{"gene":"RNF114","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2008,"finding":"RNF114 (ZNF313) binds ubiquitin via an ubiquitin-interaction motif (UIM), demonstrated by cell-free ubiquitin-binding assays, identifying it as a ubiquitin-binding E3 ligase paralogue of TRAC-1.","method":"Cell-free ubiquitin-binding assay (UIM binding)","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — in vitro binding assay in a single study with no mutagenesis or structural validation reported in abstract","pmids":["18364390"],"is_preprint":false},{"year":2011,"finding":"RNF114 is a soluble cytosolic protein that associates with ubiquitinated proteins, is induced by interferons and synthetic dsRNA, and its over-expression enhances NF-κB and IRF3 reporter activity and increases type I and type III IFN mRNA levels, indicating it positively regulates a dsRNA-induced innate immune feedback loop.","method":"Subcellular fractionation, reporter assays (NF-κB/IRF3), mRNA quantification, siRNA knockdown/overexpression","journal":"Human molecular genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (fractionation, reporter, mRNA levels) in a single lab","pmids":["21571784"],"is_preprint":false},{"year":2013,"finding":"ZNF313/RNF114 is an E3 ubiquitin ligase that ubiquitinates and destabilizes p21(WAF1), p27(KIP1), and p57(KIP2) via its RING domain, thereby promoting G1-to-S cell cycle progression and suppressing cellular senescence; RING domain mutations abolish this activity.","method":"In vitro ubiquitination assay, RING domain mutagenesis, co-immunoprecipitation, protein stability assays, cell cycle analysis","journal":"Cell death and differentiation","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus active-site mutagenesis and multiple cellular readouts, single lab","pmids":["23645206"],"is_preprint":false},{"year":2013,"finding":"ZNF313/RNF114 shows both nuclear and cytoplasmic localization in normal epithelial cells but predominantly cytoplasmic distribution in carcinoma cells, as determined by immunohistochemistry and subcellular localization experiments.","method":"Immunohistochemistry, subcellular localization","journal":"Cell death and differentiation","confidence":"Low","confidence_rationale":"Tier 3 / Weak — descriptive localization data without direct functional consequence established in the same experiment","pmids":["23645206"],"is_preprint":false},{"year":2014,"finding":"RNF114 interacts with A20 in T cells (identified by two-hybrid screening and confirmed by co-immunoprecipitation), modulates A20 ubiquitylation, stabilizes A20 and IκBα proteins, and thereby acts as a negative regulator of NF-κB-dependent transcription, modulating T-cell activation and apoptosis.","method":"Yeast two-hybrid screening, co-immunoprecipitation, ubiquitylation assay, NF-κB reporter assay, T-cell activation assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, reporter assay, and functional T-cell readouts in a single lab","pmids":["25165885"],"is_preprint":false},{"year":2014,"finding":"XAF1 activates ZNF313/RNF114-mediated ubiquitination of p21(WAF1) to terminate p53-dependent cell-cycle arrest, and XAF1 interacts with ZNF313 through its zinc finger domain 7 (ZF7); truncated XAF1 isoforms lacking ZF7 cannot activate ZNF313.","method":"Co-immunoprecipitation, ubiquitination assays, domain deletion mapping, reporter assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP domain mapping and functional ubiquitination assays, single lab","pmids":["25313037"],"is_preprint":false},{"year":2014,"finding":"RNF114 promotes T cell activation in a dose-dependent manner; the two C2H2 domains play opposing roles (upstream C2H2 suppresses, downstream C2H2 enhances T cell activation); 23 RNF114-interacting proteins were identified by tandem affinity purification/mass spectrometry.","method":"Flow cytometry (FACS), tandem affinity purification (TAP), mass spectrometry, domain deletion constructs","journal":"Immunobiology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — domain deletion with phenotypic readout but limited mechanistic follow-up; interactome from single pull-down experiment","pmids":["24631332"],"is_preprint":false},{"year":2017,"finding":"RNF114 is an E3 ubiquitin ligase that negatively regulates RLH/MAVS-mediated type I IFN production; MAVS is a substrate for RNF114-mediated polyubiquitination and degradation; RNF114 knockout mice exhibit increased basal IFN levels and sensitized responses to dsRNA.","method":"In vitro ubiquitination assay, co-immunoprecipitation, RNF114 knockout mouse model, IFN measurement","journal":"Cytokine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vitro ubiquitination plus KO mouse with IFN phenotype, single lab","pmids":["28625874"],"is_preprint":false},{"year":2017,"finding":"RNF114 ubiquitinates and degrades TAB1 via the ubiquitin-proteasome pathway in mouse oocytes; RNF114-mediated TAB1 degradation activates the NF-κB pathway and is required for maternal-to-zygotic transition (MZT); knockdown of Rnf114 arrests embryo development at the two-cell stage.","method":"Protein microarray substrate screening (9,000 proteins), in vitro ubiquitination, mouse oocyte knockdown, embryo development assay","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — substrate identified by unbiased protein microarray, validated by in vitro ubiquitination, and confirmed by in vivo knockdown with developmental phenotype","pmids":["28073917"],"is_preprint":false},{"year":2017,"finding":"RNF114 suppresses RANKL-induced osteoclastogenesis by promoting K48-linked proteasomal degradation of TRAF6 and suppressing RANKL-activated NFATc1 and NFAT-regulated promoter activity, acting upstream of TAK1/TAB2 in the RANK/TRAF6/NF-κB pathway.","method":"RNF114 knockout mouse osteoclast differentiation, ectopic expression, TRAP activity assay, bone resorption assay, NF-κB/NFAT reporter assay, protein stability assay","journal":"Journal of orthopaedic research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO and overexpression with multiple cellular readouts and epistasis placement, single lab","pmids":["28708287"],"is_preprint":false},{"year":2020,"finding":"Sea perch LjRNF114 targets MAVS and TRAF3 for K27- and K48-linked ubiquitination and proteasomal degradation, suppressing IFN production; the RING and UIM domains of RNF114 are required for interaction with TRAF3 and MAVS.","method":"Co-immunoprecipitation, domain mapping, ubiquitination assay, IFN reporter assay, overexpression/knockdown","journal":"Journal of immunology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mapping with functional ubiquitination assays; non-mammalian ortholog study with mechanistic detail","pmids":["33268485"],"is_preprint":false},{"year":2020,"finding":"Porcine RNF114 interacts with PRRSV nonstructural protein Nsp12 and promotes K27-linked polyubiquitination and proteasome-dependent degradation of Nsp12, suppressing viral replication; this activity requires RNF114 ubiquitin ligase activity.","method":"Co-immunoprecipitation, ubiquitination assay, viral titer assay, knockdown/overexpression, proteasome inhibitor","journal":"Veterinary microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, in vitro ubiquitination, and viral replication readout; single lab","pmids":["32605740"],"is_preprint":false},{"year":2021,"finding":"Covalent small molecules that react with RNF114 at Cys8 can be used as PROTAC E3 ligase recruiters to degrade therapeutic targets (BRD4, BCR-ABL) in cells, demonstrating that RNF114 can be engaged as an E3 ligase in targeted protein degradation.","method":"Activity-based protein profiling, covalent ligand screening, PROTAC synthesis and cellular degradation assays","journal":"Cell chemical biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — chemoproteomic identification plus cell-based PROTAC degradation with multiple targets, single lab","pmids":["33513350"],"is_preprint":false},{"year":2021,"finding":"Maternal RNF114 ubiquitinates and degrades CBX5 (HP1α) in mouse oocytes; accumulation of CBX5 or TAB1 in RNF114-depleted two-cell embryos impedes major zygotic genome activation; knockdown of either CBX5 or TAB1 partially rescues the developmental arrest caused by RNF114 depletion.","method":"Maternal protein depletion, immunofluorescence, transcriptome analysis, in vitro ubiquitination, rescue experiments","journal":"Development","confidence":"High","confidence_rationale":"Tier 1 / Moderate — substrate identification, in vitro ubiquitination, and genetic rescue with transcriptome confirmation, single lab","pmids":["34104941"],"is_preprint":false},{"year":2022,"finding":"Human RNF114 interacts with MAVS and inhibits type I interferon production, thereby promoting VSV replication; overexpression promotes and depletion reduces viral replication both in vitro and in vivo.","method":"Co-immunoprecipitation, overexpression/knockdown, viral replication assays, in vivo mouse experiment","journal":"Microbial pathogenesis","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — co-IP and functional viral replication assays, single lab, in vivo confirmation","pmids":["35545202"],"is_preprint":false},{"year":2022,"finding":"RNF114 mediates ubiquitylation and degradation of EGR1, as demonstrated by ubiquitylation experiments in gastric cancer cells.","method":"Ubiquitylation assay, knockdown, western blot","journal":"Journal of Cancer","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, limited mechanistic follow-up described in abstract","pmids":["35069903"],"is_preprint":false},{"year":2023,"finding":"RNF114 is a PARylation-dependent E3 ubiquitin ligase recruited to DNA lesions in a PAR-dependent manner, where it targets PARP1 for K48-linked ubiquitination and degradation; blockade of RNF114 (by nimbolide, which inhibits its E3 activity) causes PARP1 trapping at DNA lesions and synthetic lethality with BRCA mutations.","method":"Co-immunoprecipitation, in vitro ubiquitination, laser-induced DNA damage recruitment imaging, nimbolide-RNF114 inhibition, cellular viability assays, in vivo xenograft","journal":"Science advances","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple orthogonal methods (co-IP, in vitro ubiquitination, live imaging, small-molecule inhibition, in vivo), single lab but comprehensive","pmids":["37878693"],"is_preprint":false},{"year":2023,"finding":"XAF1 acts as an adaptor for VCP-mediated deubiquitination of RNF114, which activates RNF114 to promote K48-linked ubiquitination and degradation of junction plakoglobin (JUP), facilitating colorectal cancer cell migration and metastasis.","method":"Co-immunoprecipitation, ubiquitination assay, VCP inhibition, migration/invasion assay, in vivo metastasis model","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP, ubiquitination assay, and in vivo metastasis with mechanistic cascade established, single lab","pmids":["38095639"],"is_preprint":false},{"year":2024,"finding":"XAF1 facilitates ZNF313/RNF114-mediated K48-linked polyubiquitination and proteasomal degradation of TRIM28 by interacting with the RING domain of ZNF313 through XAF1's ZF7 domain, thereby suppressing TRIM28 oncogenic activity; XAF1 mutants lacking ZF7 cannot promote TRIM28 ubiquitination.","method":"Co-immunoprecipitation, ubiquitination assay, domain deletion mapping, tumor growth assays","journal":"Molecular biomedicine","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — co-IP with domain mapping and functional ubiquitination assays, single lab","pmids":["39532800"],"is_preprint":false},{"year":2025,"finding":"RNF114 preferentially recognizes ubiquitinated ADP-ribose (ADPr-Ub / MARUbe) over non-modified ubiquitin, and extends K11-linked polyubiquitin chains on MARUbylated substrates; the tandem ZnF2+ZnF3+UIM (Di19-UIM) domain module is required for recognizing MARUbylated species and for RNF114 recruitment to sites of laser-induced DNA damage.","method":"Non-hydrolysable ADPr-Ub probe, proteomics pull-down, biophysical binding assays (biochemical), domain deletion analysis, laser-induced DNA damage recruitment imaging, in vitro K11 ubiquitin chain assembly","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1 / Strong — novel chemical probe, proteomics identification, biophysical validation, domain mutagenesis, and cell-based localization; independently corroborated by parallel EMBO Journal paper","pmids":["40634336"],"is_preprint":false},{"year":2025,"finding":"RNF114 (and RNF166) act as reader-writer E3 ligases that recognize the MARUbe hybrid mark on PARP7 (generated by DTX2) through a tandem Di19-UIM module (M-UBD) and extend K11-linked polyubiquitin chains at those sites; this places RNF114 in a PARP7 MARUbylation-K11 polyUb pathway.","method":"Fluorescent Ub-ADPr chemoenzymatic probe, in vitro ubiquitination, AlphaFold3 structural modeling, cellular PARP7 MARUbylation assays","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 / Strong — chemical probe, in vitro reconstitution, structural modeling, and cellular assays; replicates and extends findings of PMID 40634336","pmids":["41039157"],"is_preprint":false},{"year":2025,"finding":"RNF114 and RNF166 recognize a monoubiquitin-MAR hybrid mark on tankyrase (generated by Deltex ligases DTX2/DTX3) via a hybrid reader domain containing two binding sites (one for MAR, one for ubiquitin), and further diubiquitylate this mark; this blocks PAR formation and antagonizes RNF146-mediated tankyrase degradation, stabilizing tankyrase.","method":"Cellular ubiquitylation assays, immunoprecipitation, domain mapping","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — cellular and biochemical assays in preprint, single lab, not yet peer-reviewed","pmids":[],"is_preprint":true},{"year":2025,"finding":"RNF114 promotes K48-linked polyubiquitination and proteasomal degradation of PRRSV nsp12 at lysine residues 89, 91, 127, and 130; mutations at these lysines enable PRRSV to evade RNF114-mediated degradation and enhance viral replication.","method":"Co-immunoprecipitation, ubiquitination assay, site-directed mutagenesis of nsp12 lysines, reverse genetics viral replication assay, mass spectrometry","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — lysine mutagenesis and reverse genetics with viral replication phenotype; preprint, not yet peer-reviewed","pmids":[],"is_preprint":true},{"year":2025,"finding":"SPP1 interacts with RNF114 and facilitates RNF114-mediated ubiquitination of P85α, promoting PI3K/AKT/mTOR signaling and NSCLC brain metastasis.","method":"Co-immunoprecipitation, LC-MS, shRNA knockdown, in vitro/in vivo metastasis assays","journal":"Molecular carcinogenesis","confidence":"Low","confidence_rationale":"Tier 3 / Weak — co-IP and knockdown phenotype, limited ubiquitination mechanistic detail in abstract, single lab","pmids":["39918025"],"is_preprint":false},{"year":2010,"finding":"The ZNF313 promoter has a functional Sp1 binding site within the fragment from nt -157 to +8, confirmed by site-directed mutation and mithramycin A treatment, identifying Sp1 as a transcriptional regulator of RNF114.","method":"Promoter deletion constructs, luciferase reporter assay, site-directed mutagenesis, mithramycin A treatment","journal":"Cell biology international","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — promoter mutagenesis and chemical inhibition confirm Sp1 site; single lab","pmids":["20515446"],"is_preprint":false}],"current_model":"RNF114 is a RING-domain E3 ubiquitin ligase and ubiquitin-binding protein that uses a tandem zinc finger–UIM (Di19-UIM / M-UBD) module to read the hybrid MARUbe (mono-ADP-ribose–ubiquitin) mark on substrates and extend K11-linked polyubiquitin chains; it ubiquitinates and degrades multiple substrates—including PARP1 (PAR-dependent, at DNA damage sites), p21/p27/p57 CIP/KIP CDK inhibitors, TAB1, TRAF6, MAVS, CBX5, TRIM28, and viral proteins (PRRSV Nsp12)—to regulate cell cycle progression, maternal-to-zygotic transition, innate immune/IFN signaling, NF-κB activity, osteoclastogenesis, and DNA damage response, while its activity is itself modulated by interactors such as XAF1, A20, and VCP."},"narrative":{"mechanistic_narrative":"RNF114 (ZNF313) is a RING-domain E3 ubiquitin ligase and ubiquitin-binding protein that couples recognition of ubiquitin and ADP-ribose marks to targeted substrate ubiquitination, regulating cell-cycle progression, the maternal-to-zygotic transition, innate immune signaling, and the DNA damage response [PMID:23645206, PMID:28073917, PMID:37878693]. Through its RING domain it ubiquitinates and destabilizes the CIP/KIP CDK inhibitors p21, p27, and p57 to drive G1-to-S progression and suppress senescence [PMID:23645206], and this activity toward p21 is licensed by XAF1, which engages RNF114 through its ZF7 zinc finger [PMID:25313037]. In early development, maternal RNF114 degrades TAB1 and the heterochromatin factor CBX5/HP1α, a step required for major zygotic genome activation and progression past the two-cell stage [PMID:28073917, PMID:34104941]. In the DNA damage response, RNF114 is recruited to lesions in a PARylation-dependent manner and ubiquitinates PARP1 for K48-linked degradation, such that inhibiting its ligase activity with nimbolide traps PARP1 on chromatin and produces synthetic lethality in BRCA-mutant cells [PMID:37878693]. The structural basis of this recruitment is a tandem ZnF2–ZnF3–UIM (Di19-UIM) module that preferentially reads the hybrid mono-ADP-ribose–ubiquitin (MARUbe) mark and extends K11-linked polyubiquitin chains on MARUbylated substrates such as PARP7 [PMID:40634336, PMID:41039157]. RNF114 also acts on innate immune signaling, both positively—as an interferon- and dsRNA-inducible enhancer of NF-κB/IRF3-driven IFN responses [PMID:21571784]—and negatively, by targeting MAVS for polyubiquitination and degradation, with RNF114-knockout mice showing elevated basal IFN [PMID:28625874]. Its NF-κB output is further shaped by stabilization of A20 [PMID:25165885] and degradation of TRAF6 during osteoclastogenesis [PMID:28708287]. RNF114 can be pharmacologically engaged as a PROTAC-recruiting E3 via a reactive Cys8 [PMID:33513350].","teleology":[{"year":2008,"claim":"Established that RNF114 is not merely a RING ligase but also a ubiquitin reader, raising the question of how ubiquitin recognition feeds into its catalytic output.","evidence":"Cell-free ubiquitin-binding assay of the UIM","pmids":["18364390"],"confidence":"Medium","gaps":["No mutagenesis or structural validation of the UIM-ubiquitin interface","Functional consequence of ubiquitin binding not yet linked to substrate selection"]},{"year":2010,"claim":"Defined a transcriptional input to RNF114 expression by mapping a functional Sp1 site in its promoter.","evidence":"Promoter deletion, luciferase reporter, site-directed mutagenesis and mithramycin A in cells","pmids":["20515446"],"confidence":"Medium","gaps":["Does not address signal-dependent regulation of the promoter","No link between Sp1 control and RNF114 protein activity"]},{"year":2011,"claim":"Positioned RNF114 within antiviral innate immunity as an interferon/dsRNA-inducible positive feedback factor for IFN production.","evidence":"Subcellular fractionation, NF-κB/IRF3 reporters, IFN mRNA quantification, siRNA/overexpression","pmids":["21571784"],"confidence":"Medium","gaps":["No substrate identified to explain the positive effect","Reconciliation with later negative regulation of MAVS not addressed"]},{"year":2013,"claim":"Provided the first reconstituted enzymatic role by showing RING-dependent ubiquitination and degradation of CIP/KIP inhibitors p21/p27/p57, linking RNF114 to cell-cycle progression.","evidence":"In vitro ubiquitination, RING mutagenesis, co-IP, stability and cell-cycle assays","pmids":["23645206"],"confidence":"High","gaps":["Localization shifts in carcinoma described only descriptively (#3)","Chain linkage type on CIP/KIP substrates not defined"]},{"year":2014,"claim":"Identified upstream regulators and refined the NF-κB role, showing XAF1 (via ZF7) activates p21 ubiquitination while A20 stabilization makes RNF114 a negative NF-κB regulator in T cells.","evidence":"Yeast two-hybrid, reciprocal co-IP, domain mapping, ubiquitination and reporter/T-cell assays","pmids":["25313037","25165885","24631332"],"confidence":"Medium","gaps":["Opposing pro- and anti-inflammatory effects context-dependence unresolved","Functional role of the 23 TAP/MS interactors largely uncharacterized"]},{"year":2017,"claim":"Defined RNF114 as essential for the maternal-to-zygotic transition by degrading TAB1, and as a negative regulator of MAVS-driven IFN and of TRAF6 in osteoclastogenesis.","evidence":"Protein microarray substrate screen, in vitro ubiquitination, oocyte knockdown and embryo assays; KO mouse IFN phenotype; KO osteoclast differentiation","pmids":["28073917","28625874","28708287"],"confidence":"High","gaps":["How a single ligase balances positive (#1) and negative (#7) IFN effects unresolved","Chain linkages differ across substrates without unifying recognition logic at this stage"]},{"year":2021,"claim":"Showed that maternal RNF114 also degrades CBX5/HP1α with TAB1 to permit zygotic genome activation, and that RNF114 is chemically tractable as a PROTAC E3 via Cys8.","evidence":"Maternal depletion with rescue and transcriptomics; chemoproteomics and cellular PROTAC degradation","pmids":["34104941","33513350"],"confidence":"High","gaps":["Endogenous ligand/regulation of Cys8 reactivity unknown","Structural basis of substrate selection still undefined at this point"]},{"year":2023,"claim":"Connected RNF114 to the DNA damage response, showing PAR-dependent recruitment and K48 ubiquitination of PARP1, with therapeutic implications for BRCA-mutant cancers.","evidence":"Co-IP, in vitro ubiquitination, laser-damage recruitment imaging, nimbolide inhibition, viability and xenograft assays","pmids":["37878693"],"confidence":"High","gaps":["Did not resolve the molecular reader for the PAR signal","Selectivity of PARP1 over other PARylated targets undefined"]},{"year":2025,"claim":"Resolved the mechanistic reader, establishing that a tandem Di19-UIM (M-UBD) module recognizes the hybrid MARUbe mark and that RNF114 extends K11-linked chains on MARUbylated substrates such as PARP7, defining it as a reader-writer ligase.","evidence":"Non-hydrolysable ADPr-Ub probes, proteomics, biophysical binding, domain mutagenesis, laser-damage imaging, in vitro K11 chain assembly, AlphaFold3 modeling","pmids":["40634336","41039157"],"confidence":"High","gaps":["Full structural model of substrate engagement not experimentally solved","How K11 extension dictates substrate fate beyond degradation unclear"]},{"year":null,"claim":"It remains unresolved how RNF114's single reader-writer module is partitioned across its many substrates and opposing pathway outcomes (IFN, cell cycle, development, metastasis) within a given cell.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model linking MARUbe recognition to non-PAR substrates (p21, TAB1, CBX5, TRAF6)","Tissue- and context-specific regulation of substrate choice uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[2,8,16,19]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[2,8,16]},{"term_id":"GO:0140313","term_label":"molecular sequestering activity","supporting_discovery_ids":[0,19]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[3,16]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[2]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[16,19]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[1,7]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[2,8,13]},{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[8,13]}],"complexes":[],"partners":["XAF1","A20","MAVS","TAB1","TRAF6","VCP","TRIM28","PARP1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9Y508","full_name":"E3 ubiquitin-protein ligase RNF114","aliases":["RING finger protein 114","RING-type E3 ubiquitin transferase RNF114","Zinc finger protein 228","Zinc finger protein 313"],"length_aa":228,"mass_kda":25.7,"function":"E3 ubiquitin-protein ligase that promotes the ubiquitination of various substrates (PubMed:23645206, PubMed:25165885). In turn, participates in the regulation of many biological processes including cell cycle, apoptosis, osteoclastogenesis as well as innate or adaptive immunity (PubMed:25165885, PubMed:28708287). Acts as a negative regulator of NF-kappa-B-dependent transcription by promoting the ubiquitination and stabilization of the NF-kappa-B inhibitor TNFAIP3 (PubMed:25165885). May promote the ubiquitination of TRAF6 as well (PubMed:28708287). Also acts as a negative regulator of T-cell activation (PubMed:25165885). Inhibits cellular dsRNA responses and interferon production by targeting MAVS component for proteasomal degradation (PubMed:25165885). Ubiquitinates the CDK inhibitor CDKN1A leading to its degradationand probably also CDKN1B and CDKN1C (PubMed:23645206). This activity stimulates cell cycle G1-to-S phase transition and suppresses cellular senescence. May play a role in spermatogenesis","subcellular_location":"Cytoplasm; Nucleus","url":"https://www.uniprot.org/uniprotkb/Q9Y508/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF114","classification":"Not Classified","n_dependent_lines":3,"n_total_lines":1208,"dependency_fraction":0.0024834437086092716},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF114","total_profiled":1310},"omim":[{"mim_id":"612950","title":"PSORIASIS 12, SUSCEPTIBILITY TO; PSORS12","url":"https://www.omim.org/entry/612950"},{"mim_id":"612451","title":"RING FINGER PROTEIN 114; RNF114","url":"https://www.omim.org/entry/612451"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Plasma membrane","reliability":"Supported"},{"location":"Cytosol","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"testis","ntpm":153.8}],"url":"https://www.proteinatlas.org/search/RNF114"},"hgnc":{"alias_symbol":["PSORS12"],"prev_symbol":["ZNF313"]},"alphafold":{"accession":"Q9Y508","domains":[{"cath_id":"3.30.40.10","chopping":"24-109","consensus_level":"high","plddt":94.4203,"start":24,"end":109},{"cath_id":"-","chopping":"137-208","consensus_level":"medium","plddt":86.9607,"start":137,"end":208}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y508","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y508-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9Y508-F1-predicted_aligned_error_v6.png","plddt_mean":83.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF114","jax_strain_url":"https://www.jax.org/strain/search?query=RNF114"},"sequence":{"accession":"Q9Y508","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9Y508.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9Y508/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9Y508"}},"corpus_meta":[{"pmid":"18364390","id":"PMC_18364390","title":"Identification of ZNF313/RNF114 as a novel psoriasis susceptibility gene.","date":"2008","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/18364390","citation_count":159,"is_preprint":false},{"pmid":"33513350","id":"PMC_33513350","title":"Chemoproteomics-enabled discovery of covalent RNF114-based degraders that mimic natural product function.","date":"2021","source":"Cell chemical biology","url":"https://pubmed.ncbi.nlm.nih.gov/33513350","citation_count":136,"is_preprint":false},{"pmid":"21571784","id":"PMC_21571784","title":"Functional analysis of the RNF114 psoriasis susceptibility gene implicates innate immune responses to double-stranded RNA in disease pathogenesis.","date":"2011","source":"Human molecular genetics","url":"https://pubmed.ncbi.nlm.nih.gov/21571784","citation_count":70,"is_preprint":false},{"pmid":"23645206","id":"PMC_23645206","title":"ZNF313 is a novel cell cycle activator with an E3 ligase activity inhibiting cellular senescence by destabilizing p21(WAF1.).","date":"2013","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/23645206","citation_count":62,"is_preprint":false},{"pmid":"25313037","id":"PMC_25313037","title":"XAF1 directs apoptotic switch of p53 signaling through activation of HIPK2 and ZNF313.","date":"2014","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/25313037","citation_count":61,"is_preprint":false},{"pmid":"28073917","id":"PMC_28073917","title":"The E3 ubiquitin ligase RNF114 and TAB1 degradation are required for maternal-to-zygotic transition.","date":"2017","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/28073917","citation_count":55,"is_preprint":false},{"pmid":"25165885","id":"PMC_25165885","title":"The RING ubiquitin E3 RNF114 interacts with A20 and modulates NF-κB activity and T-cell activation.","date":"2014","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/25165885","citation_count":50,"is_preprint":false},{"pmid":"37878693","id":"PMC_37878693","title":"Nimbolide targets RNF114 to induce the trapping of PARP1 and synthetic lethality in BRCA-mutated cancer.","date":"2023","source":"Science advances","url":"https://pubmed.ncbi.nlm.nih.gov/37878693","citation_count":47,"is_preprint":false},{"pmid":"33268485","id":"PMC_33268485","title":"E3 Ubiquitin Ligase RNF114 Inhibits Innate Immune Response to Red-Spotted Grouper Nervous Necrosis Virus Infection in Sea Perch by Targeting MAVS and TRAF3 to Mediate Their Degradation.","date":"2020","source":"Journal of immunology (Baltimore, Md. : 1950)","url":"https://pubmed.ncbi.nlm.nih.gov/33268485","citation_count":36,"is_preprint":false},{"pmid":"32605740","id":"PMC_32605740","title":"Proteasomal degradation of nonstructural protein 12 by RNF114 suppresses porcine reproductive and respiratory syndrome virus 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England)","url":"https://pubmed.ncbi.nlm.nih.gov/34104941","citation_count":17,"is_preprint":false},{"pmid":"38095639","id":"PMC_38095639","title":"XAF1 promotes colorectal cancer metastasis via VCP-RNF114-JUP axis.","date":"2023","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/38095639","citation_count":16,"is_preprint":false},{"pmid":"24631332","id":"PMC_24631332","title":"Identification of RNF114 as a novel positive regulatory protein for T cell activation.","date":"2014","source":"Immunobiology","url":"https://pubmed.ncbi.nlm.nih.gov/24631332","citation_count":16,"is_preprint":false},{"pmid":"35069903","id":"PMC_35069903","title":"RNF114 Silencing Inhibits the Proliferation and Metastasis of Gastric Cancer.","date":"2022","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/35069903","citation_count":14,"is_preprint":false},{"pmid":"12621547","id":"PMC_12621547","title":"Identification of a novel human zinc finger protein gene ZNF313.","date":"2003","source":"Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica","url":"https://pubmed.ncbi.nlm.nih.gov/12621547","citation_count":14,"is_preprint":false},{"pmid":"28708287","id":"PMC_28708287","title":"Regulation of RANKL-induced osteoclastogenesis by RING finger protein RNF114.","date":"2017","source":"Journal of orthopaedic research : official publication of the Orthopaedic Research Society","url":"https://pubmed.ncbi.nlm.nih.gov/28708287","citation_count":13,"is_preprint":false},{"pmid":"17394778","id":"PMC_17394778","title":"Cloning and expression analysis of a novel mouse zinc finger protein gene Znf313 abundantly expressed in testis.","date":"2007","source":"Journal of biochemistry and molecular biology","url":"https://pubmed.ncbi.nlm.nih.gov/17394778","citation_count":11,"is_preprint":false},{"pmid":"25290666","id":"PMC_25290666","title":"A RING finger protein 114 (RNF114) homolog from Chinese sturgeon (Acipenser sinensis) possesses immune-regulation properties via modulating RIG-I signaling pathway-mediated interferon expression.","date":"2014","source":"Fish & shellfish immunology","url":"https://pubmed.ncbi.nlm.nih.gov/25290666","citation_count":11,"is_preprint":false},{"pmid":"35545202","id":"PMC_35545202","title":"E3 ubiquitin ligase RNF114 promotes vesicular stomatitis virus replication via inhibiting type I interferon production.","date":"2022","source":"Microbial pathogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/35545202","citation_count":8,"is_preprint":false},{"pmid":"41039157","id":"PMC_41039157","title":"RNF114 and RNF166 exemplify reader-writer E3 ligases that extend K11 polyubiquitin onto sites of MARUbylation.","date":"2025","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/41039157","citation_count":7,"is_preprint":false},{"pmid":"37523804","id":"PMC_37523804","title":"RNF114 facilitates the proliferation, stemness, and metastasis of colorectal cancer.","date":"2023","source":"Pathology, research and practice","url":"https://pubmed.ncbi.nlm.nih.gov/37523804","citation_count":7,"is_preprint":false},{"pmid":"37855431","id":"PMC_37855431","title":"DCAF13 and RNF114 participate in the regulation of early porcine embryo development.","date":"2023","source":"Reproduction (Cambridge, England)","url":"https://pubmed.ncbi.nlm.nih.gov/37855431","citation_count":5,"is_preprint":false},{"pmid":"39532800","id":"PMC_39532800","title":"XAF1 antagonizes TRIM28 activity through the assembly of a ZNF313-mediated destruction complex to suppress tumor malignancy.","date":"2024","source":"Molecular biomedicine","url":"https://pubmed.ncbi.nlm.nih.gov/39532800","citation_count":4,"is_preprint":false},{"pmid":"39918025","id":"PMC_39918025","title":"SPP1 Promotes NSCLC Brain Metastasis Via Sequestration of Ubiquitin Ligase RNF114 to Facilitate P85α Ubiquitination.","date":"2025","source":"Molecular carcinogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/39918025","citation_count":2,"is_preprint":false},{"pmid":"40092691","id":"PMC_40092691","title":"RNF114 Interacts with EWSR1 to Regulate VEGFR2 in HER2-positive Breast Cancer.","date":"2025","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/40092691","citation_count":1,"is_preprint":false},{"pmid":"20515446","id":"PMC_20515446","title":"Sp1 plays an important role in regulating the transcription of ZNF313.","date":"2010","source":"Cell biology international","url":"https://pubmed.ncbi.nlm.nih.gov/20515446","citation_count":1,"is_preprint":false},{"pmid":"41046487","id":"PMC_41046487","title":"Emerging role of the E3 ubiquitin ligase RNF114 in health and disease.","date":"2025","source":"Human cell","url":"https://pubmed.ncbi.nlm.nih.gov/41046487","citation_count":0,"is_preprint":false},{"pmid":"40981192","id":"PMC_40981192","title":"Computational Identification of RNF114 nsSNPs with Potential Roles in Psoriasis and Immune Dysregulation.","date":"2025","source":"Medical sciences (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/40981192","citation_count":0,"is_preprint":false},{"pmid":"41039156","id":"PMC_41039156","title":"RNF114, a RING E3 ligase that reads and extends the hybrid ADP-ribose-ubiquitin signal.","date":"2025","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/41039156","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.05.11.653360","title":"A family of E3 ligases extend K11 polyubiquitin on sites of MARUbylation","date":"2025-05-15","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.11.653360","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.09.02.673672","title":"Adaptive mutations at lysine residues of PRRSV nsp12 enable evasion of host proteasomal and selective autophagic 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Vivo","date":"2025-04-08","source":"bioRxiv","url":"https://doi.org/10.1101/2025.04.07.647692","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":19209,"output_tokens":5780,"usd":0.072164,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":14285,"output_tokens":3599,"usd":0.0807,"stage2_stop_reason":"end_turn"},"total_usd":0.152864,"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\": \"RNF114 (ZNF313) binds ubiquitin via an ubiquitin-interaction motif (UIM), demonstrated by cell-free ubiquitin-binding assays, identifying it as a ubiquitin-binding E3 ligase paralogue of TRAC-1.\",\n      \"method\": \"Cell-free ubiquitin-binding assay (UIM binding)\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — in vitro binding assay in a single study with no mutagenesis or structural validation reported in abstract\",\n      \"pmids\": [\"18364390\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"RNF114 is a soluble cytosolic protein that associates with ubiquitinated proteins, is induced by interferons and synthetic dsRNA, and its over-expression enhances NF-κB and IRF3 reporter activity and increases type I and type III IFN mRNA levels, indicating it positively regulates a dsRNA-induced innate immune feedback loop.\",\n      \"method\": \"Subcellular fractionation, reporter assays (NF-κB/IRF3), mRNA quantification, siRNA knockdown/overexpression\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (fractionation, reporter, mRNA levels) in a single lab\",\n      \"pmids\": [\"21571784\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ZNF313/RNF114 is an E3 ubiquitin ligase that ubiquitinates and destabilizes p21(WAF1), p27(KIP1), and p57(KIP2) via its RING domain, thereby promoting G1-to-S cell cycle progression and suppressing cellular senescence; RING domain mutations abolish this activity.\",\n      \"method\": \"In vitro ubiquitination assay, RING domain mutagenesis, co-immunoprecipitation, protein stability assays, cell cycle analysis\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro ubiquitination reconstitution plus active-site mutagenesis and multiple cellular readouts, single lab\",\n      \"pmids\": [\"23645206\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"ZNF313/RNF114 shows both nuclear and cytoplasmic localization in normal epithelial cells but predominantly cytoplasmic distribution in carcinoma cells, as determined by immunohistochemistry and subcellular localization experiments.\",\n      \"method\": \"Immunohistochemistry, subcellular localization\",\n      \"journal\": \"Cell death and differentiation\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — descriptive localization data without direct functional consequence established in the same experiment\",\n      \"pmids\": [\"23645206\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RNF114 interacts with A20 in T cells (identified by two-hybrid screening and confirmed by co-immunoprecipitation), modulates A20 ubiquitylation, stabilizes A20 and IκBα proteins, and thereby acts as a negative regulator of NF-κB-dependent transcription, modulating T-cell activation and apoptosis.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation, ubiquitylation assay, NF-κB reporter assay, T-cell activation assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, reporter assay, and functional T-cell readouts in a single lab\",\n      \"pmids\": [\"25165885\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"XAF1 activates ZNF313/RNF114-mediated ubiquitination of p21(WAF1) to terminate p53-dependent cell-cycle arrest, and XAF1 interacts with ZNF313 through its zinc finger domain 7 (ZF7); truncated XAF1 isoforms lacking ZF7 cannot activate ZNF313.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, domain deletion mapping, reporter assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP domain mapping and functional ubiquitination assays, single lab\",\n      \"pmids\": [\"25313037\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RNF114 promotes T cell activation in a dose-dependent manner; the two C2H2 domains play opposing roles (upstream C2H2 suppresses, downstream C2H2 enhances T cell activation); 23 RNF114-interacting proteins were identified by tandem affinity purification/mass spectrometry.\",\n      \"method\": \"Flow cytometry (FACS), tandem affinity purification (TAP), mass spectrometry, domain deletion constructs\",\n      \"journal\": \"Immunobiology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — domain deletion with phenotypic readout but limited mechanistic follow-up; interactome from single pull-down experiment\",\n      \"pmids\": [\"24631332\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RNF114 is an E3 ubiquitin ligase that negatively regulates RLH/MAVS-mediated type I IFN production; MAVS is a substrate for RNF114-mediated polyubiquitination and degradation; RNF114 knockout mice exhibit increased basal IFN levels and sensitized responses to dsRNA.\",\n      \"method\": \"In vitro ubiquitination assay, co-immunoprecipitation, RNF114 knockout mouse model, IFN measurement\",\n      \"journal\": \"Cytokine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vitro ubiquitination plus KO mouse with IFN phenotype, single lab\",\n      \"pmids\": [\"28625874\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RNF114 ubiquitinates and degrades TAB1 via the ubiquitin-proteasome pathway in mouse oocytes; RNF114-mediated TAB1 degradation activates the NF-κB pathway and is required for maternal-to-zygotic transition (MZT); knockdown of Rnf114 arrests embryo development at the two-cell stage.\",\n      \"method\": \"Protein microarray substrate screening (9,000 proteins), in vitro ubiquitination, mouse oocyte knockdown, embryo development assay\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — substrate identified by unbiased protein microarray, validated by in vitro ubiquitination, and confirmed by in vivo knockdown with developmental phenotype\",\n      \"pmids\": [\"28073917\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"RNF114 suppresses RANKL-induced osteoclastogenesis by promoting K48-linked proteasomal degradation of TRAF6 and suppressing RANKL-activated NFATc1 and NFAT-regulated promoter activity, acting upstream of TAK1/TAB2 in the RANK/TRAF6/NF-κB pathway.\",\n      \"method\": \"RNF114 knockout mouse osteoclast differentiation, ectopic expression, TRAP activity assay, bone resorption assay, NF-κB/NFAT reporter assay, protein stability assay\",\n      \"journal\": \"Journal of orthopaedic research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO and overexpression with multiple cellular readouts and epistasis placement, single lab\",\n      \"pmids\": [\"28708287\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Sea perch LjRNF114 targets MAVS and TRAF3 for K27- and K48-linked ubiquitination and proteasomal degradation, suppressing IFN production; the RING and UIM domains of RNF114 are required for interaction with TRAF3 and MAVS.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping, ubiquitination assay, IFN reporter assay, overexpression/knockdown\",\n      \"journal\": \"Journal of immunology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mapping with functional ubiquitination assays; non-mammalian ortholog study with mechanistic detail\",\n      \"pmids\": [\"33268485\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Porcine RNF114 interacts with PRRSV nonstructural protein Nsp12 and promotes K27-linked polyubiquitination and proteasome-dependent degradation of Nsp12, suppressing viral replication; this activity requires RNF114 ubiquitin ligase activity.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, viral titer assay, knockdown/overexpression, proteasome inhibitor\",\n      \"journal\": \"Veterinary microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, in vitro ubiquitination, and viral replication readout; single lab\",\n      \"pmids\": [\"32605740\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Covalent small molecules that react with RNF114 at Cys8 can be used as PROTAC E3 ligase recruiters to degrade therapeutic targets (BRD4, BCR-ABL) in cells, demonstrating that RNF114 can be engaged as an E3 ligase in targeted protein degradation.\",\n      \"method\": \"Activity-based protein profiling, covalent ligand screening, PROTAC synthesis and cellular degradation assays\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — chemoproteomic identification plus cell-based PROTAC degradation with multiple targets, single lab\",\n      \"pmids\": [\"33513350\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Maternal RNF114 ubiquitinates and degrades CBX5 (HP1α) in mouse oocytes; accumulation of CBX5 or TAB1 in RNF114-depleted two-cell embryos impedes major zygotic genome activation; knockdown of either CBX5 or TAB1 partially rescues the developmental arrest caused by RNF114 depletion.\",\n      \"method\": \"Maternal protein depletion, immunofluorescence, transcriptome analysis, in vitro ubiquitination, rescue experiments\",\n      \"journal\": \"Development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — substrate identification, in vitro ubiquitination, and genetic rescue with transcriptome confirmation, single lab\",\n      \"pmids\": [\"34104941\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Human RNF114 interacts with MAVS and inhibits type I interferon production, thereby promoting VSV replication; overexpression promotes and depletion reduces viral replication both in vitro and in vivo.\",\n      \"method\": \"Co-immunoprecipitation, overexpression/knockdown, viral replication assays, in vivo mouse experiment\",\n      \"journal\": \"Microbial pathogenesis\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — co-IP and functional viral replication assays, single lab, in vivo confirmation\",\n      \"pmids\": [\"35545202\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF114 mediates ubiquitylation and degradation of EGR1, as demonstrated by ubiquitylation experiments in gastric cancer cells.\",\n      \"method\": \"Ubiquitylation assay, knockdown, western blot\",\n      \"journal\": \"Journal of Cancer\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, limited mechanistic follow-up described in abstract\",\n      \"pmids\": [\"35069903\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF114 is a PARylation-dependent E3 ubiquitin ligase recruited to DNA lesions in a PAR-dependent manner, where it targets PARP1 for K48-linked ubiquitination and degradation; blockade of RNF114 (by nimbolide, which inhibits its E3 activity) causes PARP1 trapping at DNA lesions and synthetic lethality with BRCA mutations.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination, laser-induced DNA damage recruitment imaging, nimbolide-RNF114 inhibition, cellular viability assays, in vivo xenograft\",\n      \"journal\": \"Science advances\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple orthogonal methods (co-IP, in vitro ubiquitination, live imaging, small-molecule inhibition, in vivo), single lab but comprehensive\",\n      \"pmids\": [\"37878693\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"XAF1 acts as an adaptor for VCP-mediated deubiquitination of RNF114, which activates RNF114 to promote K48-linked ubiquitination and degradation of junction plakoglobin (JUP), facilitating colorectal cancer cell migration and metastasis.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, VCP inhibition, migration/invasion assay, in vivo metastasis model\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP, ubiquitination assay, and in vivo metastasis with mechanistic cascade established, single lab\",\n      \"pmids\": [\"38095639\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"XAF1 facilitates ZNF313/RNF114-mediated K48-linked polyubiquitination and proteasomal degradation of TRIM28 by interacting with the RING domain of ZNF313 through XAF1's ZF7 domain, thereby suppressing TRIM28 oncogenic activity; XAF1 mutants lacking ZF7 cannot promote TRIM28 ubiquitination.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, domain deletion mapping, tumor growth assays\",\n      \"journal\": \"Molecular biomedicine\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — co-IP with domain mapping and functional ubiquitination assays, single lab\",\n      \"pmids\": [\"39532800\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF114 preferentially recognizes ubiquitinated ADP-ribose (ADPr-Ub / MARUbe) over non-modified ubiquitin, and extends K11-linked polyubiquitin chains on MARUbylated substrates; the tandem ZnF2+ZnF3+UIM (Di19-UIM) domain module is required for recognizing MARUbylated species and for RNF114 recruitment to sites of laser-induced DNA damage.\",\n      \"method\": \"Non-hydrolysable ADPr-Ub probe, proteomics pull-down, biophysical binding assays (biochemical), domain deletion analysis, laser-induced DNA damage recruitment imaging, in vitro K11 ubiquitin chain assembly\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — novel chemical probe, proteomics identification, biophysical validation, domain mutagenesis, and cell-based localization; independently corroborated by parallel EMBO Journal paper\",\n      \"pmids\": [\"40634336\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF114 (and RNF166) act as reader-writer E3 ligases that recognize the MARUbe hybrid mark on PARP7 (generated by DTX2) through a tandem Di19-UIM module (M-UBD) and extend K11-linked polyubiquitin chains at those sites; this places RNF114 in a PARP7 MARUbylation-K11 polyUb pathway.\",\n      \"method\": \"Fluorescent Ub-ADPr chemoenzymatic probe, in vitro ubiquitination, AlphaFold3 structural modeling, cellular PARP7 MARUbylation assays\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — chemical probe, in vitro reconstitution, structural modeling, and cellular assays; replicates and extends findings of PMID 40634336\",\n      \"pmids\": [\"41039157\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF114 and RNF166 recognize a monoubiquitin-MAR hybrid mark on tankyrase (generated by Deltex ligases DTX2/DTX3) via a hybrid reader domain containing two binding sites (one for MAR, one for ubiquitin), and further diubiquitylate this mark; this blocks PAR formation and antagonizes RNF146-mediated tankyrase degradation, stabilizing tankyrase.\",\n      \"method\": \"Cellular ubiquitylation assays, immunoprecipitation, domain mapping\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — cellular and biochemical assays in preprint, single lab, not yet peer-reviewed\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"RNF114 promotes K48-linked polyubiquitination and proteasomal degradation of PRRSV nsp12 at lysine residues 89, 91, 127, and 130; mutations at these lysines enable PRRSV to evade RNF114-mediated degradation and enhance viral replication.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, site-directed mutagenesis of nsp12 lysines, reverse genetics viral replication assay, mass spectrometry\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — lysine mutagenesis and reverse genetics with viral replication phenotype; preprint, not yet peer-reviewed\",\n      \"pmids\": [],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"SPP1 interacts with RNF114 and facilitates RNF114-mediated ubiquitination of P85α, promoting PI3K/AKT/mTOR signaling and NSCLC brain metastasis.\",\n      \"method\": \"Co-immunoprecipitation, LC-MS, shRNA knockdown, in vitro/in vivo metastasis assays\",\n      \"journal\": \"Molecular carcinogenesis\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — co-IP and knockdown phenotype, limited ubiquitination mechanistic detail in abstract, single lab\",\n      \"pmids\": [\"39918025\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"The ZNF313 promoter has a functional Sp1 binding site within the fragment from nt -157 to +8, confirmed by site-directed mutation and mithramycin A treatment, identifying Sp1 as a transcriptional regulator of RNF114.\",\n      \"method\": \"Promoter deletion constructs, luciferase reporter assay, site-directed mutagenesis, mithramycin A treatment\",\n      \"journal\": \"Cell biology international\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — promoter mutagenesis and chemical inhibition confirm Sp1 site; single lab\",\n      \"pmids\": [\"20515446\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF114 is a RING-domain E3 ubiquitin ligase and ubiquitin-binding protein that uses a tandem zinc finger–UIM (Di19-UIM / M-UBD) module to read the hybrid MARUbe (mono-ADP-ribose–ubiquitin) mark on substrates and extend K11-linked polyubiquitin chains; it ubiquitinates and degrades multiple substrates—including PARP1 (PAR-dependent, at DNA damage sites), p21/p27/p57 CIP/KIP CDK inhibitors, TAB1, TRAF6, MAVS, CBX5, TRIM28, and viral proteins (PRRSV Nsp12)—to regulate cell cycle progression, maternal-to-zygotic transition, innate immune/IFN signaling, NF-κB activity, osteoclastogenesis, and DNA damage response, while its activity is itself modulated by interactors such as XAF1, A20, and VCP.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RNF114 (ZNF313) is a RING-domain E3 ubiquitin ligase and ubiquitin-binding protein that couples recognition of ubiquitin and ADP-ribose marks to targeted substrate ubiquitination, regulating cell-cycle progression, the maternal-to-zygotic transition, innate immune signaling, and the DNA damage response [#2, #8, #16]. Through its RING domain it ubiquitinates and destabilizes the CIP/KIP CDK inhibitors p21, p27, and p57 to drive G1-to-S progression and suppress senescence [#2], and this activity toward p21 is licensed by XAF1, which engages RNF114 through its ZF7 zinc finger [#5]. In early development, maternal RNF114 degrades TAB1 and the heterochromatin factor CBX5/HP1\\u03b1, a step required for major zygotic genome activation and progression past the two-cell stage [#8, #13]. In the DNA damage response, RNF114 is recruited to lesions in a PARylation-dependent manner and ubiquitinates PARP1 for K48-linked degradation, such that inhibiting its ligase activity with nimbolide traps PARP1 on chromatin and produces synthetic lethality in BRCA-mutant cells [#16]. The structural basis of this recruitment is a tandem ZnF2\\u2013ZnF3\\u2013UIM (Di19-UIM) module that preferentially reads the hybrid mono-ADP-ribose\\u2013ubiquitin (MARUbe) mark and extends K11-linked polyubiquitin chains on MARUbylated substrates such as PARP7 [#19, #20]. RNF114 also acts on innate immune signaling, both positively\\u2014as an interferon- and dsRNA-inducible enhancer of NF-\\u03baB/IRF3-driven IFN responses [#1]\\u2014and negatively, by targeting MAVS for polyubiquitination and degradation, with RNF114-knockout mice showing elevated basal IFN [#7]. Its NF-\\u03baB output is further shaped by stabilization of A20 [#4] and degradation of TRAF6 during osteoclastogenesis [#9]. RNF114 can be pharmacologically engaged as a PROTAC-recruiting E3 via a reactive Cys8 [#12].\",\n  \"teleology\": [\n    {\n      \"year\": 2008,\n      \"claim\": \"Established that RNF114 is not merely a RING ligase but also a ubiquitin reader, raising the question of how ubiquitin recognition feeds into its catalytic output.\",\n      \"evidence\": \"Cell-free ubiquitin-binding assay of the UIM\",\n      \"pmids\": [\"18364390\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mutagenesis or structural validation of the UIM-ubiquitin interface\", \"Functional consequence of ubiquitin binding not yet linked to substrate selection\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined a transcriptional input to RNF114 expression by mapping a functional Sp1 site in its promoter.\",\n      \"evidence\": \"Promoter deletion, luciferase reporter, site-directed mutagenesis and mithramycin A in cells\",\n      \"pmids\": [\"20515446\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not address signal-dependent regulation of the promoter\", \"No link between Sp1 control and RNF114 protein activity\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Positioned RNF114 within antiviral innate immunity as an interferon/dsRNA-inducible positive feedback factor for IFN production.\",\n      \"evidence\": \"Subcellular fractionation, NF-\\u03baB/IRF3 reporters, IFN mRNA quantification, siRNA/overexpression\",\n      \"pmids\": [\"21571784\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No substrate identified to explain the positive effect\", \"Reconciliation with later negative regulation of MAVS not addressed\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Provided the first reconstituted enzymatic role by showing RING-dependent ubiquitination and degradation of CIP/KIP inhibitors p21/p27/p57, linking RNF114 to cell-cycle progression.\",\n      \"evidence\": \"In vitro ubiquitination, RING mutagenesis, co-IP, stability and cell-cycle assays\",\n      \"pmids\": [\"23645206\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Localization shifts in carcinoma described only descriptively (#3)\", \"Chain linkage type on CIP/KIP substrates not defined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identified upstream regulators and refined the NF-\\u03baB role, showing XAF1 (via ZF7) activates p21 ubiquitination while A20 stabilization makes RNF114 a negative NF-\\u03baB regulator in T cells.\",\n      \"evidence\": \"Yeast two-hybrid, reciprocal co-IP, domain mapping, ubiquitination and reporter/T-cell assays\",\n      \"pmids\": [\"25313037\", \"25165885\", \"24631332\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Opposing pro- and anti-inflammatory effects context-dependence unresolved\", \"Functional role of the 23 TAP/MS interactors largely uncharacterized\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Defined RNF114 as essential for the maternal-to-zygotic transition by degrading TAB1, and as a negative regulator of MAVS-driven IFN and of TRAF6 in osteoclastogenesis.\",\n      \"evidence\": \"Protein microarray substrate screen, in vitro ubiquitination, oocyte knockdown and embryo assays; KO mouse IFN phenotype; KO osteoclast differentiation\",\n      \"pmids\": [\"28073917\", \"28625874\", \"28708287\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How a single ligase balances positive (#1) and negative (#7) IFN effects unresolved\", \"Chain linkages differ across substrates without unifying recognition logic at this stage\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Showed that maternal RNF114 also degrades CBX5/HP1\\u03b1 with TAB1 to permit zygotic genome activation, and that RNF114 is chemically tractable as a PROTAC E3 via Cys8.\",\n      \"evidence\": \"Maternal depletion with rescue and transcriptomics; chemoproteomics and cellular PROTAC degradation\",\n      \"pmids\": [\"34104941\", \"33513350\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Endogenous ligand/regulation of Cys8 reactivity unknown\", \"Structural basis of substrate selection still undefined at this point\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Connected RNF114 to the DNA damage response, showing PAR-dependent recruitment and K48 ubiquitination of PARP1, with therapeutic implications for BRCA-mutant cancers.\",\n      \"evidence\": \"Co-IP, in vitro ubiquitination, laser-damage recruitment imaging, nimbolide inhibition, viability and xenograft assays\",\n      \"pmids\": [\"37878693\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve the molecular reader for the PAR signal\", \"Selectivity of PARP1 over other PARylated targets undefined\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Resolved the mechanistic reader, establishing that a tandem Di19-UIM (M-UBD) module recognizes the hybrid MARUbe mark and that RNF114 extends K11-linked chains on MARUbylated substrates such as PARP7, defining it as a reader-writer ligase.\",\n      \"evidence\": \"Non-hydrolysable ADPr-Ub probes, proteomics, biophysical binding, domain mutagenesis, laser-damage imaging, in vitro K11 chain assembly, AlphaFold3 modeling\",\n      \"pmids\": [\"40634336\", \"41039157\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Full structural model of substrate engagement not experimentally solved\", \"How K11 extension dictates substrate fate beyond degradation unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how RNF114's single reader-writer module is partitioned across its many substrates and opposing pathway outcomes (IFN, cell cycle, development, metastasis) within a given cell.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model linking MARUbe recognition to non-PAR substrates (p21, TAB1, CBX5, TRAF6)\", \"Tissue- and context-specific regulation of substrate choice uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [2, 8, 16, 19]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [2, 8, 16]},\n      {\"term_id\": \"GO:0140313\", \"supporting_discovery_ids\": [0, 19]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [3, 16]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [16, 19]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [1, 7]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 8, 13]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [8, 13]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"XAF1\", \"A20\", \"MAVS\", \"TAB1\", \"TRAF6\", \"VCP\", \"TRIM28\", \"PARP1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":7,"faith_total":8,"faith_pct":87.5}}