{"gene":"RNF115","run_date":"2026-04-28T19:45:45","timeline":{"discoveries":[{"year":2003,"finding":"Rabring7 (RNF115) specifically binds the GTP-bound form of Rab7 at its N-terminal portion, as shown by GST pull-down and co-immunoprecipitation assays. Upon binding, it is recruited from the cytosol to late endosomes/lysosomes. Overexpression causes perinuclear lysosome aggregation and impairs EGF degradation.","method":"GST pull-down, co-immunoprecipitation, live-cell imaging/LysoTracker, EGF degradation assay","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 — reciprocal pulldown + co-IP + functional localization + cellular phenotype in single foundational paper","pmids":["12972561"],"is_preprint":false},{"year":2007,"finding":"Rabring7 (RNF115) possesses E3 ubiquitin ligase activity, preferentially working with Ubc4/Ubc5 as E2 enzymes. It auto-ubiquitinates but does not ubiquitinate Rab7. A Cys229Ser mutation in the RING domain abolishes E3 ligase activity. Rabring7 accelerates EGFR degradation, while the RING mutant inhibits cCbl-induced EGFR degradation.","method":"In vitro ubiquitination assay with recombinant proteins, site-directed mutagenesis, EGFR degradation assay","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstituted E3 ligase assay with active-site mutagenesis and functional cellular validation","pmids":["17462600"],"is_preprint":false},{"year":2008,"finding":"BCA2 (RNF115) autoubiquitinates itself through its RING-H2 domain; the RING-H2 mutant lacks this activity. The BCA2 zinc finger (BZF) domain binds ubiquitin and ubiquitinated proteins. K26 and K32 mutations in the BZF domain abrogate autoubiquitination, while K232 and K260 mutations near the RING domain increase it. BCA2 mutants show altered cell migration compared to wild-type.","method":"Yeast two-hybrid/bacterial two-hybrid, in vivo ubiquitination assay, deletion and point mutagenesis, cell migration assay","journal":"Molecular cancer research","confidence":"High","confidence_rationale":"Tier 1–2 — mutagenesis of catalytic and substrate-binding residues with functional readout","pmids":["18819927"],"is_preprint":false},{"year":2009,"finding":"BCA2 (RNF115) interacts with tetherin/BST-2 and promotes tetherin-dependent restriction of HIV-1 particle production. BCA2 facilitates internalization of HIV-1 virions into CD63+ intracellular vesicles leading to lysosomal degradation. RNAi depletion of endogenous BCA2 reduces intracellular viral particle accumulation.","method":"Co-immunoprecipitation, RNAi knockdown, viral particle release assay, immunofluorescence/confocal microscopy","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-IP, RNAi loss-of-function with defined cellular phenotype, replicated by multiple experimental approaches","pmids":["20019814"],"is_preprint":false},{"year":2012,"finding":"Rabring7 (RNF115) binds MM-1, mono-ubiquitinates MM-1 in the cytoplasm without degradation, also binds c-Myc and ubiquitinates c-Myc in a Thr58-dependent manner. When co-expressed with MM-1, Rabring7 promotes c-Myc degradation. Rabring7 translocates from the cytoplasm to the nucleus together with MM-1 and c-Myc.","method":"Co-immunoprecipitation, in vivo ubiquitination assay, co-transfection/overexpression, siRNA knockdown, immunofluorescence","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP and in-cell ubiquitination with functional rescue, single lab","pmids":["22844532"],"is_preprint":false},{"year":2012,"finding":"hHR23a and 14-3-3sigma are BCA2 (RNF115) binding partners identified by yeast/bacterial two-hybrid and confirmed by in vitro binding assays. hHR23a binding dramatically reduces BCA2 autoubiquitination activity and stabilizes BCA2. Phosphorylated BCA2 is stabilized by 14-3-3sigma interaction through a pathway distinct from the proteasome.","method":"Yeast two-hybrid, bacterial two-hybrid, in vitro binding, in vivo ubiquitination assay, MG-132 proteasome inhibition","journal":"BMC cancer","confidence":"Medium","confidence_rationale":"Tier 2–3 — two-hybrid plus in vitro confirmation, single lab, multiple methods","pmids":["22315970"],"is_preprint":false},{"year":2013,"finding":"Rabring7 (RNF115) associates with EGFR through its ubiquitin-binding zinc finger domain and promotes EGFR ubiquitylation and lysosomal degradation. Rabring7 functions downstream of c-Cbl. Depletion of Rabring7 retains EGFR in a late endocytic compartment, impairs multivesicular body formation, and reduces ESCRT-II stability. Rabring7 also regulates sorting of MET and CXCR4.","method":"Co-immunoprecipitation, siRNA knockdown, in vitro ubiquitination, EGFR degradation assay, electron microscopy (MVB counting)","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 — multiple substrates, epistasis with c-Cbl, multiple orthogonal methods","pmids":["23418353"],"is_preprint":false},{"year":2013,"finding":"BCA2 (RNF115) interacts with the SUMO-conjugating enzyme UBC9 and functions as an E3 SUMO ligase, co-localizing with estrogen receptor (ER) in breast cancer cells.","method":"Co-localization/immunofluorescence, co-immunoprecipitation with UBC9, bioinformatic promoter analysis","journal":"The open cancer journal","confidence":"Low","confidence_rationale":"Tier 3 — co-localization and single co-IP; limited mechanistic follow-up for SUMO ligase activity","pmids":["21179390"],"is_preprint":false},{"year":2013,"finding":"BCA2 (RNF115) directly interacts with p21Waf1/Cip1 and promotes its K48-linked ubiquitination and proteasomal degradation, thereby promoting breast cancer cell proliferation. Knockdown of p21 partially rescues the growth arrest induced by BCA2 siRNA.","method":"Co-immunoprecipitation, in vivo ubiquitination assay, siRNA knockdown, cell proliferation assay, genetic epistasis (siRNA rescue)","journal":"Neoplasia","confidence":"High","confidence_rationale":"Tier 2 — direct interaction, in-cell ubiquitination, genetic epistasis rescue, multiple breast cancer cell lines","pmids":["24027428"],"is_preprint":false},{"year":2017,"finding":"BCA2 (RNF115) acts as an E3 SUMO ligase for IκBα, enhancing IκBα SUMOylation, which sequesters NF-κB in the cytoplasm and reduces NF-κB-dependent HIV proviral transcription by up to 4.4-fold in CD4+ T cells. BCA2 is induced by NF-κB-activating cytokines and provides negative feedback on this pathway.","method":"In-cell SUMOylation assay, NF-κB reporter assay, siRNA knockdown, HIV transcription measurement in CD4+ T cells","journal":"Journal of virology","confidence":"Medium","confidence_rationale":"Tier 2 — functional SUMO ligase activity demonstrated in cells with mechanistic pathway placement, single lab","pmids":["28122985"],"is_preprint":false},{"year":2020,"finding":"RNF115 constitutively interacts with MAVS and catalyzes K48-linked ubiquitination and proteasomal degradation of MAVS in uninfected cells, maintaining basal MAVS levels. After HSV-1 infection, RNF115 associates with MITA/STING and catalyzes K63-linked ubiquitination to promote MITA aggregation and antiviral signaling. Rnf115-/- mice show elevated MAVS protein basally and impaired HSV-1-induced MITA aggregation.","method":"Co-immunoprecipitation, in vivo ubiquitination assay (K48 and K63 linkage-specific), Rnf115 knockout mice, MITA aggregation assay, viral infection models","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 — dual substrate characterization with linkage-specific ubiquitination, in vivo knockout model with multiple phenotypic readouts, replicated across conditions","pmids":["33139700"],"is_preprint":false},{"year":2020,"finding":"RNF115 interacts with STX17 (syntaxin-17) and enhances its stability, which is required for autophagosome-lysosome fusion. RNF115 deletion causes accumulation of autophagosomes by impairing their fusion with lysosomes.","method":"Co-immunoprecipitation, autophagosome/lysosome fusion assay, RNF115 knockout cells, LC3/STX17 protein stability assay","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2–3 — co-IP with functional consequence in KO cells, single lab","pmids":["32980859"],"is_preprint":false},{"year":2020,"finding":"RNF115 mediates ubiquitination of p53, promoting its degradation in lung adenocarcinoma cells. RNF115 disruption induces G1 phase arrest and inhibits tumor growth in xenograft models.","method":"In vivo ubiquitination assay, siRNA/shRNA knockdown, cell cycle analysis, xenograft mouse model","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2–3 — ubiquitination assay plus in vivo functional data, single lab","pmids":["32553631"],"is_preprint":false},{"year":2021,"finding":"RNF115 promotes Wnt/β-catenin pathway activation by mediating ubiquitination and proteasomal degradation of APC (adenomatous polyposis coli) in lung adenocarcinoma cells.","method":"Co-immunoprecipitation, ubiquitination assay, RNF115 knockdown/overexpression, β-catenin pathway reporter","journal":"Cancer & metabolism","confidence":"Medium","confidence_rationale":"Tier 2–3 — ubiquitination of APC shown with functional Wnt pathway readout, single lab","pmids":["33509267"],"is_preprint":false},{"year":2022,"finding":"RNF115 is phosphorylated by AKT1, enabling 14-3-3 chaperones to bind phosphorylated RNF115 and facilitate its localization to the ER and Golgi apparatus. RNF115 then interacts with RAB1A and RAB13 and catalyzes K11-linked ubiquitination on specific lysine residues (Lys49/Lys61 of RAB1A; Lys46/Lys58 of RAB13), impairing GDI1 recruitment to RAB proteins and thereby blocking post-ER trafficking of Toll-like receptors.","method":"Co-immunoprecipitation, in vivo ubiquitination assay (K11-specific), site-directed mutagenesis, Rnf115 KO cells, subcellular fractionation/localization, TLR trafficking assay, AKT1 kinase assay","journal":"Advanced science","confidence":"High","confidence_rationale":"Tier 1–2 — linkage-specific ubiquitination with defined sites via mutagenesis, upstream kinase identified, downstream GDI1 mechanism, KO rescue experiments","pmids":["35343654"],"is_preprint":false},{"year":2022,"finding":"RNF115 is enriched on phagosomes of IFN-γ-activated macrophages and acts as a regulator of phagosomal maturation. Loss of RNF115 protein or its ligase activity enhances phagosomal maturation and increases cytokine responses to bacterial infection. RNF115 KO mice show less tissue damage in response to S. aureus infection.","method":"Quantitative proteomics of phagosomes, RNF115 KO (cells and mice), phagosomal maturation assay, bacterial infection assay, cytokine measurement","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — phagosomal proteomics + genetic KO with in vivo validation, multiple orthogonal methods","pmids":["36281581"],"is_preprint":false},{"year":2023,"finding":"RNF115 interacts with LC3B and downregulates LC3B protein levels, suppressing autophagy. RNF115 deletion promotes autophagy, reduces damaged mitochondria, and protects against LPS/D-GalN-induced acute liver injury in mice via NF-κB and JNK pathway inhibition.","method":"Co-immunoprecipitation (RNF115-LC3B), RNF115 KO mice, autophagy flux assay, adeno-associated virus rescue, pharmacological autophagy inhibition (3-MA), cytokine/macrophage analysis","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2–3 — co-IP with LC3B plus in vivo KO, but mechanistic detail of LC3B regulation is limited","pmids":["38129372"],"is_preprint":false},{"year":2024,"finding":"RNF115 ubiquitinates and promotes proteasomal degradation of CDK10, activating the Raf-1 pathway and promoting cell cycle progression in thyroid carcinoma cells. CDK10 overexpression rescues RNF115-induced malignant phenotypes.","method":"Co-immunoprecipitation, ubiquitination assay, RNF115 KO/overexpression, CDK10 overexpression rescue, Raf-1 pathway analysis","journal":"Cell biology and toxicology","confidence":"Medium","confidence_rationale":"Tier 2–3 — ubiquitination of CDK10 with pathway placement by rescue experiment, single lab","pmids":["38376606"],"is_preprint":false},{"year":2024,"finding":"Knockout or pharmacological inhibition of RNF115 (with disulfiram) impairs STING oligomerization and Golgi localization, thereby inhibiting STING-mediated inflammatory signaling. This effect is cell-type specific: RNF115 knockout inhibits STING activation in myeloid cells but not in endothelial cells or fibroblasts.","method":"RNF115 KO mice, bone marrow chimeric mice, disulfiram pharmacological inhibition, STING oligomerization assay, Golgi localization assay, PBMC cytokine measurement","journal":"Cellular & molecular immunology","confidence":"High","confidence_rationale":"Tier 2 — in vivo KO and pharmacological inhibition with mechanistic readout across multiple disease models","pmids":["38267694"],"is_preprint":false},{"year":2025,"finding":"RNF115 acts as an E3 ubiquitin ligase for DHODH, promoting K27-linked ubiquitination of DHODH and inhibiting its autophagic degradation, thereby suppressing ferroptosis in hepatocellular carcinoma. YBX1 promotes RNF115 mRNA translation through m5C modification of the 3'-UTR.","method":"Co-immunoprecipitation, ubiquitination assay (K27-linkage), autophagic degradation assay, RNF115 KO/overexpression, RIP-seq, MeRIP-qPCR, dual-luciferase reporter","journal":"Clinical and translational medicine","confidence":"Medium","confidence_rationale":"Tier 2–3 — K27-ubiquitination of DHODH with functional ferroptosis readout; translational regulation by m5C is well-supported but single lab","pmids":["40088428"],"is_preprint":false},{"year":2026,"finding":"RNF115 coordinates with the adaptor Tollip to mediate selective autophagic degradation of PEDV nucleocapsid (N) protein via K63-linked ubiquitination at residues K271, K276, K302, K359, and K375. The C25 and C38 residues of RNF115 are critical for N protein ubiquitination. Tollip forms a stable complex with RNF115 and N protein to facilitate substrate recognition.","method":"Co-immunoprecipitation, in vivo ubiquitination assay (K63-specific), site-directed mutagenesis, siRNA knockdown of Tollip, autophagic flux assay, viral replication assay","journal":"International journal of biological macromolecules","confidence":"Medium","confidence_rationale":"Tier 2 — site-specific ubiquitination with mutagenesis and adaptor identification, single lab, recent paper","pmids":["41587704"],"is_preprint":false}],"current_model":"RNF115 (BCA2/Rabring7) is a RING-H2-type E3 ubiquitin ligase (and E3 SUMO ligase) that uses its N-terminal BZF domain to bind ubiquitin/ubiquitinated proteins and its RING domain to catalyze autoubiquitination as well as K11-, K27-, K48-, and K63-linked ubiquitination of diverse substrates—including MAVS, MITA/STING, RAB1A, RAB13, p21, p53, APC, CDK10, EGFR, c-Myc, LC3B, and DHODH—thereby regulating innate antiviral and antibacterial immune responses, endolysosomal trafficking (via Rab7 interaction and tetherin cofactor activity), phagosomal maturation, autophagy, and cancer cell proliferation; its activity and localization are modulated by AKT1 phosphorylation and 14-3-3 binding, and by partner proteins such as hHR23a."},"narrative":{"teleology":[{"year":2003,"claim":"The discovery that RNF115 binds GTP-Rab7 and is recruited to late endosomes/lysosomes established it as a regulator of endolysosomal trafficking, explaining why its overexpression causes perinuclear lysosome aggregation and impairs EGF degradation.","evidence":"GST pull-down, co-immunoprecipitation, LysoTracker imaging, and EGF degradation assay in mammalian cells","pmids":["12972561"],"confidence":"High","gaps":["Rab7 was not shown to be a direct ubiquitination substrate","mechanism of lysosome aggregation was unclear","physiological stoichiometry of the interaction was not determined"]},{"year":2007,"claim":"Reconstitution of E3 ubiquitin ligase activity with Ubc4/Ubc5 E2 enzymes and identification of the catalytic Cys229 residue in the RING domain established RNF115 as a bona fide ubiquitin ligase that accelerates EGFR degradation.","evidence":"In vitro ubiquitination assay with recombinant proteins, C229S mutagenesis, EGFR degradation assay","pmids":["17462600"],"confidence":"High","gaps":["Direct EGFR ubiquitination by RNF115 was not shown","ubiquitin chain linkage type was not determined","relationship to c-Cbl was not resolved"]},{"year":2008,"claim":"Mapping of the BZF domain as a ubiquitin-binding module and identification of autoubiquitination-regulatory lysines revealed a bipartite architecture in which substrate recognition (BZF) and catalysis (RING-H2) are separable functions.","evidence":"Yeast/bacterial two-hybrid, mutagenesis of K26/K32 (BZF) and K232/K260 (RING-proximal), in vivo ubiquitination assay","pmids":["18819927"],"confidence":"High","gaps":["Crystal structure of the BZF–ubiquitin complex was not determined","how BZF-mediated ubiquitin binding directs substrate selectivity was unknown"]},{"year":2009,"claim":"Demonstration that RNF115 cooperates with tetherin/BST-2 to internalize HIV-1 virions into CD63+ endosomes for lysosomal degradation revealed a direct antiviral effector function beyond general endosomal trafficking.","evidence":"Co-immunoprecipitation with tetherin, RNAi knockdown, viral particle release assay, confocal microscopy in HeLa cells","pmids":["20019814"],"confidence":"High","gaps":["Whether RNF115 ubiquitinates tetherin or viral proteins was not determined","mechanism of virion internalization was unclear"]},{"year":2012,"claim":"Identification of hHR23a and 14-3-3σ as binding partners that stabilize RNF115 by suppressing autoubiquitination and by phosphorylation-dependent sequestration, respectively, revealed that RNF115 turnover is tightly regulated by opposing stabilization mechanisms.","evidence":"Yeast/bacterial two-hybrid, in vitro binding, in vivo ubiquitination assay with proteasome inhibition","pmids":["22315970"],"confidence":"Medium","gaps":["The kinase responsible for RNF115 phosphorylation was not identified in this study","in vivo relevance was not tested"]},{"year":2013,"claim":"Establishing that RNF115 functions downstream of c-Cbl to ubiquitinate EGFR and promote multivesicular body sorting—and extends to MET and CXCR4—positioned it as a general regulator of receptor tyrosine kinase and GPCR endosomal sorting via ESCRT-II stabilization.","evidence":"Co-immunoprecipitation, siRNA knockdown, in vitro ubiquitination, electron microscopy of MVBs in HeLa cells","pmids":["23418353"],"confidence":"High","gaps":["Direct ubiquitination of ESCRT-II components was not tested","structural basis for broad receptor substrate recognition was unknown"]},{"year":2013,"claim":"Discovery that RNF115 ubiquitinates p21Waf1/Cip1 via K48 linkage for proteasomal degradation, with genetic epistasis rescue, provided the first mechanistic link between RNF115 E3 activity and cell cycle control in breast cancer.","evidence":"Co-immunoprecipitation, K48-linkage-specific ubiquitination, p21 siRNA rescue of BCA2 knockdown growth arrest in breast cancer cell lines","pmids":["24027428"],"confidence":"High","gaps":["Whether RNF115-mediated p21 degradation occurs in non-transformed cells was unknown","contribution relative to other p21 E3 ligases was not assessed"]},{"year":2017,"claim":"Demonstration that RNF115 SUMOylates IκBα to sequester NF-κB in the cytoplasm established a dual E3 ligase function (ubiquitin and SUMO) and identified a negative feedback loop restraining NF-κB-driven HIV transcription in CD4+ T cells.","evidence":"In-cell SUMOylation assay, NF-κB reporter, siRNA knockdown, HIV transcription measurement in CD4+ T cells","pmids":["28122985"],"confidence":"Medium","gaps":["SUMO ligase activity was not reconstituted in vitro with purified components","whether this mechanism operates beyond HIV infection was untested"]},{"year":2020,"claim":"Using Rnf115 knockout mice, the dual and context-dependent role of RNF115 in innate immunity was established: constitutive K48-linked ubiquitination of MAVS maintains basal signaling tone, while infection-induced K63-linked ubiquitination of STING/MITA drives antiviral defense.","evidence":"Co-immunoprecipitation, linkage-specific ubiquitination assays, Rnf115−/− mice, MITA aggregation assay, viral infection models","pmids":["33139700"],"confidence":"High","gaps":["Signal that triggers the MAVS-to-STING substrate switch was not identified","whether other E3 ligases compensate in knockout animals was not addressed"]},{"year":2020,"claim":"The finding that RNF115 stabilizes STX17 to promote autophagosome–lysosome fusion, and that its loss causes autophagosome accumulation, extended RNF115's role from endosomal trafficking to autophagy regulation.","evidence":"Co-immunoprecipitation with STX17, RNF115 KO cells, autophagosome-lysosome fusion assay","pmids":["32980859"],"confidence":"Medium","gaps":["Whether STX17 is a direct ubiquitination substrate was not shown","ubiquitin linkage type on STX17 was not determined"]},{"year":2022,"claim":"Identification of AKT1 as the upstream kinase that phosphorylates RNF115 to enable 14-3-3 binding and ER/Golgi relocalization, where RNF115 catalyzes K11-linked ubiquitination of RAB1A/RAB13 to block TLR post-ER trafficking, provided a complete signaling-to-substrate mechanism for immune regulation.","evidence":"AKT1 kinase assay, K11-linkage-specific ubiquitination, site-directed mutagenesis of RAB lysines, Rnf115 KO cells, subcellular fractionation, TLR trafficking assay","pmids":["35343654"],"confidence":"High","gaps":["Whether AKT1-mediated regulation applies to all RNF115 substrates was unknown","structural basis for K11-linkage selectivity was not determined"]},{"year":2022,"claim":"Phagosomal proteomics in IFN-γ-activated macrophages revealed RNF115 enrichment on phagosomes, with knockout showing enhanced phagosomal maturation and reduced tissue damage in S. aureus infection, establishing RNF115 as a brake on antimicrobial phagosome function.","evidence":"Quantitative phagosomal proteomics, RNF115 KO cells and mice, phagosomal maturation assay, S. aureus infection","pmids":["36281581"],"confidence":"High","gaps":["The phagosomal substrate(s) of RNF115 ligase activity were not identified","mechanism distinguishing phagosomal from endosomal function was unknown"]},{"year":2023,"claim":"RNF115 was shown to downregulate LC3B protein levels to suppress autophagy, and its deletion protected against LPS/D-GalN-induced acute liver injury in mice through reduced NF-κB and JNK activation, linking its autophagy-regulatory role to organ-level inflammatory damage.","evidence":"Co-immunoprecipitation with LC3B, RNF115 KO mice, autophagy flux assay, AAV rescue, 3-MA inhibition","pmids":["38129372"],"confidence":"Medium","gaps":["Whether LC3B is directly ubiquitinated by RNF115 and via which linkage was not defined","relative contribution of autophagy versus NF-κB suppression to the protective phenotype was unclear"]},{"year":2024,"claim":"Demonstration that RNF115 knockout or pharmacological inhibition with disulfiram impairs STING oligomerization and Golgi localization specifically in myeloid cells refined the cell-type-specific model of RNF115-dependent innate immune control and suggested therapeutic tractability.","evidence":"RNF115 KO mice, bone marrow chimeras, disulfiram treatment, STING oligomerization and Golgi localization assays, PBMC cytokine measurements","pmids":["38267694"],"confidence":"High","gaps":["Direct target of disulfiram inhibition on RNF115 catalytic mechanism was not resolved","basis for myeloid-cell-type specificity was unknown"]},{"year":2025,"claim":"Identification of DHODH as a K27-linked ubiquitination substrate whose modification by RNF115 inhibits autophagic degradation and suppresses ferroptosis revealed a new linkage type and an unexpected role in metabolic cell death regulation in hepatocellular carcinoma.","evidence":"K27-linkage-specific ubiquitination assay, autophagic degradation assay, RNF115 KO/overexpression, RIP-seq and MeRIP-qPCR for m5C-dependent translational regulation","pmids":["40088428"],"confidence":"Medium","gaps":["K27-ubiquitination mechanism was not reconstituted in vitro","structural basis for linkage selectivity among K11, K27, K48, K63 across substrates remains unresolved"]},{"year":null,"claim":"How RNF115 selects among K11, K27, K48, and K63 ubiquitin linkage types for different substrates, and what determines the context-dependent switch between constitutive degradative versus signal-amplifying ubiquitination programs, remain central unresolved questions.","evidence":"","pmids":[],"confidence":"High","gaps":["No structural model of full-length RNF115 or its substrate complexes exists","E2 enzyme pairing rules that dictate linkage specificity are unknown","how AKT1 phosphorylation and 14-3-3 binding integrate with substrate-specific switches has not been resolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[1,2,6,8,10,12,13,14,17,19,20]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[1,8,10,14,19]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[2]}],"localization":[{"term_id":"GO:0005764","term_label":"lysosome","supporting_discovery_ids":[0,3,6]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,4]},{"term_id":"GO:0005783","term_label":"endoplasmic reticulum","supporting_discovery_ids":[14]},{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[14]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[4]},{"term_id":"GO:0005768","term_label":"endosome","supporting_discovery_ids":[0,6]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[10,14,15,18]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[1,6,8,10,14]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[0,3,6,14]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[11,16,20]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[8,12,17]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[9,13,14]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[19]}],"complexes":[],"partners":["RAB7A","MAVS","TMEM173","BST2","RAB1A","RAB13","STX17","TOLLIP"],"other_free_text":[]},"mechanistic_narrative":"RNF115 (BCA2/Rabring7) is a RING-H2-type E3 ubiquitin ligase that controls endolysosomal trafficking, innate immune signaling, autophagy, and cell proliferation by catalyzing K11-, K27-, K48-, and K63-linked ubiquitination of diverse substrates. Its N-terminal BCA2 zinc finger (BZF) domain binds ubiquitin and ubiquitinated proteins to recruit substrates, while the RING-H2 domain mediates catalytic activity with Ubc4/Ubc5 family E2 enzymes; AKT1 phosphorylation and 14-3-3 binding regulate its ER/Golgi localization and activity [PMID:18819927, PMID:17462600, PMID:35343654]. In innate immunity, RNF115 constitutively targets MAVS for K48-linked degradation to set basal levels, and upon viral infection switches to K63-linked ubiquitination of STING/MITA to promote its oligomerization and antiviral signaling, while on phagosomes it restrains phagosomal maturation during bacterial infection [PMID:33139700, PMID:38267694, PMID:36281581]. RNF115 also promotes cancer cell proliferation by ubiquitinating and degrading tumor suppressors p21 and p53, the Wnt pathway component APC, and CDK10, and regulates autophagy through interactions with STX17 and LC3B [PMID:24027428, PMID:32553631, PMID:33509267, PMID:38376606, PMID:32980859, PMID:38129372]."},"prefetch_data":{"uniprot":{"accession":"Q9Y4L5","full_name":"E3 ubiquitin-protein ligase RNF115","aliases":["RING finger protein 115","RING-type E3 ubiquitin transferase RNF115","Rab7-interacting RING finger protein","Rabring 7","Zinc finger protein 364"],"length_aa":304,"mass_kda":33.7,"function":"E3 ubiquitin-protein ligase that catalyzes the 'Lys-48'- and/or 'Lys-63'-linked polyubiquitination of various substrates and thereby plays a role in a number of signaling pathways including autophagy, innate immunity, cell proliferation and cell death (PubMed:20019814, PubMed:30689267). Plays a role in the endosomal trafficking and degradation of membrane receptors including EGFR, FLT3, MET and CXCR4 through their polyubiquitination. Participates together with BST2 in antiviral immunity by facilitating the internalization of HIV-1 virions into intracellular vesicles leading to their lysosomal degradation (PubMed:20019814). Also possesses an antiviral activity independently of BST2 by promoting retroviral GAG proteins ubiquitination, redistribution to endo-lysosomal compartments and, ultimately, lysosomal degradation (PubMed:24852021). Catalyzes distinct types of ubiquitination on MAVS and STING1 at different phases of viral infection to promote innate antiviral response (PubMed:33139700). Mediates the 'Lys-48'-linked ubiquitination of MAVS leading to its proteasomal degradation and ubiquitinates STING1 via 'Lys-63'-linked polyubiquitination, critical for its oligomerization and the subsequent recruitment of TBK1 (PubMed:33139700). Plays a positive role in the autophagosome-lysosome fusion by interacting with STX17 and enhancing its stability without affecting 'Lys-48'- or 'Lys-63'-linked polyubiquitination levels, which in turn promotes autophagosome maturation (PubMed:32980859). Negatively regulates TLR-induced expression of proinflammatory cytokines by catalyzing 'Lys-11'-linked ubiquitination of RAB1A and RAB13 to inhibit post-ER trafficking of TLRs to the Golgi by RAB1A and subsequently from the Golgi apparatus to the cell surface by RAB13 (PubMed:35343654)","subcellular_location":"Cytoplasm; Nucleus; Endoplasmic reticulum; Golgi apparatus","url":"https://www.uniprot.org/uniprotkb/Q9Y4L5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF115","classification":"Not Classified","n_dependent_lines":30,"n_total_lines":1208,"dependency_fraction":0.024834437086092714},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF115","total_profiled":1310},"omim":[{"mim_id":"619535","title":"RING FINGER PROTEIN 115; RNF115","url":"https://www.omim.org/entry/619535"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoli","reliability":"Approved"},{"location":"Mitochondria","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RNF115"},"hgnc":{"alias_symbol":["BCA2","Rabring7","CL469780"],"prev_symbol":["ZNF364"]},"alphafold":{"accession":"Q9Y4L5","domains":[{"cath_id":"-","chopping":"20-50","consensus_level":"high","plddt":85.2019,"start":20,"end":50},{"cath_id":"3.30.40.10","chopping":"207-262","consensus_level":"high","plddt":82.9345,"start":207,"end":262}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y4L5","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y4L5-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y4L5-F1-predicted_aligned_error_v6.png","plddt_mean":59.84},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF115","jax_strain_url":"https://www.jax.org/strain/search?query=RNF115"},"sequence":{"accession":"Q9Y4L5","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y4L5.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y4L5/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y4L5"}},"corpus_meta":[{"pmid":"20222671","id":"PMC_20222671","title":"Exploring the structural requirements for inhibition of the ubiquitin E3 ligase breast cancer associated protein 2 (BCA2) as a treatment for breast cancer.","date":"2010","source":"Journal of medicinal chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/20222671","citation_count":98,"is_preprint":false},{"pmid":"33139700","id":"PMC_33139700","title":"RNF115 plays dual roles in innate antiviral responses by catalyzing distinct ubiquitination of MAVS and MITA.","date":"2020","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/33139700","citation_count":78,"is_preprint":false},{"pmid":"20019814","id":"PMC_20019814","title":"BCA2/Rabring7 promotes tetherin-dependent HIV-1 restriction.","date":"2009","source":"PLoS pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/20019814","citation_count":78,"is_preprint":false},{"pmid":"12972561","id":"PMC_12972561","title":"Rabring7, a novel Rab7 target protein with a RING finger motif.","date":"2003","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/12972561","citation_count":76,"is_preprint":false},{"pmid":"18819927","id":"PMC_18819927","title":"Autoubiquitination of BCA2 RING E3 ligase regulates its own stability and affects cell migration.","date":"2008","source":"Molecular cancer research : MCR","url":"https://pubmed.ncbi.nlm.nih.gov/18819927","citation_count":70,"is_preprint":false},{"pmid":"23418353","id":"PMC_23418353","title":"The E3 ubiquitin ligases RNF126 and Rabring7 regulate endosomal sorting of the epidermal growth factor receptor.","date":"2013","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/23418353","citation_count":48,"is_preprint":false},{"pmid":"17462600","id":"PMC_17462600","title":"Involvement of Rabring7 in EGF receptor degradation as an E3 ligase.","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/17462600","citation_count":33,"is_preprint":false},{"pmid":"35343654","id":"PMC_35343654","title":"RNF115 Inhibits the Post-ER Trafficking of TLRs and TLRs-Mediated Immune Responses by Catalyzing K11-Linked Ubiquitination of RAB1A and RAB13.","date":"2022","source":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","url":"https://pubmed.ncbi.nlm.nih.gov/35343654","citation_count":32,"is_preprint":false},{"pmid":"33509267","id":"PMC_33509267","title":"RNF115 promotes lung adenocarcinoma through Wnt/β-catenin pathway activation by mediating APC ubiquitination.","date":"2021","source":"Cancer & metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/33509267","citation_count":29,"is_preprint":false},{"pmid":"24027428","id":"PMC_24027428","title":"RNF115/BCA2 E3 ubiquitin ligase promotes breast cancer cell proliferation through targeting p21Waf1/Cip1 for ubiquitin-mediated degradation.","date":"2013","source":"Neoplasia (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/24027428","citation_count":26,"is_preprint":false},{"pmid":"22844532","id":"PMC_22844532","title":"Rabring7 degrades c-Myc through complex formation with MM-1.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/22844532","citation_count":24,"is_preprint":false},{"pmid":"28122985","id":"PMC_28122985","title":"BCA2/Rabring7 Interferes with HIV-1 Proviral Transcription by Enhancing the SUMOylation of IκBα.","date":"2017","source":"Journal of virology","url":"https://pubmed.ncbi.nlm.nih.gov/28122985","citation_count":23,"is_preprint":false},{"pmid":"38267694","id":"PMC_38267694","title":"Disulfiram ameliorates STING/MITA-dependent inflammation and autoimmunity by targeting RNF115.","date":"2024","source":"Cellular & molecular immunology","url":"https://pubmed.ncbi.nlm.nih.gov/38267694","citation_count":22,"is_preprint":false},{"pmid":"22315970","id":"PMC_22315970","title":"Effects of partner proteins on BCA2 RING ligase activity.","date":"2012","source":"BMC cancer","url":"https://pubmed.ncbi.nlm.nih.gov/22315970","citation_count":22,"is_preprint":false},{"pmid":"36281581","id":"PMC_36281581","title":"The E3 ubiquitin ligase RNF115 regulates phagosome maturation and host response to bacterial infection.","date":"2022","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/36281581","citation_count":20,"is_preprint":false},{"pmid":"32147403","id":"PMC_32147403","title":"Roles of RNF126 and BCA2 E3 ubiquitin ligases in DNA damage repair signaling and targeted cancer therapy.","date":"2020","source":"Pharmacological research","url":"https://pubmed.ncbi.nlm.nih.gov/32147403","citation_count":20,"is_preprint":false},{"pmid":"40088428","id":"PMC_40088428","title":"Targeting YBX1-m5C mediates RNF115 mRNA circularisation and translation to enhance vulnerability of ferroptosis in hepatocellular carcinoma.","date":"2025","source":"Clinical and translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/40088428","citation_count":19,"is_preprint":false},{"pmid":"38129372","id":"PMC_38129372","title":"RNF115/BCA2 deficiency alleviated acute liver injury in mice by promoting autophagy and inhibiting inflammatory response.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/38129372","citation_count":18,"is_preprint":false},{"pmid":"21179390","id":"PMC_21179390","title":"Role of the BCA2 ubiquitin E3 ligase in hormone responsive breast cancer.","date":"2010","source":"The open cancer journal","url":"https://pubmed.ncbi.nlm.nih.gov/21179390","citation_count":18,"is_preprint":false},{"pmid":"32553631","id":"PMC_32553631","title":"RNF115-mediated ubiquitination of p53 regulates lung adenocarcinoma proliferation.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/32553631","citation_count":17,"is_preprint":false},{"pmid":"32980859","id":"PMC_32980859","title":"RNF115 deletion inhibits autophagosome maturation and growth of gastric cancer.","date":"2020","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/32980859","citation_count":16,"is_preprint":false},{"pmid":"23951330","id":"PMC_23951330","title":"Expansion and diversification of BTL ring-H2 ubiquitin ligases in angiosperms: putative Rabring7/BCA2 orthologs.","date":"2013","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23951330","citation_count":13,"is_preprint":false},{"pmid":"23242606","id":"PMC_23242606","title":"BCA2 is differentially expressed in renal oncocytoma: an analysis of 158 renal neoplasms.","date":"2012","source":"Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine","url":"https://pubmed.ncbi.nlm.nih.gov/23242606","citation_count":10,"is_preprint":false},{"pmid":"38376606","id":"PMC_38376606","title":"RNF115 aggravates tumor progression through regulation of CDK10 degradation in thyroid carcinoma.","date":"2024","source":"Cell biology and toxicology","url":"https://pubmed.ncbi.nlm.nih.gov/38376606","citation_count":8,"is_preprint":false},{"pmid":"34589482","id":"PMC_34589482","title":"Effects of the SUMO Ligase BCA2 on Metabolic Activity, Cell Proliferation, Cell Migration, Cell Cycle, and the Regulation of NF-κB and IRF1 in Different Breast Epithelial Cellular Contexts.","date":"2021","source":"Frontiers in cell and developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/34589482","citation_count":7,"is_preprint":false},{"pmid":"38725852","id":"PMC_38725852","title":"E3 ubiquitin ligase BCA2 promotes breast cancer stemness by up-regulation of SOX9 by LPS.","date":"2024","source":"International journal of biological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/38725852","citation_count":5,"is_preprint":false},{"pmid":"28964893","id":"PMC_28964893","title":"Autoubiquitination of feline E3 ubiquitin ligase BCA2.","date":"2017","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/28964893","citation_count":5,"is_preprint":false},{"pmid":"32595622","id":"PMC_32595622","title":"Antiviral Activity of Feline BCA2 Is Mainly Dependent on Its Interference With Proviral Transcription Rather Than Degradation of FIV Gag.","date":"2020","source":"Frontiers in microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/32595622","citation_count":5,"is_preprint":false},{"pmid":"26208753","id":"PMC_26208753","title":"Characterization and expression analysis of BCA2 gene in large yellow croaker, Larimichthys crocea.","date":"2015","source":"Fish & shellfish immunology","url":"https://pubmed.ncbi.nlm.nih.gov/26208753","citation_count":5,"is_preprint":false},{"pmid":"16473630","id":"PMC_16473630","title":"Rabring7: a target protein for rab7 small g protein.","date":"2005","source":"Methods in enzymology","url":"https://pubmed.ncbi.nlm.nih.gov/16473630","citation_count":1,"is_preprint":false},{"pmid":"41272272","id":"PMC_41272272","title":"RNF115 upregulation by YBX1-dependent m5C modification promotes hepatocellular carcinoma progression.","date":"2025","source":"NPJ precision oncology","url":"https://pubmed.ncbi.nlm.nih.gov/41272272","citation_count":1,"is_preprint":false},{"pmid":"41587704","id":"PMC_41587704","title":"RNF115-Tollip coordination mediates selective autophagic degradation of N protein during PEDV infection.","date":"2026","source":"International journal of biological macromolecules","url":"https://pubmed.ncbi.nlm.nih.gov/41587704","citation_count":0,"is_preprint":false},{"pmid":"40079889","id":"PMC_40079889","title":"Diaphorina citri E3 ubiquitin ligase RNF115 inhibits CLas bacterial proliferation by targeting to the host histone H1.","date":"2025","source":"Insect science","url":"https://pubmed.ncbi.nlm.nih.gov/40079889","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":17841,"output_tokens":5392,"usd":0.067201},"stage2":{"model":"claude-opus-4-6","input_tokens":8964,"output_tokens":4267,"usd":0.227242},"total_usd":0.294443,"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\": 2003,\n      \"finding\": \"Rabring7 (RNF115) specifically binds the GTP-bound form of Rab7 at its N-terminal portion, as shown by GST pull-down and co-immunoprecipitation assays. Upon binding, it is recruited from the cytosol to late endosomes/lysosomes. Overexpression causes perinuclear lysosome aggregation and impairs EGF degradation.\",\n      \"method\": \"GST pull-down, co-immunoprecipitation, live-cell imaging/LysoTracker, EGF degradation assay\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal pulldown + co-IP + functional localization + cellular phenotype in single foundational paper\",\n      \"pmids\": [\"12972561\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Rabring7 (RNF115) possesses E3 ubiquitin ligase activity, preferentially working with Ubc4/Ubc5 as E2 enzymes. It auto-ubiquitinates but does not ubiquitinate Rab7. A Cys229Ser mutation in the RING domain abolishes E3 ligase activity. Rabring7 accelerates EGFR degradation, while the RING mutant inhibits cCbl-induced EGFR degradation.\",\n      \"method\": \"In vitro ubiquitination assay with recombinant proteins, site-directed mutagenesis, EGFR degradation assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstituted E3 ligase assay with active-site mutagenesis and functional cellular validation\",\n      \"pmids\": [\"17462600\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"BCA2 (RNF115) autoubiquitinates itself through its RING-H2 domain; the RING-H2 mutant lacks this activity. The BCA2 zinc finger (BZF) domain binds ubiquitin and ubiquitinated proteins. K26 and K32 mutations in the BZF domain abrogate autoubiquitination, while K232 and K260 mutations near the RING domain increase it. BCA2 mutants show altered cell migration compared to wild-type.\",\n      \"method\": \"Yeast two-hybrid/bacterial two-hybrid, in vivo ubiquitination assay, deletion and point mutagenesis, cell migration assay\",\n      \"journal\": \"Molecular cancer research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — mutagenesis of catalytic and substrate-binding residues with functional readout\",\n      \"pmids\": [\"18819927\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"BCA2 (RNF115) interacts with tetherin/BST-2 and promotes tetherin-dependent restriction of HIV-1 particle production. BCA2 facilitates internalization of HIV-1 virions into CD63+ intracellular vesicles leading to lysosomal degradation. RNAi depletion of endogenous BCA2 reduces intracellular viral particle accumulation.\",\n      \"method\": \"Co-immunoprecipitation, RNAi knockdown, viral particle release assay, immunofluorescence/confocal microscopy\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP, RNAi loss-of-function with defined cellular phenotype, replicated by multiple experimental approaches\",\n      \"pmids\": [\"20019814\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Rabring7 (RNF115) binds MM-1, mono-ubiquitinates MM-1 in the cytoplasm without degradation, also binds c-Myc and ubiquitinates c-Myc in a Thr58-dependent manner. When co-expressed with MM-1, Rabring7 promotes c-Myc degradation. Rabring7 translocates from the cytoplasm to the nucleus together with MM-1 and c-Myc.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay, co-transfection/overexpression, siRNA knockdown, immunofluorescence\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP and in-cell ubiquitination with functional rescue, single lab\",\n      \"pmids\": [\"22844532\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"hHR23a and 14-3-3sigma are BCA2 (RNF115) binding partners identified by yeast/bacterial two-hybrid and confirmed by in vitro binding assays. hHR23a binding dramatically reduces BCA2 autoubiquitination activity and stabilizes BCA2. Phosphorylated BCA2 is stabilized by 14-3-3sigma interaction through a pathway distinct from the proteasome.\",\n      \"method\": \"Yeast two-hybrid, bacterial two-hybrid, in vitro binding, in vivo ubiquitination assay, MG-132 proteasome inhibition\",\n      \"journal\": \"BMC cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — two-hybrid plus in vitro confirmation, single lab, multiple methods\",\n      \"pmids\": [\"22315970\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Rabring7 (RNF115) associates with EGFR through its ubiquitin-binding zinc finger domain and promotes EGFR ubiquitylation and lysosomal degradation. Rabring7 functions downstream of c-Cbl. Depletion of Rabring7 retains EGFR in a late endocytic compartment, impairs multivesicular body formation, and reduces ESCRT-II stability. Rabring7 also regulates sorting of MET and CXCR4.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, in vitro ubiquitination, EGFR degradation assay, electron microscopy (MVB counting)\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple substrates, epistasis with c-Cbl, multiple orthogonal methods\",\n      \"pmids\": [\"23418353\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"BCA2 (RNF115) interacts with the SUMO-conjugating enzyme UBC9 and functions as an E3 SUMO ligase, co-localizing with estrogen receptor (ER) in breast cancer cells.\",\n      \"method\": \"Co-localization/immunofluorescence, co-immunoprecipitation with UBC9, bioinformatic promoter analysis\",\n      \"journal\": \"The open cancer journal\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — co-localization and single co-IP; limited mechanistic follow-up for SUMO ligase activity\",\n      \"pmids\": [\"21179390\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"BCA2 (RNF115) directly interacts with p21Waf1/Cip1 and promotes its K48-linked ubiquitination and proteasomal degradation, thereby promoting breast cancer cell proliferation. Knockdown of p21 partially rescues the growth arrest induced by BCA2 siRNA.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay, siRNA knockdown, cell proliferation assay, genetic epistasis (siRNA rescue)\",\n      \"journal\": \"Neoplasia\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — direct interaction, in-cell ubiquitination, genetic epistasis rescue, multiple breast cancer cell lines\",\n      \"pmids\": [\"24027428\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"BCA2 (RNF115) acts as an E3 SUMO ligase for IκBα, enhancing IκBα SUMOylation, which sequesters NF-κB in the cytoplasm and reduces NF-κB-dependent HIV proviral transcription by up to 4.4-fold in CD4+ T cells. BCA2 is induced by NF-κB-activating cytokines and provides negative feedback on this pathway.\",\n      \"method\": \"In-cell SUMOylation assay, NF-κB reporter assay, siRNA knockdown, HIV transcription measurement in CD4+ T cells\",\n      \"journal\": \"Journal of virology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — functional SUMO ligase activity demonstrated in cells with mechanistic pathway placement, single lab\",\n      \"pmids\": [\"28122985\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF115 constitutively interacts with MAVS and catalyzes K48-linked ubiquitination and proteasomal degradation of MAVS in uninfected cells, maintaining basal MAVS levels. After HSV-1 infection, RNF115 associates with MITA/STING and catalyzes K63-linked ubiquitination to promote MITA aggregation and antiviral signaling. Rnf115-/- mice show elevated MAVS protein basally and impaired HSV-1-induced MITA aggregation.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay (K48 and K63 linkage-specific), Rnf115 knockout mice, MITA aggregation assay, viral infection models\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — dual substrate characterization with linkage-specific ubiquitination, in vivo knockout model with multiple phenotypic readouts, replicated across conditions\",\n      \"pmids\": [\"33139700\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF115 interacts with STX17 (syntaxin-17) and enhances its stability, which is required for autophagosome-lysosome fusion. RNF115 deletion causes accumulation of autophagosomes by impairing their fusion with lysosomes.\",\n      \"method\": \"Co-immunoprecipitation, autophagosome/lysosome fusion assay, RNF115 knockout cells, LC3/STX17 protein stability assay\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — co-IP with functional consequence in KO cells, single lab\",\n      \"pmids\": [\"32980859\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF115 mediates ubiquitination of p53, promoting its degradation in lung adenocarcinoma cells. RNF115 disruption induces G1 phase arrest and inhibits tumor growth in xenograft models.\",\n      \"method\": \"In vivo ubiquitination assay, siRNA/shRNA knockdown, cell cycle analysis, xenograft mouse model\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — ubiquitination assay plus in vivo functional data, single lab\",\n      \"pmids\": [\"32553631\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF115 promotes Wnt/β-catenin pathway activation by mediating ubiquitination and proteasomal degradation of APC (adenomatous polyposis coli) in lung adenocarcinoma cells.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, RNF115 knockdown/overexpression, β-catenin pathway reporter\",\n      \"journal\": \"Cancer & metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — ubiquitination of APC shown with functional Wnt pathway readout, single lab\",\n      \"pmids\": [\"33509267\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF115 is phosphorylated by AKT1, enabling 14-3-3 chaperones to bind phosphorylated RNF115 and facilitate its localization to the ER and Golgi apparatus. RNF115 then interacts with RAB1A and RAB13 and catalyzes K11-linked ubiquitination on specific lysine residues (Lys49/Lys61 of RAB1A; Lys46/Lys58 of RAB13), impairing GDI1 recruitment to RAB proteins and thereby blocking post-ER trafficking of Toll-like receptors.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay (K11-specific), site-directed mutagenesis, Rnf115 KO cells, subcellular fractionation/localization, TLR trafficking assay, AKT1 kinase assay\",\n      \"journal\": \"Advanced science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — linkage-specific ubiquitination with defined sites via mutagenesis, upstream kinase identified, downstream GDI1 mechanism, KO rescue experiments\",\n      \"pmids\": [\"35343654\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF115 is enriched on phagosomes of IFN-γ-activated macrophages and acts as a regulator of phagosomal maturation. Loss of RNF115 protein or its ligase activity enhances phagosomal maturation and increases cytokine responses to bacterial infection. RNF115 KO mice show less tissue damage in response to S. aureus infection.\",\n      \"method\": \"Quantitative proteomics of phagosomes, RNF115 KO (cells and mice), phagosomal maturation assay, bacterial infection assay, cytokine measurement\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — phagosomal proteomics + genetic KO with in vivo validation, multiple orthogonal methods\",\n      \"pmids\": [\"36281581\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF115 interacts with LC3B and downregulates LC3B protein levels, suppressing autophagy. RNF115 deletion promotes autophagy, reduces damaged mitochondria, and protects against LPS/D-GalN-induced acute liver injury in mice via NF-κB and JNK pathway inhibition.\",\n      \"method\": \"Co-immunoprecipitation (RNF115-LC3B), RNF115 KO mice, autophagy flux assay, adeno-associated virus rescue, pharmacological autophagy inhibition (3-MA), cytokine/macrophage analysis\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — co-IP with LC3B plus in vivo KO, but mechanistic detail of LC3B regulation is limited\",\n      \"pmids\": [\"38129372\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF115 ubiquitinates and promotes proteasomal degradation of CDK10, activating the Raf-1 pathway and promoting cell cycle progression in thyroid carcinoma cells. CDK10 overexpression rescues RNF115-induced malignant phenotypes.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, RNF115 KO/overexpression, CDK10 overexpression rescue, Raf-1 pathway analysis\",\n      \"journal\": \"Cell biology and toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — ubiquitination of CDK10 with pathway placement by rescue experiment, single lab\",\n      \"pmids\": [\"38376606\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Knockout or pharmacological inhibition of RNF115 (with disulfiram) impairs STING oligomerization and Golgi localization, thereby inhibiting STING-mediated inflammatory signaling. This effect is cell-type specific: RNF115 knockout inhibits STING activation in myeloid cells but not in endothelial cells or fibroblasts.\",\n      \"method\": \"RNF115 KO mice, bone marrow chimeric mice, disulfiram pharmacological inhibition, STING oligomerization assay, Golgi localization assay, PBMC cytokine measurement\",\n      \"journal\": \"Cellular & molecular immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo KO and pharmacological inhibition with mechanistic readout across multiple disease models\",\n      \"pmids\": [\"38267694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF115 acts as an E3 ubiquitin ligase for DHODH, promoting K27-linked ubiquitination of DHODH and inhibiting its autophagic degradation, thereby suppressing ferroptosis in hepatocellular carcinoma. YBX1 promotes RNF115 mRNA translation through m5C modification of the 3'-UTR.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay (K27-linkage), autophagic degradation assay, RNF115 KO/overexpression, RIP-seq, MeRIP-qPCR, dual-luciferase reporter\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — K27-ubiquitination of DHODH with functional ferroptosis readout; translational regulation by m5C is well-supported but single lab\",\n      \"pmids\": [\"40088428\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"RNF115 coordinates with the adaptor Tollip to mediate selective autophagic degradation of PEDV nucleocapsid (N) protein via K63-linked ubiquitination at residues K271, K276, K302, K359, and K375. The C25 and C38 residues of RNF115 are critical for N protein ubiquitination. Tollip forms a stable complex with RNF115 and N protein to facilitate substrate recognition.\",\n      \"method\": \"Co-immunoprecipitation, in vivo ubiquitination assay (K63-specific), site-directed mutagenesis, siRNA knockdown of Tollip, autophagic flux assay, viral replication assay\",\n      \"journal\": \"International journal of biological macromolecules\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — site-specific ubiquitination with mutagenesis and adaptor identification, single lab, recent paper\",\n      \"pmids\": [\"41587704\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF115 (BCA2/Rabring7) is a RING-H2-type E3 ubiquitin ligase (and E3 SUMO ligase) that uses its N-terminal BZF domain to bind ubiquitin/ubiquitinated proteins and its RING domain to catalyze autoubiquitination as well as K11-, K27-, K48-, and K63-linked ubiquitination of diverse substrates—including MAVS, MITA/STING, RAB1A, RAB13, p21, p53, APC, CDK10, EGFR, c-Myc, LC3B, and DHODH—thereby regulating innate antiviral and antibacterial immune responses, endolysosomal trafficking (via Rab7 interaction and tetherin cofactor activity), phagosomal maturation, autophagy, and cancer cell proliferation; its activity and localization are modulated by AKT1 phosphorylation and 14-3-3 binding, and by partner proteins such as hHR23a.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"RNF115 (BCA2/Rabring7) is a RING-H2-type E3 ubiquitin ligase that controls endolysosomal trafficking, innate immune signaling, autophagy, and cell proliferation by catalyzing K11-, K27-, K48-, and K63-linked ubiquitination of diverse substrates. Its N-terminal BCA2 zinc finger (BZF) domain binds ubiquitin and ubiquitinated proteins to recruit substrates, while the RING-H2 domain mediates catalytic activity with Ubc4/Ubc5 family E2 enzymes; AKT1 phosphorylation and 14-3-3 binding regulate its ER/Golgi localization and activity [PMID:18819927, PMID:17462600, PMID:35343654]. In innate immunity, RNF115 constitutively targets MAVS for K48-linked degradation to set basal levels, and upon viral infection switches to K63-linked ubiquitination of STING/MITA to promote its oligomerization and antiviral signaling, while on phagosomes it restrains phagosomal maturation during bacterial infection [PMID:33139700, PMID:38267694, PMID:36281581]. RNF115 also promotes cancer cell proliferation by ubiquitinating and degrading tumor suppressors p21 and p53, the Wnt pathway component APC, and CDK10, and regulates autophagy through interactions with STX17 and LC3B [PMID:24027428, PMID:32553631, PMID:33509267, PMID:38376606, PMID:32980859, PMID:38129372].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"The discovery that RNF115 binds GTP-Rab7 and is recruited to late endosomes/lysosomes established it as a regulator of endolysosomal trafficking, explaining why its overexpression causes perinuclear lysosome aggregation and impairs EGF degradation.\",\n      \"evidence\": \"GST pull-down, co-immunoprecipitation, LysoTracker imaging, and EGF degradation assay in mammalian cells\",\n      \"pmids\": [\"12972561\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Rab7 was not shown to be a direct ubiquitination substrate\", \"mechanism of lysosome aggregation was unclear\", \"physiological stoichiometry of the interaction was not determined\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Reconstitution of E3 ubiquitin ligase activity with Ubc4/Ubc5 E2 enzymes and identification of the catalytic Cys229 residue in the RING domain established RNF115 as a bona fide ubiquitin ligase that accelerates EGFR degradation.\",\n      \"evidence\": \"In vitro ubiquitination assay with recombinant proteins, C229S mutagenesis, EGFR degradation assay\",\n      \"pmids\": [\"17462600\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct EGFR ubiquitination by RNF115 was not shown\", \"ubiquitin chain linkage type was not determined\", \"relationship to c-Cbl was not resolved\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Mapping of the BZF domain as a ubiquitin-binding module and identification of autoubiquitination-regulatory lysines revealed a bipartite architecture in which substrate recognition (BZF) and catalysis (RING-H2) are separable functions.\",\n      \"evidence\": \"Yeast/bacterial two-hybrid, mutagenesis of K26/K32 (BZF) and K232/K260 (RING-proximal), in vivo ubiquitination assay\",\n      \"pmids\": [\"18819927\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Crystal structure of the BZF–ubiquitin complex was not determined\", \"how BZF-mediated ubiquitin binding directs substrate selectivity was unknown\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Demonstration that RNF115 cooperates with tetherin/BST-2 to internalize HIV-1 virions into CD63+ endosomes for lysosomal degradation revealed a direct antiviral effector function beyond general endosomal trafficking.\",\n      \"evidence\": \"Co-immunoprecipitation with tetherin, RNAi knockdown, viral particle release assay, confocal microscopy in HeLa cells\",\n      \"pmids\": [\"20019814\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether RNF115 ubiquitinates tetherin or viral proteins was not determined\", \"mechanism of virion internalization was unclear\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identification of hHR23a and 14-3-3σ as binding partners that stabilize RNF115 by suppressing autoubiquitination and by phosphorylation-dependent sequestration, respectively, revealed that RNF115 turnover is tightly regulated by opposing stabilization mechanisms.\",\n      \"evidence\": \"Yeast/bacterial two-hybrid, in vitro binding, in vivo ubiquitination assay with proteasome inhibition\",\n      \"pmids\": [\"22315970\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"The kinase responsible for RNF115 phosphorylation was not identified in this study\", \"in vivo relevance was not tested\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Establishing that RNF115 functions downstream of c-Cbl to ubiquitinate EGFR and promote multivesicular body sorting—and extends to MET and CXCR4—positioned it as a general regulator of receptor tyrosine kinase and GPCR endosomal sorting via ESCRT-II stabilization.\",\n      \"evidence\": \"Co-immunoprecipitation, siRNA knockdown, in vitro ubiquitination, electron microscopy of MVBs in HeLa cells\",\n      \"pmids\": [\"23418353\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct ubiquitination of ESCRT-II components was not tested\", \"structural basis for broad receptor substrate recognition was unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Discovery that RNF115 ubiquitinates p21Waf1/Cip1 via K48 linkage for proteasomal degradation, with genetic epistasis rescue, provided the first mechanistic link between RNF115 E3 activity and cell cycle control in breast cancer.\",\n      \"evidence\": \"Co-immunoprecipitation, K48-linkage-specific ubiquitination, p21 siRNA rescue of BCA2 knockdown growth arrest in breast cancer cell lines\",\n      \"pmids\": [\"24027428\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether RNF115-mediated p21 degradation occurs in non-transformed cells was unknown\", \"contribution relative to other p21 E3 ligases was not assessed\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Demonstration that RNF115 SUMOylates IκBα to sequester NF-κB in the cytoplasm established a dual E3 ligase function (ubiquitin and SUMO) and identified a negative feedback loop restraining NF-κB-driven HIV transcription in CD4+ T cells.\",\n      \"evidence\": \"In-cell SUMOylation assay, NF-κB reporter, siRNA knockdown, HIV transcription measurement in CD4+ T cells\",\n      \"pmids\": [\"28122985\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"SUMO ligase activity was not reconstituted in vitro with purified components\", \"whether this mechanism operates beyond HIV infection was untested\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Using Rnf115 knockout mice, the dual and context-dependent role of RNF115 in innate immunity was established: constitutive K48-linked ubiquitination of MAVS maintains basal signaling tone, while infection-induced K63-linked ubiquitination of STING/MITA drives antiviral defense.\",\n      \"evidence\": \"Co-immunoprecipitation, linkage-specific ubiquitination assays, Rnf115−/− mice, MITA aggregation assay, viral infection models\",\n      \"pmids\": [\"33139700\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Signal that triggers the MAVS-to-STING substrate switch was not identified\", \"whether other E3 ligases compensate in knockout animals was not addressed\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"The finding that RNF115 stabilizes STX17 to promote autophagosome–lysosome fusion, and that its loss causes autophagosome accumulation, extended RNF115's role from endosomal trafficking to autophagy regulation.\",\n      \"evidence\": \"Co-immunoprecipitation with STX17, RNF115 KO cells, autophagosome-lysosome fusion assay\",\n      \"pmids\": [\"32980859\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether STX17 is a direct ubiquitination substrate was not shown\", \"ubiquitin linkage type on STX17 was not determined\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identification of AKT1 as the upstream kinase that phosphorylates RNF115 to enable 14-3-3 binding and ER/Golgi relocalization, where RNF115 catalyzes K11-linked ubiquitination of RAB1A/RAB13 to block TLR post-ER trafficking, provided a complete signaling-to-substrate mechanism for immune regulation.\",\n      \"evidence\": \"AKT1 kinase assay, K11-linkage-specific ubiquitination, site-directed mutagenesis of RAB lysines, Rnf115 KO cells, subcellular fractionation, TLR trafficking assay\",\n      \"pmids\": [\"35343654\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether AKT1-mediated regulation applies to all RNF115 substrates was unknown\", \"structural basis for K11-linkage selectivity was not determined\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Phagosomal proteomics in IFN-γ-activated macrophages revealed RNF115 enrichment on phagosomes, with knockout showing enhanced phagosomal maturation and reduced tissue damage in S. aureus infection, establishing RNF115 as a brake on antimicrobial phagosome function.\",\n      \"evidence\": \"Quantitative phagosomal proteomics, RNF115 KO cells and mice, phagosomal maturation assay, S. aureus infection\",\n      \"pmids\": [\"36281581\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The phagosomal substrate(s) of RNF115 ligase activity were not identified\", \"mechanism distinguishing phagosomal from endosomal function was unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"RNF115 was shown to downregulate LC3B protein levels to suppress autophagy, and its deletion protected against LPS/D-GalN-induced acute liver injury in mice through reduced NF-κB and JNK activation, linking its autophagy-regulatory role to organ-level inflammatory damage.\",\n      \"evidence\": \"Co-immunoprecipitation with LC3B, RNF115 KO mice, autophagy flux assay, AAV rescue, 3-MA inhibition\",\n      \"pmids\": [\"38129372\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether LC3B is directly ubiquitinated by RNF115 and via which linkage was not defined\", \"relative contribution of autophagy versus NF-κB suppression to the protective phenotype was unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Demonstration that RNF115 knockout or pharmacological inhibition with disulfiram impairs STING oligomerization and Golgi localization specifically in myeloid cells refined the cell-type-specific model of RNF115-dependent innate immune control and suggested therapeutic tractability.\",\n      \"evidence\": \"RNF115 KO mice, bone marrow chimeras, disulfiram treatment, STING oligomerization and Golgi localization assays, PBMC cytokine measurements\",\n      \"pmids\": [\"38267694\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct target of disulfiram inhibition on RNF115 catalytic mechanism was not resolved\", \"basis for myeloid-cell-type specificity was unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identification of DHODH as a K27-linked ubiquitination substrate whose modification by RNF115 inhibits autophagic degradation and suppresses ferroptosis revealed a new linkage type and an unexpected role in metabolic cell death regulation in hepatocellular carcinoma.\",\n      \"evidence\": \"K27-linkage-specific ubiquitination assay, autophagic degradation assay, RNF115 KO/overexpression, RIP-seq and MeRIP-qPCR for m5C-dependent translational regulation\",\n      \"pmids\": [\"40088428\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"K27-ubiquitination mechanism was not reconstituted in vitro\", \"structural basis for linkage selectivity among K11, K27, K48, K63 across substrates remains unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RNF115 selects among K11, K27, K48, and K63 ubiquitin linkage types for different substrates, and what determines the context-dependent switch between constitutive degradative versus signal-amplifying ubiquitination programs, remain central unresolved questions.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No structural model of full-length RNF115 or its substrate complexes exists\", \"E2 enzyme pairing rules that dictate linkage specificity are unknown\", \"how AKT1 phosphorylation and 14-3-3 binding integrate with substrate-specific switches has not been resolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [1, 2, 6, 8, 10, 12, 13, 14, 17, 19, 20]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [1, 8, 10, 14, 19]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005764\", \"supporting_discovery_ids\": [0, 3, 6]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 4]},\n      {\"term_id\": \"GO:0005783\", \"supporting_discovery_ids\": [14]},\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [14]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005768\", \"supporting_discovery_ids\": [0, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [10, 14, 15, 18]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [1, 6, 8, 10, 14]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [0, 3, 6, 14]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [11, 16, 20]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [8, 12, 17]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [9, 13, 14]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [19]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"RAB7A\", \"MAVS\", \"TMEM173\", \"BST2\", \"RAB1A\", \"RAB13\", \"STX17\", \"TOLLIP\"],\n    \"other_free_text\": []\n  }\n}\n```"}