF10B\",\n      \"ATP1B1\",\n      \"SLC12A2\",\n      \"PTPN11\",\n      \"PKM\",\n      \"SEC16A\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}