{"gene":"RNF180","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2008,"finding":"Rines/RNF180 is an integral membrane E3 ubiquitin ligase localized to the endoplasmic reticulum membrane/nuclear envelope. It is not extracted by high salt or alkaline buffers and is degraded by proteinase K in intact ER, confirming it is an integral membrane protein with its N-terminal regions in the cytoplasm. It binds the UbcH6 ubiquitin-conjugating enzyme, promotes trans-ubiquitination, and its overexpression promotes proteasome-dependent degradation of a binding partner while Rines itself is heavily ubiquitinated and degraded by the proteasome.","method":"Subcellular fractionation, proteinase K protection assay, high salt/alkaline extraction, co-immunoprecipitation with UbcH6, proteasomal inhibitor treatment, trans-ubiquitination assay in cultured mammalian cells","journal":"Genes to cells : devoted to molecular & cellular mechanisms","confidence":"High","confidence_rationale":"Tier 1-2 / Strong — multiple orthogonal biochemical methods (fractionation, protease protection, alkaline extraction, co-IP, ubiquitination assay) in a single focused study establishing both localization and enzymatic activity","pmids":["18363970"],"is_preprint":false},{"year":2011,"finding":"RNF180 re-expression in gastric cancer cells suppresses cell growth and induces apoptosis, mediated by upregulation of MTSS1, CDKN2A, and TIMP3. The functional core promoter region (-202/+372) lies within a CpG island and can be silenced by in vitro methylation.","method":"Colony formation assay, annexin V apoptosis assay, cDNA microarray for target genes, 5' RACE for transcription start site, luciferase activity assay, in vitro methylation assay","journal":"Cancer","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — multiple functional readouts (growth, apoptosis, target gene array) with re-expression in cancer cell lines, single lab","pmids":["21717426"],"is_preprint":false},{"year":2016,"finding":"Specific CpG islands in the RNF180 promoter (CpG+102 and CpG+97) mediate the malignant biological characteristics of gastric cancer cells. Targeted demethylation of CpG+102 markedly influenced proliferation, invasion, cell cycle, anti-apoptosis, and tumorigenicity; CpG+97 was identified as an additional key locus.","method":"Site-specific demethylation via vector transfection in MGC-803 cells, proliferation assay, invasion assay, cell cycle analysis, apoptosis assay, tumorigenicity assay","journal":"Oncotarget","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple functional readouts with site-specific demethylation constructs, single lab","pmids":["27223257"],"is_preprint":false},{"year":2020,"finding":"RNF180 directly binds RhoC and promotes its ubiquitination and proteasomal degradation in gastric cancer cells. This reduces STAT3 phosphorylation downstream. RhoC knockdown phenocopies RNF180 overexpression in suppressing STAT3 phosphorylation, placing RhoC between RNF180 and STAT3.","method":"Co-immunoprecipitation (direct binding of RNF180 and RhoC), ubiquitin label-free quantitative proteomics, shRNA knockdown of RhoC, Western blot for STAT3 phosphorylation, transfection of RNF180-GFP expression vector","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP plus ubiquitin proteomics plus epistasis (RhoC KD rescues STAT3 phenotype), single lab","pmids":["33082325"],"is_preprint":false},{"year":2021,"finding":"RNF180 ubiquitinates DNMT3A and promotes its proteasome-mediated degradation. Loss of RNF180 allows DNMT3A to hypermethylate the ADAMTS9 promoter, silencing ADAMTS9 expression. This defines a RNF180/DNMT3A/ADAMTS9 regulatory axis in gastric cancer cells.","method":"Co-immunoprecipitation (RNF180-DNMT3A interaction), ubiquitination assay, proteasome inhibitor treatment, RNA-sequencing, promoter methylation analysis, cell viability and motility assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, ubiquitination assay, and downstream pathway (methylation/gene expression) validated, single lab","pmids":["33931579"],"is_preprint":false},{"year":2021,"finding":"RNF180 ubiquitinates WISP1, leading to WISP1 downregulation, which mediates RNF180's tumor-suppressive effects (inhibition of cell viability, induction of apoptosis) in colorectal cancer cells and in patient-derived xenograft mouse models.","method":"Co-immunoprecipitation, ubiquitination assay, overexpression/depletion of RNF180 and WISP1, cell viability assay, apoptosis assay, PDX mouse model","journal":"Frontiers in cell and developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, ubiquitination assay, in vivo PDX model, single lab","pmids":["33553163"],"is_preprint":false},{"year":2023,"finding":"RNF180 directly interacts with DNMT1 and promotes DNMT1 degradation via ubiquitination, preventing hypermethylation of the PCDH10 promoter and restoring PCDH10 expression in gastric cancer cells. This RNF180/DNMT1/PCDH10 axis suppresses GC cell proliferation and metastasis.","method":"Co-immunoprecipitation (RNF180-DNMT1 direct binding), ubiquitination assay, promoter methylation analysis, proliferation and metastasis assays, protein expression analysis in GC tissues","journal":"Clinical epigenetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, ubiquitination assay, and epigenetic downstream validation, single lab","pmids":["37147733"],"is_preprint":false},{"year":2023,"finding":"RNF180 interacts with IPO4 (Importin 4) and promotes its ubiquitination and proteasomal degradation in ovarian cancer cells. This reduces nuclear translocation of SOX2 (which requires IPO4 for nuclear import), increases p21 expression, and suppresses ovarian cancer progression.","method":"Bioinformatics, proteomics, co-immunoprecipitation (RNF180-IPO4), ubiquitination assay, IPO4 overexpression/knockdown, nuclear fractionation for SOX2 localization, in vitro and in vivo functional assays","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, ubiquitination assay, nuclear fractionation showing functional consequence, single lab","pmids":["37923100"],"is_preprint":false},{"year":2023,"finding":"BCL6 expression and tumor suppressor function in gastric cancer are regulated by the RNF180/RhoC pathway; specifically, RNF180-mediated ubiquitination and degradation of RhoC elevates BCL6 activity, which then transcriptionally represses FZD7 to inhibit Wnt signaling and promote ferroptosis via FZD7/β-catenin/TP63/GPX4 pathway.","method":"CHX chase and MG132 proteasome inhibitor treatment, rescue experiments, ChIP, dual luciferase reporter assay, lipid peroxidation/MDA/Fe2+ measurements, cell proliferation and metastasis assays","journal":"Cell & bioscience","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (CHX, MG132, ChIP, luciferase, ferroptosis assays), single lab","pmids":["37060074"],"is_preprint":false},{"year":2022,"finding":"RNF180 directly interacts with RAD51 and promotes its ubiquitination and degradation in triple-negative breast cancer cells, thereby suppressing TNBC proliferation, invasion, migration, and sensitizing cells to Gefitinib.","method":"Immunoprecipitation (RNF180-RAD51 interaction), ubiquitination assay, gain- and loss-of-function assays, Western blot for drug resistance genes, nude mouse xenograft model","journal":"Chemico-biological interactions","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP, ubiquitination assay, in vivo xenograft, single lab","pmids":["34998818"],"is_preprint":false},{"year":2024,"finding":"RNF180 promotes ubiquitination and degradation of ALKBH5 (an m6A RNA demethylase) in the context of ulcerative colitis. ALKBH5 normally inhibits SMARCA5 via m6A modification; loss of ALKBH5 due to RNF180-mediated degradation increases SMARCA5 and worsens colon inflammation and Th17/Treg imbalance.","method":"UC mouse model (DSS-induced), RNF180/SMARCA5 knockdown, ALKBH5 overexpression, flow cytometry (Th17/Treg cells), cytokine measurements, Western blot; ubiquitination of ALKBH5 by RNF180 established","journal":"Archives of pharmacal research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo mouse model with knockdown/OE, functional cytokine and immune readouts, ubiquitination mechanism described, single lab","pmids":["39060657"],"is_preprint":false},{"year":2025,"finding":"RNF180 ubiquitinates and promotes proteasomal degradation of ACC1 and ACLY in esophageal cancer cells, thereby weakening lipid droplet formation and lipogenesis, which reduces chemoresistance.","method":"RNF180 overexpression in EC cell lines, ubiquitination and proteasomal degradation assays for ACC1 and ACLY, lipid droplet measurement, chemosensitivity assays, in vitro proliferation/migration/invasion/apoptosis/pyroptosis assays","journal":"Frontiers in pharmacology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — ubiquitination mechanism described but limited methodological detail in abstract, single lab, single study","pmids":["40331188"],"is_preprint":false},{"year":2025,"finding":"RNF180 interacts with and promotes ubiquitination and proteasomal degradation of NOTCH1, TRIM24, and FOXC1 in colorectal cancer cells, suppressing aggressiveness via Akt pathway inactivation. RNF180 also ubiquitinates and degrades ACC1 and ACLY to suppress lipid droplet assembly and chemoresistance. RNF180 knockout in Lgr5-cre/RNF180 f/f mice enhanced sensitivity to chemically-induced colorectal carcinogenesis.","method":"Co-immunoprecipitation, ubiquitination and proteasomal degradation assays, Akt pathway analysis, lipid droplet assay, conditional knockout mouse model (Lgr5-cre/RNF180 f/f), in vivo and in vitro functional assays","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, ubiquitination assay, conditional KO mouse model with in vivo phenotype, multiple substrates identified, single lab","pmids":["40915184"],"is_preprint":false},{"year":2026,"finding":"RNF180 ubiquitinates and degrades PLK2 in non-small cell lung cancer cells, suppressing EGFR/PI3K/AKT signaling and reducing cisplatin resistance. RNF180 overexpression inhibits proliferation, migration, invasion, EMT, and drug resistance protein expression while promoting apoptosis.","method":"Bioinformatics identification of PLK2 as substrate, qRT-PCR, Western blot, lentiviral RNF180 overexpression, PLK2 knockdown, nude mouse xenograft model, signaling pathway analysis","journal":"Scientific reports","confidence":"Low","confidence_rationale":"Tier 3 / Weak — substrate identified by bioinformatics with partial validation, limited mechanistic detail in abstract, single lab","pmids":["41807497"],"is_preprint":false}],"current_model":"RNF180 is an integral membrane E3 ubiquitin ligase resident in the ER/nuclear envelope that binds E2 ubiquitin-conjugating enzyme UbcH6 and directly ubiquitinates multiple substrates—including RhoC, DNMT1, DNMT3A, WISP1, IPO4, RAD51, ALKBH5, ACC1, ACLY, NOTCH1, TRIM24, FOXC1, and PLK2—targeting them for proteasomal degradation, thereby suppressing tumor cell proliferation, invasion, and chemoresistance across gastric, colorectal, ovarian, breast, esophageal, and lung cancers, in part by regulating downstream transcription factors (STAT3, SOX2), DNA methylation-dependent gene silencing (via DNMT1/DNMT3A), lipid metabolism, and the EGFR/PI3K/AKT pathway."},"narrative":{"mechanistic_narrative":"RNF180 (Rines) is an integral membrane E3 ubiquitin ligase resident in the endoplasmic reticulum membrane and nuclear envelope, with its N-terminal regions exposed to the cytoplasm, that binds the E2 ubiquitin-conjugating enzyme UbcH6 to catalyze trans-ubiquitination and direct substrates to proteasomal degradation [PMID:18363970]. Across multiple epithelial cancers it acts as a tumor suppressor by ubiquitinating and degrading a range of pro-tumorigenic proteins, restraining proliferation, invasion, and chemoresistance [PMID:33082325, PMID:33553163, PMID:40915184]. A recurrent theme is RNF180-mediated control of the DNA methylation machinery: it directly degrades DNMT1 and DNMT3A, preventing hypermethylation and silencing of tumor-suppressor genes such as PCDH10 and ADAMTS9 [PMID:33931579, PMID:37147733]. It also degrades RhoC to dampen STAT3 phosphorylation and engage a BCL6/FZD7 axis controlling Wnt signaling and ferroptosis [PMID:33082325, PMID:37060074], and degrades IPO4 to block nuclear import of SOX2 [PMID:37923100]. Additional substrates link RNF180 to homologous recombination (RAD51) [PMID:34998818], lipid metabolism through ACC1 and ACLY [PMID:40915184], and Akt/EGFR signaling via NOTCH1, TRIM24, FOXC1, and PLK2 [PMID:40915184, PMID:41807497]. RNF180 itself is frequently silenced in gastric cancer by promoter CpG-island methylation, and restoring its expression suppresses growth and induces apoptosis [PMID:21717426, PMID:27223257]. Conditional Rnf180 knockout in mice increases susceptibility to chemically induced colorectal carcinogenesis, supporting an in vivo tumor-suppressive role [PMID:40915184].","teleology":[{"year":2008,"claim":"Established the fundamental biochemical identity of RNF180 — answering whether it is an active enzyme and where it acts — by showing it is an ER/nuclear-envelope integral membrane E3 ligase that uses UbcH6 and drives proteasomal degradation of partners.","evidence":"Subcellular fractionation, proteinase K protection, alkaline extraction, UbcH6 co-IP, and trans-ubiquitination assays in mammalian cells","pmids":["18363970"],"confidence":"High","gaps":["Physiological substrates not identified at this stage","Catalytic domain/RING architecture not structurally resolved","Role in cancer not yet addressed"]},{"year":2011,"claim":"Linked RNF180 to tumor suppression and to its own epigenetic silencing, showing re-expression suppresses gastric cancer growth and that its CpG-island promoter is methylation-sensitive.","evidence":"Colony formation, apoptosis assays, cDNA microarray, 5'RACE, luciferase and in vitro methylation assays in gastric cancer cells","pmids":["21717426"],"confidence":"Medium","gaps":["MTSS1/CDKN2A/TIMP3 identified as correlated targets but not as direct ubiquitination substrates","Mechanism connecting ligase activity to these outputs unresolved"]},{"year":2016,"claim":"Refined the epigenetic-silencing model by mapping specific CpG loci (CpG+102, CpG+97) whose methylation controls RNF180's tumor-suppressive output.","evidence":"Site-specific demethylation constructs with proliferation, invasion, cell cycle, apoptosis, and tumorigenicity assays in MGC-803 cells","pmids":["27223257"],"confidence":"Medium","gaps":["Causal link between specific CpG demethylation and downstream substrate ubiquitination not established","Single cell line"]},{"year":2020,"claim":"Identified the first defined oncogenic substrate, showing RNF180 degrades RhoC to suppress STAT3 phosphorylation, establishing a RNF180→RhoC→STAT3 epistatic axis.","evidence":"Co-IP, ubiquitin label-free proteomics, RhoC shRNA rescue, and STAT3 phospho-Western in gastric cancer cells","pmids":["33082325"],"confidence":"Medium","gaps":["Direct in vitro ubiquitination with purified components not shown","Single lab"]},{"year":2021,"claim":"Connected RNF180 to control of the DNA methylation machinery and to a second cancer type, showing it degrades DNMT3A to prevent ADAMTS9 silencing and degrades WISP1 to suppress colorectal tumor growth in PDX models.","evidence":"Co-IP, ubiquitination assays, RNA-seq, promoter methylation analysis, and PDX mouse models","pmids":["33931579","33553163"],"confidence":"Medium","gaps":["Whether DNMT3A and WISP1 are co-regulated in the same cells unclear","Substrate-recognition determinants unknown"]},{"year":2022,"claim":"Extended substrate range to DNA-repair machinery, showing RNF180 degrades RAD51 in triple-negative breast cancer and sensitizes cells to Gefitinib.","evidence":"IP, ubiquitination assay, gain/loss-of-function studies, and nude mouse xenograft","pmids":["34998818"],"confidence":"Medium","gaps":["Impact on homologous recombination capacity not directly measured","Single lab"]},{"year":2023,"claim":"Consolidated RNF180 as a master regulator of methylation and transcription-factor trafficking, adding DNMT1 (PCDH10 axis), IPO4-dependent SOX2 nuclear import, and a RhoC/BCL6/FZD7 ferroptosis/Wnt branch.","evidence":"Co-IP, ubiquitination assays, nuclear fractionation, ChIP, dual luciferase, and ferroptosis/lipid-peroxidation assays across gastric and ovarian cancer cells","pmids":["37147733","37923100","37060074"],"confidence":"Medium","gaps":["Whether these parallel axes operate simultaneously or are context-specific is unresolved","Direct ubiquitination chain-type/specificity uncharacterized"]},{"year":2024,"claim":"Demonstrated a non-cancer, immunological role by showing RNF180 degrades the m6A demethylase ALKBH5, raising SMARCA5 and worsening colitis with Th17/Treg imbalance.","evidence":"DSS-induced ulcerative colitis mouse model with knockdown/overexpression, flow cytometry, cytokine measurements, and ubiquitination analysis","pmids":["39060657"],"confidence":"Medium","gaps":["Whether the same RNF180 activity is pro- or anti-disease depends on context and is unresolved","Single lab"]},{"year":2025,"claim":"Provided in vivo genetic support and broadened substrate scope into metabolism and Akt signaling, with conditional knockout enhancing colorectal carcinogenesis and degradation of NOTCH1/TRIM24/FOXC1, ACC1, and ACLY linking RNF180 to lipid metabolism and chemoresistance.","evidence":"Co-IP, ubiquitination/degradation assays, lipid droplet assays, Akt pathway analysis, and Lgr5-cre/RNF180 f/f conditional knockout mouse","pmids":["40915184","40331188"],"confidence":"Medium","gaps":["ACC1/ACLY findings (idx 11) rest on limited methodological detail","Whether all listed substrates are direct in vitro substrates not uniformly established"]},{"year":2026,"claim":"Added PLK2 as a substrate linking RNF180 to EGFR/PI3K/AKT signaling and cisplatin resistance in lung cancer.","evidence":"Bioinformatic substrate prediction with qRT-PCR, Western blot, lentiviral overexpression, PLK2 knockdown, and nude mouse xenograft","pmids":["41807497"],"confidence":"Low","gaps":["Substrate identified largely by bioinformatics with partial validation","Direct ubiquitination not rigorously demonstrated","Single lab"]},{"year":null,"claim":"It remains unknown how RNF180 achieves specificity across its very broad reported substrate set and which substrates are physiologically dominant in each tissue context.","evidence":"No reconstituted system defining recognition determinants or ubiquitin chain topology across substrates is present in the corpus","pmids":[],"confidence":"Low","gaps":["No structural basis for substrate selection","Degron/recognition motifs unmapped","Chain-type specificity of ubiquitination unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,3,4,6]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0]}],"localization":[{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[0]},{"term_id":"GO:0005635","term_label":"nuclear envelope","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0]}],"complexes":[],"partners":["UBCH6","RHOC","DNMT1","DNMT3A","WISP1","IPO4","RAD51","ALKBH5"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q86T96","full_name":"E3 ubiquitin-protein ligase RNF180","aliases":["RING finger protein 180","RING-type E3 ubiquitin transferase RNF180"],"length_aa":592,"mass_kda":68.3,"function":"E3 ubiquitin-protein ligase which promotes polyubiquitination and degradation by the proteasome pathway of ZIC2","subcellular_location":"Endoplasmic reticulum membrane; Nucleus envelope","url":"https://www.uniprot.org/uniprotkb/Q86T96/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF180","classification":"Not Classified","n_dependent_lines":5,"n_total_lines":1208,"dependency_fraction":0.0041390728476821195},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF180","total_profiled":1310},"omim":[{"mim_id":"616015","title":"RING FINGER PROTEIN 180; RNF180","url":"https://www.omim.org/entry/616015"},{"mim_id":"603073","title":"ZIC FAMILY, MEMBER 2; ZIC2","url":"https://www.omim.org/entry/603073"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"parathyroid gland","ntpm":23.8}],"url":"https://www.proteinatlas.org/search/RNF180"},"hgnc":{"alias_symbol":[],"prev_symbol":[]},"alphafold":{"accession":"Q86T96","domains":[{"cath_id":"-","chopping":"19-38_59-125","consensus_level":"high","plddt":85.0203,"start":19,"end":125},{"cath_id":"3.30.40.10","chopping":"430-513","consensus_level":"high","plddt":89.0069,"start":430,"end":513}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86T96","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q86T96-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q86T96-F1-predicted_aligned_error_v6.png","plddt_mean":56.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF180","jax_strain_url":"https://www.jax.org/strain/search?query=RNF180"},"sequence":{"accession":"Q86T96","fasta_url":"https://rest.uniprot.org/uniprotkb/Q86T96.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q86T96/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q86T96"}},"corpus_meta":[{"pmid":"21717426","id":"PMC_21717426","title":"Characterization of the gene structure, functional significance, and clinical application of RNF180, a novel gene in gastric cancer.","date":"2011","source":"Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/21717426","citation_count":71,"is_preprint":false},{"pmid":"18363970","id":"PMC_18363970","title":"Rines/RNF180, a novel RING finger gene-encoded product, is a membrane-bound ubiquitin ligase.","date":"2008","source":"Genes to cells : devoted to molecular & cellular mechanisms","url":"https://pubmed.ncbi.nlm.nih.gov/18363970","citation_count":41,"is_preprint":false},{"pmid":"25202403","id":"PMC_25202403","title":"Detection of aberrant promoter methylation of RNF180, DAPK1 and SFRP2 in plasma DNA of patients with gastric cancer.","date":"2014","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/25202403","citation_count":29,"is_preprint":false},{"pmid":"33082325","id":"PMC_33082325","title":"RNF180 mediates STAT3 activity by regulating the expression of RhoC via the proteasomal pathway in gastric cancer cells.","date":"2020","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/33082325","citation_count":28,"is_preprint":false},{"pmid":"33931579","id":"PMC_33931579","title":"DNMT3A-mediated silence in ADAMTS9 expression is restored by RNF180 to inhibit viability and motility in gastric cancer cells.","date":"2021","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/33931579","citation_count":25,"is_preprint":false},{"pmid":"27050149","id":"PMC_27050149","title":"Promoter methylation of RNF180 is associated with H.pylori infection and serves as a marker for gastric cancer and atrophic gastritis.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27050149","citation_count":20,"is_preprint":false},{"pmid":"37060074","id":"PMC_37060074","title":"The malignancy suppression and ferroptosis facilitation of BCL6 in gastric cancer mediated by FZD7 repression are strengthened by RNF180/RhoC 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of E3 ubiquitin ligase RNF180 DNA promoter to predict the survival of gastric cancer.","date":"2015","source":"Cancer biomarkers : section A of Disease markers","url":"https://pubmed.ncbi.nlm.nih.gov/25769451","citation_count":14,"is_preprint":false},{"pmid":"27223257","id":"PMC_27223257","title":"Mediation of the malignant biological characteristics of gastric cancer cells by the methylated CpG islands in RNF180 DNA promoter.","date":"2016","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/27223257","citation_count":10,"is_preprint":false},{"pmid":"33553163","id":"PMC_33553163","title":"RNF180 Inhibits Proliferation and Promotes Apoptosis of Colorectal Cancer Through Ubiquitination of WISP1.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/33553163","citation_count":8,"is_preprint":false},{"pmid":"37923100","id":"PMC_37923100","title":"E3 ubiquitin ligase RNF180 impairs IPO4/SOX2 complex stability and inhibits SOX2-mediated malignancy in ovarian cancer.","date":"2023","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/37923100","citation_count":7,"is_preprint":false},{"pmid":"36246966","id":"PMC_36246966","title":"Plasma Methylated RNF180 for Noninvasive Diagnosis of Gastric Cancer.","date":"2022","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/36246966","citation_count":7,"is_preprint":false},{"pmid":"34998818","id":"PMC_34998818","title":"E3 ubiquitin ligase RNF180 reduces sensitivity of triple-negative breast cancer cells to Gefitinib by downregulating RAD51.","date":"2022","source":"Chemico-biological interactions","url":"https://pubmed.ncbi.nlm.nih.gov/34998818","citation_count":5,"is_preprint":false},{"pmid":"39541711","id":"PMC_39541711","title":"Combining methylated RNF180 and SFRP2 plasma biomarkers for noninvasive diagnosis of gastric cancer.","date":"2024","source":"Translational 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immunopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/40915184","citation_count":0,"is_preprint":false},{"pmid":"41807497","id":"PMC_41807497","title":"The E3 ubiquitin ligase RNF180 modulates the EGFR/PI3K/AKT pathway to reduce cisplatin resistance in non-small cell lung cancer.","date":"2026","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/41807497","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13222,"output_tokens":3899,"usd":0.049076,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11590,"output_tokens":3639,"usd":0.074463,"stage2_stop_reason":"end_turn"},"total_usd":0.123539,"stage1_batch_id":"msgbatch_01CNoDbkChHEkgrJnL7BwtGJ","stage2_batch_id":"msgbatch_016Y4x9E9Wg5VQDANj9quSQD","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2008,\n      \"finding\": \"Rines/RNF180 is an integral membrane E3 ubiquitin ligase localized to the endoplasmic reticulum membrane/nuclear envelope. It is not extracted by high salt or alkaline buffers and is degraded by proteinase K in intact ER, confirming it is an integral membrane protein with its N-terminal regions in the cytoplasm. It binds the UbcH6 ubiquitin-conjugating enzyme, promotes trans-ubiquitination, and its overexpression promotes proteasome-dependent degradation of a binding partner while Rines itself is heavily ubiquitinated and degraded by the proteasome.\",\n      \"method\": \"Subcellular fractionation, proteinase K protection assay, high salt/alkaline extraction, co-immunoprecipitation with UbcH6, proteasomal inhibitor treatment, trans-ubiquitination assay in cultured mammalian cells\",\n      \"journal\": \"Genes to cells : devoted to molecular & cellular mechanisms\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Strong — multiple orthogonal biochemical methods (fractionation, protease protection, alkaline extraction, co-IP, ubiquitination assay) in a single focused study establishing both localization and enzymatic activity\",\n      \"pmids\": [\"18363970\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"RNF180 re-expression in gastric cancer cells suppresses cell growth and induces apoptosis, mediated by upregulation of MTSS1, CDKN2A, and TIMP3. The functional core promoter region (-202/+372) lies within a CpG island and can be silenced by in vitro methylation.\",\n      \"method\": \"Colony formation assay, annexin V apoptosis assay, cDNA microarray for target genes, 5' RACE for transcription start site, luciferase activity assay, in vitro methylation assay\",\n      \"journal\": \"Cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — multiple functional readouts (growth, apoptosis, target gene array) with re-expression in cancer cell lines, single lab\",\n      \"pmids\": [\"21717426\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Specific CpG islands in the RNF180 promoter (CpG+102 and CpG+97) mediate the malignant biological characteristics of gastric cancer cells. Targeted demethylation of CpG+102 markedly influenced proliferation, invasion, cell cycle, anti-apoptosis, and tumorigenicity; CpG+97 was identified as an additional key locus.\",\n      \"method\": \"Site-specific demethylation via vector transfection in MGC-803 cells, proliferation assay, invasion assay, cell cycle analysis, apoptosis assay, tumorigenicity assay\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple functional readouts with site-specific demethylation constructs, single lab\",\n      \"pmids\": [\"27223257\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF180 directly binds RhoC and promotes its ubiquitination and proteasomal degradation in gastric cancer cells. This reduces STAT3 phosphorylation downstream. RhoC knockdown phenocopies RNF180 overexpression in suppressing STAT3 phosphorylation, placing RhoC between RNF180 and STAT3.\",\n      \"method\": \"Co-immunoprecipitation (direct binding of RNF180 and RhoC), ubiquitin label-free quantitative proteomics, shRNA knockdown of RhoC, Western blot for STAT3 phosphorylation, transfection of RNF180-GFP expression vector\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP plus ubiquitin proteomics plus epistasis (RhoC KD rescues STAT3 phenotype), single lab\",\n      \"pmids\": [\"33082325\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF180 ubiquitinates DNMT3A and promotes its proteasome-mediated degradation. Loss of RNF180 allows DNMT3A to hypermethylate the ADAMTS9 promoter, silencing ADAMTS9 expression. This defines a RNF180/DNMT3A/ADAMTS9 regulatory axis in gastric cancer cells.\",\n      \"method\": \"Co-immunoprecipitation (RNF180-DNMT3A interaction), ubiquitination assay, proteasome inhibitor treatment, RNA-sequencing, promoter methylation analysis, cell viability and motility assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, ubiquitination assay, and downstream pathway (methylation/gene expression) validated, single lab\",\n      \"pmids\": [\"33931579\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF180 ubiquitinates WISP1, leading to WISP1 downregulation, which mediates RNF180's tumor-suppressive effects (inhibition of cell viability, induction of apoptosis) in colorectal cancer cells and in patient-derived xenograft mouse models.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, overexpression/depletion of RNF180 and WISP1, cell viability assay, apoptosis assay, PDX mouse model\",\n      \"journal\": \"Frontiers in cell and developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, ubiquitination assay, in vivo PDX model, single lab\",\n      \"pmids\": [\"33553163\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF180 directly interacts with DNMT1 and promotes DNMT1 degradation via ubiquitination, preventing hypermethylation of the PCDH10 promoter and restoring PCDH10 expression in gastric cancer cells. This RNF180/DNMT1/PCDH10 axis suppresses GC cell proliferation and metastasis.\",\n      \"method\": \"Co-immunoprecipitation (RNF180-DNMT1 direct binding), ubiquitination assay, promoter methylation analysis, proliferation and metastasis assays, protein expression analysis in GC tissues\",\n      \"journal\": \"Clinical epigenetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, ubiquitination assay, and epigenetic downstream validation, single lab\",\n      \"pmids\": [\"37147733\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF180 interacts with IPO4 (Importin 4) and promotes its ubiquitination and proteasomal degradation in ovarian cancer cells. This reduces nuclear translocation of SOX2 (which requires IPO4 for nuclear import), increases p21 expression, and suppresses ovarian cancer progression.\",\n      \"method\": \"Bioinformatics, proteomics, co-immunoprecipitation (RNF180-IPO4), ubiquitination assay, IPO4 overexpression/knockdown, nuclear fractionation for SOX2 localization, in vitro and in vivo functional assays\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, ubiquitination assay, nuclear fractionation showing functional consequence, single lab\",\n      \"pmids\": [\"37923100\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"BCL6 expression and tumor suppressor function in gastric cancer are regulated by the RNF180/RhoC pathway; specifically, RNF180-mediated ubiquitination and degradation of RhoC elevates BCL6 activity, which then transcriptionally represses FZD7 to inhibit Wnt signaling and promote ferroptosis via FZD7/β-catenin/TP63/GPX4 pathway.\",\n      \"method\": \"CHX chase and MG132 proteasome inhibitor treatment, rescue experiments, ChIP, dual luciferase reporter assay, lipid peroxidation/MDA/Fe2+ measurements, cell proliferation and metastasis assays\",\n      \"journal\": \"Cell & bioscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (CHX, MG132, ChIP, luciferase, ferroptosis assays), single lab\",\n      \"pmids\": [\"37060074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF180 directly interacts with RAD51 and promotes its ubiquitination and degradation in triple-negative breast cancer cells, thereby suppressing TNBC proliferation, invasion, migration, and sensitizing cells to Gefitinib.\",\n      \"method\": \"Immunoprecipitation (RNF180-RAD51 interaction), ubiquitination assay, gain- and loss-of-function assays, Western blot for drug resistance genes, nude mouse xenograft model\",\n      \"journal\": \"Chemico-biological interactions\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP, ubiquitination assay, in vivo xenograft, single lab\",\n      \"pmids\": [\"34998818\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF180 promotes ubiquitination and degradation of ALKBH5 (an m6A RNA demethylase) in the context of ulcerative colitis. ALKBH5 normally inhibits SMARCA5 via m6A modification; loss of ALKBH5 due to RNF180-mediated degradation increases SMARCA5 and worsens colon inflammation and Th17/Treg imbalance.\",\n      \"method\": \"UC mouse model (DSS-induced), RNF180/SMARCA5 knockdown, ALKBH5 overexpression, flow cytometry (Th17/Treg cells), cytokine measurements, Western blot; ubiquitination of ALKBH5 by RNF180 established\",\n      \"journal\": \"Archives of pharmacal research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo mouse model with knockdown/OE, functional cytokine and immune readouts, ubiquitination mechanism described, single lab\",\n      \"pmids\": [\"39060657\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF180 ubiquitinates and promotes proteasomal degradation of ACC1 and ACLY in esophageal cancer cells, thereby weakening lipid droplet formation and lipogenesis, which reduces chemoresistance.\",\n      \"method\": \"RNF180 overexpression in EC cell lines, ubiquitination and proteasomal degradation assays for ACC1 and ACLY, lipid droplet measurement, chemosensitivity assays, in vitro proliferation/migration/invasion/apoptosis/pyroptosis assays\",\n      \"journal\": \"Frontiers in pharmacology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — ubiquitination mechanism described but limited methodological detail in abstract, single lab, single study\",\n      \"pmids\": [\"40331188\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF180 interacts with and promotes ubiquitination and proteasomal degradation of NOTCH1, TRIM24, and FOXC1 in colorectal cancer cells, suppressing aggressiveness via Akt pathway inactivation. RNF180 also ubiquitinates and degrades ACC1 and ACLY to suppress lipid droplet assembly and chemoresistance. RNF180 knockout in Lgr5-cre/RNF180 f/f mice enhanced sensitivity to chemically-induced colorectal carcinogenesis.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination and proteasomal degradation assays, Akt pathway analysis, lipid droplet assay, conditional knockout mouse model (Lgr5-cre/RNF180 f/f), in vivo and in vitro functional assays\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, ubiquitination assay, conditional KO mouse model with in vivo phenotype, multiple substrates identified, single lab\",\n      \"pmids\": [\"40915184\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"RNF180 ubiquitinates and degrades PLK2 in non-small cell lung cancer cells, suppressing EGFR/PI3K/AKT signaling and reducing cisplatin resistance. RNF180 overexpression inhibits proliferation, migration, invasion, EMT, and drug resistance protein expression while promoting apoptosis.\",\n      \"method\": \"Bioinformatics identification of PLK2 as substrate, qRT-PCR, Western blot, lentiviral RNF180 overexpression, PLK2 knockdown, nude mouse xenograft model, signaling pathway analysis\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — substrate identified by bioinformatics with partial validation, limited mechanistic detail in abstract, single lab\",\n      \"pmids\": [\"41807497\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF180 is an integral membrane E3 ubiquitin ligase resident in the ER/nuclear envelope that binds E2 ubiquitin-conjugating enzyme UbcH6 and directly ubiquitinates multiple substrates—including RhoC, DNMT1, DNMT3A, WISP1, IPO4, RAD51, ALKBH5, ACC1, ACLY, NOTCH1, TRIM24, FOXC1, and PLK2—targeting them for proteasomal degradation, thereby suppressing tumor cell proliferation, invasion, and chemoresistance across gastric, colorectal, ovarian, breast, esophageal, and lung cancers, in part by regulating downstream transcription factors (STAT3, SOX2), DNA methylation-dependent gene silencing (via DNMT1/DNMT3A), lipid metabolism, and the EGFR/PI3K/AKT pathway.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RNF180 (Rines) is an integral membrane E3 ubiquitin ligase resident in the endoplasmic reticulum membrane and nuclear envelope, with its N-terminal regions exposed to the cytoplasm, that binds the E2 ubiquitin-conjugating enzyme UbcH6 to catalyze trans-ubiquitination and direct substrates to proteasomal degradation [#0]. Across multiple epithelial cancers it acts as a tumor suppressor by ubiquitinating and degrading a range of pro-tumorigenic proteins, restraining proliferation, invasion, and chemoresistance [#3, #5, #12]. A recurrent theme is RNF180-mediated control of the DNA methylation machinery: it directly degrades DNMT1 and DNMT3A, preventing hypermethylation and silencing of tumor-suppressor genes such as PCDH10 and ADAMTS9 [#4, #6]. It also degrades RhoC to dampen STAT3 phosphorylation and engage a BCL6/FZD7 axis controlling Wnt signaling and ferroptosis [#3, #8], and degrades IPO4 to block nuclear import of SOX2 [#7]. Additional substrates link RNF180 to homologous recombination (RAD51) [#9], lipid metabolism through ACC1 and ACLY [#12], and Akt/EGFR signaling via NOTCH1, TRIM24, FOXC1, and PLK2 [#12, #13]. RNF180 itself is frequently silenced in gastric cancer by promoter CpG-island methylation, and restoring its expression suppresses growth and induces apoptosis [#1, #2]. Conditional Rnf180 knockout in mice increases susceptibility to chemically induced colorectal carcinogenesis, supporting an in vivo tumor-suppressive role [#12].\",\n  \"teleology\": [\n    {\n      \"year\": 2008,\n      \"claim\": \"Established the fundamental biochemical identity of RNF180 — answering whether it is an active enzyme and where it acts — by showing it is an ER/nuclear-envelope integral membrane E3 ligase that uses UbcH6 and drives proteasomal degradation of partners.\",\n      \"evidence\": \"Subcellular fractionation, proteinase K protection, alkaline extraction, UbcH6 co-IP, and trans-ubiquitination assays in mammalian cells\",\n      \"pmids\": [\"18363970\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological substrates not identified at this stage\", \"Catalytic domain/RING architecture not structurally resolved\", \"Role in cancer not yet addressed\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Linked RNF180 to tumor suppression and to its own epigenetic silencing, showing re-expression suppresses gastric cancer growth and that its CpG-island promoter is methylation-sensitive.\",\n      \"evidence\": \"Colony formation, apoptosis assays, cDNA microarray, 5'RACE, luciferase and in vitro methylation assays in gastric cancer cells\",\n      \"pmids\": [\"21717426\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"MTSS1/CDKN2A/TIMP3 identified as correlated targets but not as direct ubiquitination substrates\", \"Mechanism connecting ligase activity to these outputs unresolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Refined the epigenetic-silencing model by mapping specific CpG loci (CpG+102, CpG+97) whose methylation controls RNF180's tumor-suppressive output.\",\n      \"evidence\": \"Site-specific demethylation constructs with proliferation, invasion, cell cycle, apoptosis, and tumorigenicity assays in MGC-803 cells\",\n      \"pmids\": [\"27223257\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Causal link between specific CpG demethylation and downstream substrate ubiquitination not established\", \"Single cell line\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified the first defined oncogenic substrate, showing RNF180 degrades RhoC to suppress STAT3 phosphorylation, establishing a RNF180→RhoC→STAT3 epistatic axis.\",\n      \"evidence\": \"Co-IP, ubiquitin label-free proteomics, RhoC shRNA rescue, and STAT3 phospho-Western in gastric cancer cells\",\n      \"pmids\": [\"33082325\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct in vitro ubiquitination with purified components not shown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connected RNF180 to control of the DNA methylation machinery and to a second cancer type, showing it degrades DNMT3A to prevent ADAMTS9 silencing and degrades WISP1 to suppress colorectal tumor growth in PDX models.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, RNA-seq, promoter methylation analysis, and PDX mouse models\",\n      \"pmids\": [\"33931579\", \"33553163\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether DNMT3A and WISP1 are co-regulated in the same cells unclear\", \"Substrate-recognition determinants unknown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Extended substrate range to DNA-repair machinery, showing RNF180 degrades RAD51 in triple-negative breast cancer and sensitizes cells to Gefitinib.\",\n      \"evidence\": \"IP, ubiquitination assay, gain/loss-of-function studies, and nude mouse xenograft\",\n      \"pmids\": [\"34998818\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Impact on homologous recombination capacity not directly measured\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Consolidated RNF180 as a master regulator of methylation and transcription-factor trafficking, adding DNMT1 (PCDH10 axis), IPO4-dependent SOX2 nuclear import, and a RhoC/BCL6/FZD7 ferroptosis/Wnt branch.\",\n      \"evidence\": \"Co-IP, ubiquitination assays, nuclear fractionation, ChIP, dual luciferase, and ferroptosis/lipid-peroxidation assays across gastric and ovarian cancer cells\",\n      \"pmids\": [\"37147733\", \"37923100\", \"37060074\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether these parallel axes operate simultaneously or are context-specific is unresolved\", \"Direct ubiquitination chain-type/specificity uncharacterized\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstrated a non-cancer, immunological role by showing RNF180 degrades the m6A demethylase ALKBH5, raising SMARCA5 and worsening colitis with Th17/Treg imbalance.\",\n      \"evidence\": \"DSS-induced ulcerative colitis mouse model with knockdown/overexpression, flow cytometry, cytokine measurements, and ubiquitination analysis\",\n      \"pmids\": [\"39060657\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the same RNF180 activity is pro- or anti-disease depends on context and is unresolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Provided in vivo genetic support and broadened substrate scope into metabolism and Akt signaling, with conditional knockout enhancing colorectal carcinogenesis and degradation of NOTCH1/TRIM24/FOXC1, ACC1, and ACLY linking RNF180 to lipid metabolism and chemoresistance.\",\n      \"evidence\": \"Co-IP, ubiquitination/degradation assays, lipid droplet assays, Akt pathway analysis, and Lgr5-cre/RNF180 f/f conditional knockout mouse\",\n      \"pmids\": [\"40915184\", \"40331188\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ACC1/ACLY findings (idx 11) rest on limited methodological detail\", \"Whether all listed substrates are direct in vitro substrates not uniformly established\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Added PLK2 as a substrate linking RNF180 to EGFR/PI3K/AKT signaling and cisplatin resistance in lung cancer.\",\n      \"evidence\": \"Bioinformatic substrate prediction with qRT-PCR, Western blot, lentiviral overexpression, PLK2 knockdown, and nude mouse xenograft\",\n      \"pmids\": [\"41807497\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Substrate identified largely by bioinformatics with partial validation\", \"Direct ubiquitination not rigorously demonstrated\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how RNF180 achieves specificity across its very broad reported substrate set and which substrates are physiologically dominant in each tissue context.\",\n      \"evidence\": \"No reconstituted system defining recognition determinants or ubiquitin chain topology across substrates is present in the corpus\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural basis for substrate selection\", \"Degron/recognition motifs unmapped\", \"Chain-type specificity of ubiquitination unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 3, 4, 6]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005635\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"UbcH6\", \"RhoC\", \"DNMT1\", \"DNMT3A\", \"WISP1\", \"IPO4\", \"RAD51\", \"ALKBH5\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":7,"faith_pct":100.0}}