{"gene":"RNF144B","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2010,"finding":"IBRDC2 (RNF144B) is an IBR-type RING-finger E3 ubiquitin ligase that regulates Bax stability through ubiquitination-dependent degradation, protecting cells from spontaneous apoptosis. Upon apoptosis induction, IBRDC2 translocates from cytosol to mitochondrial domains enriched with active Bax, physically interacts with activated Bax, and this translocation requires both mitochondrial localization and apoptotic activation of Bax, and depends on Bcl-xL expression levels.","method":"Co-immunoprecipitation (physical interaction with activated Bax), active Bax-specific antibody staining, Bax mutant analysis in HCT116 Bax(-/-) cells, ubiquitination assays with IBRDC2 overexpression/knockdown, subcellular fractionation/live imaging","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, Bax mutant epistasis in defined Bax-null cells, multiple orthogonal methods (ubiquitination assay, fractionation, active Bax antibody), single rigorous study with robust controls","pmids":["20300062"],"is_preprint":false},{"year":2003,"finding":"p53RFP (RNF144B) is a transcriptional target of p53 with E3 ubiquitin ligase activity that interacts with and ubiquitinates p21WAF1, promoting its degradation. Its expression is negatively correlated with p21WAF1 protein levels, suggesting it regulates p21WAF1 stability.","method":"E3 ubiquitin ligase activity assay, co-immunoprecipitation (p53RFP–p21WAF1 interaction), expression correlation analysis, transcriptional reporter assay for p53 induction","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — E3 ligase activity demonstrated in vitro, Co-IP interaction shown, single lab with two orthogonal methods","pmids":["12853982"],"is_preprint":false},{"year":2006,"finding":"p53RFP (RNF144B) induces p53-dependent but caspase-independent apoptosis. Its RING-IBR-RING domain mediates interaction with E2 ubiquitin-conjugating enzymes UbcH7 and UbcH8 but not UbcH5. The conserved C-terminal domain (not the E3 ligase domain) is required and sufficient for apoptosis induction, indicating apoptosis does not require E3 ubiquitin ligase activity.","method":"Co-immunoprecipitation (E2 enzyme binding), domain deletion mutant analysis, apoptosis assays (caspase-independent), overexpression in cell lines","journal":"FEBS letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain mutant analysis combined with Co-IP for E2 binding specificity, single lab with two orthogonal methods","pmids":["16427630"],"is_preprint":false},{"year":2012,"finding":"PIR2/RNF144B is a direct transcriptional target of ΔNp63α in keratinocytes. It binds and mediates proteasomal degradation of both p21WAF1/CIP1 and ΔNp63α itself, forming an auto-regulatory feedback loop. Depletion of PIR2/RNF144B impairs keratinocyte proliferation and differentiation with accumulation of p21WAF1/CIP1.","method":"Transcriptional reporter assay (ΔNp63α target), co-immunoprecipitation (PIR2–ΔNp63α binding), proteasome inhibitor rescue, siRNA knockdown with proliferation/differentiation phenotypic readout","journal":"Oncogene","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal binding shown by Co-IP, proteasomal degradation confirmed with inhibitor, loss-of-function phenotype with defined molecular readout, multiple orthogonal methods in one study","pmids":["23128396"],"is_preprint":false},{"year":2019,"finding":"RNF144B interacts with TBK1 through its IBR domain binding the scaffold/dimerization domain (SDD) of TBK1, inhibiting TBK1 phosphorylation and K63-linked polyubiquitination, which leads to TBK1 inactivation, IRF3 dephosphorylation, and reduced IFN-β production in response to LPS stimulation. RNF144B knockdown increases IRF3 activation and IFN-β production.","method":"Co-immunoprecipitation (RNF144B–TBK1 interaction), domain mapping (IBR domain–SDD interaction), siRNA knockdown, Western blot for phosphorylation and ubiquitination status, IFN-β ELISA","journal":"Journal of leukocyte biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP with domain mapping, siRNA knockdown with defined signaling readouts, single lab with multiple orthogonal methods","pmids":["31509299"],"is_preprint":false},{"year":2016,"finding":"RNF144B is necessary for priming of inflammasome responses in primary human macrophages; specifically, it promotes LPS-inducible IL-1β mRNA expression but does not regulate several other LPS-inducible cytokines (e.g., IL-10, IFN-γ) or inflammasome components (procaspase-1, pro-IL-18). RNF144B is LPS-inducible in human but not mouse macrophages due to differences in transcription factor binding sites in the promoter.","method":"Gene silencing (siRNA) in primary human macrophages, cytokine mRNA quantification (qPCR), cap analysis of gene expression (CAGE) for promoter activity, comparative analysis across human and mouse cell types","journal":"Journal of leukocyte biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — gene silencing with specific cytokine readouts, CAGE promoter analysis, multiple cell types compared; single lab","pmids":["26819317"],"is_preprint":false},{"year":2018,"finding":"PIR2/RNF144B protein is stabilized via phosphorylation downstream of GSK3β activity, and this stabilization is necessary for proliferation of endometrial cancer cells in the absence of oestrogenic growth stimuli. Inactivation of GSK3β leads to loss of PIR2/RNF144B protein and consequent inhibition of cell proliferation.","method":"GSK3β inhibitor treatment, phosphorylation analysis, siRNA knockdown, cell proliferation assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — pharmacological GSK3β inhibition linked to RNF144B stability loss and proliferation; single lab with two orthogonal methods but no direct demonstration of phosphorylation site","pmids":["29724995"],"is_preprint":false},{"year":2022,"finding":"RNF144B interacts with FCER2 in human spermatogonial stem cells (SSCs), and through FCER2 activates the NOTCH2/HES1 pathway (FCER2 pulls down NOTCH2 intracellular domain N2ICD). RNF144B knockdown reduces NOTCH2, FCER2, HES1, and HEY1 levels, inhibits SSC proliferation, and promotes apoptosis; these effects are reversed by FCER2 overexpression.","method":"RNA sequencing (target identification), co-immunoprecipitation (RNF144B–FCER2 and FCER2–N2ICD interactions), siRNA knockdown, FCER2 overexpression rescue, proliferation and apoptosis assays","journal":"Journal of cellular physiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP interactions, epistasis rescue experiment with FCER2 overexpression, RNA-seq target identification; single lab with multiple orthogonal methods","pmids":["35699595"],"is_preprint":false},{"year":2023,"finding":"RNF144B interacts with TBK1; in sepsis model, Rnf144b deficiency results in impaired TBK1 activation but enhanced NF-κB activation in macrophages. Conditional knockout of Rnf144b in myeloid cells leads to increased inflammatory cytokines after LPS/CpG stimulation, higher mortality, and exacerbated cardiac dysfunction in septic mice.","method":"Conditional knockout mice (myeloid-specific), co-immunoprecipitation (Rnf144b–TBK1), Western blot for TBK1 and NF-κB activation, cytokine measurement, in vivo sepsis model","journal":"ESC heart failure","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — conditional KO mice with defined in vivo phenotype, Co-IP, signaling readouts; single lab","pmids":["37088470"],"is_preprint":false},{"year":2023,"finding":"HDAC3 promotes RNF144B expression in ovarian cancer cells. RNF144B interacts with p21 and regulates degradation of the p21/p53 complex. RNF144B-mediated p21 degradation (assessed by cycloheximide chase) promotes ovarian cancer cell proliferation, migration, and invasion while inhibiting apoptosis.","method":"Co-immunoprecipitation (RNF144B/p21/p53 interaction), cycloheximide chase (p21 stability), transcriptome profiling, siRNA knockdown and overexpression, in vivo tumor model","journal":"Tissue & cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP interaction, CHX chase for substrate stability, in vivo validation; single lab","pmids":["37992458"],"is_preprint":false},{"year":2024,"finding":"RNF144B specifically interacts with MDA5 and promotes K27/K33-linked polyubiquitination of MDA5 at lysine residues K23 and K43 (within the CARDs domain), which promotes autophagic degradation of MDA5 by p62. Rnf144b knockout in mice greatly promotes IFN production, inhibits EMCV replication, and significantly increases overall survival upon EMCV infection.","method":"Co-immunoprecipitation (RNF144B–MDA5 interaction), ubiquitination assay with linkage type and site identification (K27/K33, K23/K43), Rnf144b knockout mice, viral challenge (EMCV), IFN production assay, autophagy pathway analysis","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — ubiquitination site mapping, linkage-type determination, KO mouse in vivo phenotype with viral infection, multiple orthogonal methods including site-specific mutagenesis implied by lysine identification","pmids":["39285245"],"is_preprint":false},{"year":2024,"finding":"RNF144B acts as a tumor suppressor downstream of TP53 in lung adenocarcinoma, mediating protein degradation associated with cell cycle progression, DNA damage response, and genomic stability. RNF144B deficiency induces chromosomal instability, mitotic defects, and resistance to cell cycle inhibitors that induce chromosomal instability.","method":"RNF144B knockout in human and mouse cells (non-transformed and cancerous), proteomics and transcriptomics analysis, proliferation/transformation assays, cell cycle analysis, DNA damage response assays, in vivo tumor models","journal":"Journal of experimental & clinical cancer research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO models in multiple cell types, proteomics and transcriptomics with functional validation; single lab","pmids":["38685100"],"is_preprint":false},{"year":2026,"finding":"RNF144B interacts with TRAF3 and promotes its K48-linked ubiquitination and proteasomal degradation. In the absence of RNF144B (KO mice), TRAF3 is stabilized, leading to enhanced NF-κB and MAPK signaling pathway activation and exacerbated neuroinflammation after ischemic stroke. TRAF3 knockdown in RNF144B-deficient mice partially reversed neurological dysfunction and neuroinflammation post-MCAO.","method":"Co-immunoprecipitation (RNF144B–TRAF3 interaction), ubiquitination assay (K48-linked), RNF144B knockout mice (MCAO model), TRAF3 knockdown epistasis rescue, Western blot for NF-κB and MAPK activation, immunofluorescence","journal":"Biochemical pharmacology","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — Co-IP, K48-linked ubiquitination demonstrated, KO mouse in vivo phenotype, epistasis rescue with TRAF3 knockdown, multiple orthogonal methods","pmids":["41903812"],"is_preprint":false}],"current_model":"RNF144B (also known as IBRDC2/p53RFP/PIR2) is a RING-IBR-RING domain E3 ubiquitin ligase that is transcriptionally induced by p53 and ΔNp63α, and acts as a multi-substrate ubiquitin ligase that targets p21WAF1, ΔNp63α, Bax, MDA5, and TRAF3 for ubiquitination-dependent degradation; it interacts with E2 enzymes UbcH7/UbcH8 via its RING-IBR-RING domain, is stabilized by GSK3β-dependent phosphorylation, translocates to mitochondria upon apoptosis to interact with activated Bax, suppresses innate immune signaling by binding TBK1 (via its IBR domain) to inhibit TBK1 phosphorylation/K63-ubiquitination and by promoting K27/K33-ubiquitination of MDA5 at K23/K43 to direct its autophagic degradation via p62, degrades TRAF3 via K48-linked ubiquitination to restrain NF-κB/MAPK signaling, and functions as a tumor suppressor maintaining genomic stability downstream of TP53."},"narrative":{"mechanistic_narrative":"RNF144B is a RING-IBR-RING (RBR) E3 ubiquitin ligase that operates downstream of the p53/p63 transcription factor network to control cell fate and innate immune signaling [PMID:12853982, PMID:38685100]. It is a direct transcriptional target of p53 and of ΔNp63α, and its RING-IBR-RING domain engages the E2 conjugating enzymes UbcH7 and UbcH8 to drive substrate ubiquitination [PMID:12853982, PMID:16427630, PMID:23128396]. As a multi-substrate ligase, RNF144B promotes degradation of the cyclin-dependent kinase inhibitor p21WAF1/CIP1 and of ΔNp63α itself, the latter forming an auto-regulatory feedback loop required for normal keratinocyte proliferation and differentiation [PMID:12853982, PMID:23128396]. In apoptosis, RNF144B ubiquitinates Bax to restrain spontaneous cell death, and upon apoptotic stimulation it translocates from the cytosol to mitochondrial domains enriched in activated Bax [PMID:20300062]. RNF144B is a recurrent negative regulator of innate immune signaling: it binds TBK1 through its IBR domain to suppress TBK1 phosphorylation, K63-ubiquitination, and downstream IRF3/IFN-β activation; it directs K27/K33-linked ubiquitination of MDA5 at K23/K43 within the CARDs to route MDA5 for p62-dependent autophagic degradation, limiting antiviral interferon responses; and it drives K48-linked ubiquitination and proteasomal degradation of TRAF3 to restrain NF-κB and MAPK signaling [PMID:31509299, PMID:39285245, PMID:41903812]. Acting downstream of TP53, RNF144B functions as a tumor suppressor that maintains genomic stability, with its loss producing chromosomal instability and mitotic defects [PMID:38685100].","teleology":[{"year":2003,"claim":"Established RNF144B as a p53-inducible E3 ligase, linking it to the tumor suppressor transcriptional program and identifying p21WAF1 as a degradation substrate.","evidence":"In vitro E3 ligase assay, Co-IP of p53RFP-p21WAF1, p53 transcriptional reporter","pmids":["12853982"],"confidence":"Medium","gaps":["E2 partner and ubiquitin linkage type not defined","in vivo relevance of p21 turnover untested"]},{"year":2006,"claim":"Mapped E2 enzyme specificity (UbcH7/UbcH8, not UbcH5) to the RING-IBR-RING domain and showed RNF144B-induced apoptosis is caspase-independent and separable from its ligase activity.","evidence":"Co-IP for E2 binding, domain deletion mutants, apoptosis assays in cell lines","pmids":["16427630"],"confidence":"Medium","gaps":["substrate ubiquitinated during apoptosis not identified","mechanism of caspase-independent death by the C-terminal domain unresolved"]},{"year":2010,"claim":"Defined RNF144B as a regulator of Bax stability that protects against spontaneous apoptosis and relocalizes to mitochondria to engage activated Bax during cell death.","evidence":"Reciprocal Co-IP with activated Bax, Bax mutant epistasis in HCT116 Bax-null cells, ubiquitination assays, subcellular fractionation/live imaging","pmids":["20300062"],"confidence":"High","gaps":["ubiquitin linkage type on Bax not defined","trigger for cytosol-to-mitochondria translocation not mechanistically resolved"]},{"year":2012,"claim":"Placed RNF144B in a ΔNp63α auto-regulatory feedback loop in keratinocytes, showing it degrades both p21WAF1 and ΔNp63α to enable proliferation and differentiation.","evidence":"ΔNp63α transcriptional reporter, reciprocal Co-IP, proteasome inhibitor rescue, siRNA knockdown with proliferation/differentiation readout","pmids":["23128396"],"confidence":"High","gaps":["ubiquitin linkage on ΔNp63α not characterized","in vivo epidermal phenotype untested"]},{"year":2016,"claim":"Implicated RNF144B in inflammasome priming, showing it is LPS-inducible in human (not mouse) macrophages and selectively promotes IL-1β transcription.","evidence":"siRNA silencing in primary human macrophages, cytokine qPCR, CAGE promoter analysis, human/mouse comparison","pmids":["26819317"],"confidence":"Medium","gaps":["molecular target mediating IL-1β induction not identified","ligase activity not connected to this phenotype"]},{"year":2018,"claim":"Showed RNF144B protein is stabilized by GSK3β-dependent phosphorylation, coupling its abundance to proliferative signaling in endometrial cancer.","evidence":"GSK3β inhibitor treatment, phosphorylation analysis, siRNA knockdown, proliferation assays","pmids":["29724995"],"confidence":"Medium","gaps":["specific phosphorylation site not mapped","direct vs indirect role of GSK3β not established"]},{"year":2019,"claim":"Identified RNF144B as a negative regulator of TBK1-IRF3 signaling, binding TBK1 via its IBR domain to suppress TBK1 phosphorylation, K63-ubiquitination, and IFN-β production.","evidence":"Co-IP with domain mapping (IBR-SDD), siRNA knockdown, Western blot for phospho/ubiquitination status, IFN-β ELISA","pmids":["31509299"],"confidence":"Medium","gaps":["whether ligase activity is required for TBK1 inhibition unclear","in vivo confirmation absent at this stage"]},{"year":2022,"claim":"Linked RNF144B to spermatogonial stem cell maintenance through an FCER2/NOTCH2/HES1 axis governing proliferation and survival.","evidence":"RNA-seq, Co-IP (RNF144B-FCER2, FCER2-N2ICD), siRNA knockdown, FCER2 overexpression rescue, proliferation/apoptosis assays","pmids":["35699595"],"confidence":"Medium","gaps":["whether FCER2 is a ubiquitination substrate not tested","ligase-dependence of the NOTCH2 effect unknown"]},{"year":2023,"claim":"Provided in vivo evidence that RNF144B tunes macrophage immune signaling, with myeloid-specific loss impairing TBK1 activation while enhancing NF-κB and worsening sepsis outcomes.","evidence":"Myeloid conditional knockout mice, Co-IP (Rnf144b-TBK1), Western blot, cytokine measurement, in vivo sepsis model","pmids":["37088470"],"confidence":"Medium","gaps":["molecular basis for divergent TBK1 vs NF-κB effects not resolved","substrate driving NF-κB suppression not identified at this point"]},{"year":2023,"claim":"Showed in ovarian cancer that HDAC3 induces RNF144B, which degrades the p21/p53 complex to promote a pro-tumorigenic phenotype, reinforcing p21 as a key substrate.","evidence":"Co-IP (RNF144B/p21/p53), cycloheximide chase, transcriptomics, knockdown/overexpression, in vivo tumor model","pmids":["37992458"],"confidence":"Medium","gaps":["ubiquitin linkage on the p21/p53 complex not defined","apparent oncogenic role here contrasts with tumor-suppressor findings, context dependence unexplained"]},{"year":2024,"claim":"Defined the molecular mechanism of RNF144B antiviral restraint: K27/K33-linked ubiquitination of MDA5 at K23/K43 in the CARDs directs p62-mediated autophagic degradation, with knockout boosting interferon and antiviral protection.","evidence":"Co-IP, ubiquitination linkage/site mapping, Rnf144b knockout mice, EMCV challenge, IFN assays, autophagy analysis","pmids":["39285245"],"confidence":"High","gaps":["E2 partner driving K27/K33 linkages not specified","interplay with the TBK1 branch of the same pathway not integrated"]},{"year":2024,"claim":"Established RNF144B as a TP53-downstream tumor suppressor maintaining genomic stability, with loss causing chromosomal instability and mitotic defects.","evidence":"Knockout in human/mouse non-transformed and cancer cells, proteomics/transcriptomics, cell cycle and DNA damage assays, in vivo tumor models","pmids":["38685100"],"confidence":"Medium","gaps":["specific degradation substrates driving genomic stability not pinpointed","reconciliation with oncogenic context-dependent roles incomplete"]},{"year":2026,"claim":"Identified TRAF3 as a K48-linked ubiquitination substrate of RNF144B, showing degradation of TRAF3 restrains NF-κB/MAPK signaling and limits neuroinflammation after ischemic stroke.","evidence":"Co-IP, K48-linked ubiquitination assay, RNF144B knockout mice (MCAO), TRAF3 knockdown epistasis rescue, Western blot, immunofluorescence","pmids":["41903812"],"confidence":"High","gaps":["E2 partner and structural basis of TRAF3 recognition not defined","relationship between TRAF3 and TBK1 branches of RNF144B immune regulation not integrated"]},{"year":null,"claim":"How RNF144B's apparently opposing roles — tumor suppressor maintaining genomic stability versus context-dependent pro-proliferative ligase — are reconciled, and what determines its substrate selection across cell types, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["no unifying model for substrate choice across apoptotic, immune, and cell-cycle contexts","structural determinants of linkage-type specificity (K48 vs K27/K33) unknown","physiological hierarchy among Bax, p21, ΔNp63α, MDA5, TRAF3 substrates undefined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[1,2,10,12]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,3,10,12]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0]},{"term_id":"GO:0005739","term_label":"mitochondrion","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[0,2]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[4,5,8,10,12]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[1,3,10,12]},{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[11]}],"complexes":[],"partners":["BAX","TP53","CDKN1A","TP63","TBK1","IFIH1","TRAF3","FCER2"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q7Z419","full_name":"E3 ubiquitin-protein ligase RNF144B","aliases":["IBR domain-containing protein 2","RING finger protein 144B","p53-inducible RING finger protein"],"length_aa":303,"mass_kda":33.7,"function":"E3 ubiquitin-protein ligase which accepts ubiquitin from E2 ubiquitin-conjugating enzymes UBE2L3 and UBE2L6 in the form of a thioester and then directly transfers the ubiquitin to targeted substrates such as LCMT2, thereby promoting their degradation. Induces apoptosis via a p53/TP53-dependent but caspase-independent mechanism. Plays a crucial role in maintaining the genomic stability by controlling the degradation of multiple proteins involved in mitotic progression and DNA damage (PubMed:38685100). Regulates epithelial homeostasis by mediating degradation of CDKN1A and isoform 2 of TP63 (PubMed:23128396). Plays a regulatory role in innate immunity by negatively regulating IRF3 activation and IFN-beta production. Mechanistically, inhibits TBK1 phosphorylation and 'Lys-63'-linked polyubiquitination independently of its E3 ligase activity (PubMed:31509299). Alternatively, promotes 'Lys-27' and 'Lys-33'-linked ubiquitination of IFIH1/MDA5, promoting selective autophagic degradation of IFIH1/MDA5 to inhibit antiviral response (PubMed:39285245)","subcellular_location":"Mitochondrion membrane; Cytoplasm","url":"https://www.uniprot.org/uniprotkb/Q7Z419/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF144B","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF144B","total_profiled":1310},"omim":[{"mim_id":"618869","title":"RING FINGER PROTEIN 144B; RNF144B","url":"https://www.omim.org/entry/618869"},{"mim_id":"603273","title":"TUMOR PROTEIN p63; TP63","url":"https://www.omim.org/entry/603273"},{"mim_id":"116899","title":"CYCLIN-DEPENDENT KINASE INHIBITOR 1A; CDKN1A","url":"https://www.omim.org/entry/116899"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoli","reliability":"Approved"},{"location":"Mitochondria","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"skeletal muscle","ntpm":53.3}],"url":"https://www.proteinatlas.org/search/RNF144B"},"hgnc":{"alias_symbol":["bA528A10.3","P53RFP"],"prev_symbol":["IBRDC2"]},"alphafold":{"accession":"Q7Z419","domains":[{"cath_id":"3.30.40.10","chopping":"27-94","consensus_level":"medium","plddt":94.2496,"start":27,"end":94},{"cath_id":"-","chopping":"115-168","consensus_level":"medium","plddt":86.7446,"start":115,"end":168},{"cath_id":"-","chopping":"182-252","consensus_level":"medium","plddt":83.5923,"start":182,"end":252}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7Z419","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q7Z419-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q7Z419-F1-predicted_aligned_error_v6.png","plddt_mean":82.31},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF144B","jax_strain_url":"https://www.jax.org/strain/search?query=RNF144B"},"sequence":{"accession":"Q7Z419","fasta_url":"https://rest.uniprot.org/uniprotkb/Q7Z419.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q7Z419/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q7Z419"}},"corpus_meta":[{"pmid":"20300062","id":"PMC_20300062","title":"IBRDC2, an IBR-type E3 ubiquitin ligase, is a regulatory factor for Bax and apoptosis activation.","date":"2010","source":"The EMBO journal","url":"https://pubmed.ncbi.nlm.nih.gov/20300062","citation_count":74,"is_preprint":false},{"pmid":"12853982","id":"PMC_12853982","title":"p53RFP, a p53-inducible RING-finger protein, regulates the stability of p21WAF1.","date":"2003","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/12853982","citation_count":44,"is_preprint":false},{"pmid":"16427630","id":"PMC_16427630","title":"The p53-inducible E3 ubiquitin ligase p53RFP induces p53-dependent apoptosis.","date":"2006","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/16427630","citation_count":32,"is_preprint":false},{"pmid":"23128396","id":"PMC_23128396","title":"PIR2/Rnf144B regulates epithelial homeostasis by mediating degradation of p21WAF1 and p63.","date":"2012","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/23128396","citation_count":25,"is_preprint":false},{"pmid":"31509299","id":"PMC_31509299","title":"RNF144B inhibits LPS-induced inflammatory responses via binding TBK1.","date":"2019","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/31509299","citation_count":24,"is_preprint":false},{"pmid":"32096180","id":"PMC_32096180","title":"LINC00662 triggers malignant progression of chordoma by the activation of RNF144B via targeting miR-16-5p.","date":"2020","source":"European review for medical and pharmacological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/32096180","citation_count":24,"is_preprint":false},{"pmid":"26819317","id":"PMC_26819317","title":"The E3 ubiquitin ligase RNF144B is LPS-inducible in human, but not mouse, macrophages and promotes inducible IL-1β expression.","date":"2016","source":"Journal of leukocyte biology","url":"https://pubmed.ncbi.nlm.nih.gov/26819317","citation_count":17,"is_preprint":false},{"pmid":"35699595","id":"PMC_35699595","title":"RNF144B stimulates the proliferation and inhibits the apoptosis of human spermatogonial stem cells via the FCER2/NOTCH2/HES1 pathway and its abnormality is associated with azoospermia.","date":"2022","source":"Journal of cellular physiology","url":"https://pubmed.ncbi.nlm.nih.gov/35699595","citation_count":16,"is_preprint":false},{"pmid":"29724995","id":"PMC_29724995","title":"Pir2/Rnf144b is a potential endometrial cancer biomarker that promotes cell proliferation.","date":"2018","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/29724995","citation_count":15,"is_preprint":false},{"pmid":"39285245","id":"PMC_39285245","title":"RNF144B negatively regulates antiviral immunity by targeting MDA5 for autophagic degradation.","date":"2024","source":"EMBO reports","url":"https://pubmed.ncbi.nlm.nih.gov/39285245","citation_count":10,"is_preprint":false},{"pmid":"38685100","id":"PMC_38685100","title":"The TP53-activated E3 ligase RNF144B is a tumour suppressor that prevents genomic instability.","date":"2024","source":"Journal of experimental & clinical cancer research : CR","url":"https://pubmed.ncbi.nlm.nih.gov/38685100","citation_count":10,"is_preprint":false},{"pmid":"37088470","id":"PMC_37088470","title":"Rnf144b alleviates the inflammatory responses and cardiac dysfunction in sepsis.","date":"2023","source":"ESC heart failure","url":"https://pubmed.ncbi.nlm.nih.gov/37088470","citation_count":10,"is_preprint":false},{"pmid":"37992458","id":"PMC_37992458","title":"RNF144B-mediated p21 degradation regulated by HDAC3 contribute to enhancing ovarian cancer growth and metastasis.","date":"2023","source":"Tissue & cell","url":"https://pubmed.ncbi.nlm.nih.gov/37992458","citation_count":7,"is_preprint":false},{"pmid":"40756570","id":"PMC_40756570","title":"RNF144A and RNF144B: Important molecules for health.","date":"2025","source":"Open life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40756570","citation_count":2,"is_preprint":false},{"pmid":"39567703","id":"PMC_39567703","title":"Integrated analysis ceRNA network of autophagy-related gene RNF144B in steroid-induced necrosis of the femoral head.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39567703","citation_count":1,"is_preprint":false},{"pmid":"39948107","id":"PMC_39948107","title":"Characterization of RNF144B and PPP2R2A identified by a novel approach using TCGA data in ovarian cancer.","date":"2025","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39948107","citation_count":0,"is_preprint":false},{"pmid":"41903812","id":"PMC_41903812","title":"RNF144B deficiency aggravated neuroinflammatory response induced by cerebral ischemic stroke via regulating TRAF3 ubiquitination.","date":"2026","source":"Biochemical pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/41903812","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.03.673891","title":"Single-cell transcriptomics and machine learning reveal RNF144B and C5AR1 as immune-related biomarkers and therapeutic targets in myocardial infarction","date":"2025-09-06","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.03.673891","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2024.10.10.24315203","title":"Genome-wide association study in chondrocalcinosis reveals ENPP1 as a candidate therapeutic target in calcium pyrophosphate deposition disease","date":"2024-10-11","source":"bioRxiv","url":"https://doi.org/10.1101/2024.10.10.24315203","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11979,"output_tokens":4010,"usd":0.048044,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11810,"output_tokens":4399,"usd":0.084513,"stage2_stop_reason":"end_turn"},"total_usd":0.132557,"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\": 2010,\n      \"finding\": \"IBRDC2 (RNF144B) is an IBR-type RING-finger E3 ubiquitin ligase that regulates Bax stability through ubiquitination-dependent degradation, protecting cells from spontaneous apoptosis. Upon apoptosis induction, IBRDC2 translocates from cytosol to mitochondrial domains enriched with active Bax, physically interacts with activated Bax, and this translocation requires both mitochondrial localization and apoptotic activation of Bax, and depends on Bcl-xL expression levels.\",\n      \"method\": \"Co-immunoprecipitation (physical interaction with activated Bax), active Bax-specific antibody staining, Bax mutant analysis in HCT116 Bax(-/-) cells, ubiquitination assays with IBRDC2 overexpression/knockdown, subcellular fractionation/live imaging\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, Bax mutant epistasis in defined Bax-null cells, multiple orthogonal methods (ubiquitination assay, fractionation, active Bax antibody), single rigorous study with robust controls\",\n      \"pmids\": [\"20300062\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"p53RFP (RNF144B) is a transcriptional target of p53 with E3 ubiquitin ligase activity that interacts with and ubiquitinates p21WAF1, promoting its degradation. Its expression is negatively correlated with p21WAF1 protein levels, suggesting it regulates p21WAF1 stability.\",\n      \"method\": \"E3 ubiquitin ligase activity assay, co-immunoprecipitation (p53RFP–p21WAF1 interaction), expression correlation analysis, transcriptional reporter assay for p53 induction\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — E3 ligase activity demonstrated in vitro, Co-IP interaction shown, single lab with two orthogonal methods\",\n      \"pmids\": [\"12853982\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"p53RFP (RNF144B) induces p53-dependent but caspase-independent apoptosis. Its RING-IBR-RING domain mediates interaction with E2 ubiquitin-conjugating enzymes UbcH7 and UbcH8 but not UbcH5. The conserved C-terminal domain (not the E3 ligase domain) is required and sufficient for apoptosis induction, indicating apoptosis does not require E3 ubiquitin ligase activity.\",\n      \"method\": \"Co-immunoprecipitation (E2 enzyme binding), domain deletion mutant analysis, apoptosis assays (caspase-independent), overexpression in cell lines\",\n      \"journal\": \"FEBS letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain mutant analysis combined with Co-IP for E2 binding specificity, single lab with two orthogonal methods\",\n      \"pmids\": [\"16427630\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"PIR2/RNF144B is a direct transcriptional target of ΔNp63α in keratinocytes. It binds and mediates proteasomal degradation of both p21WAF1/CIP1 and ΔNp63α itself, forming an auto-regulatory feedback loop. Depletion of PIR2/RNF144B impairs keratinocyte proliferation and differentiation with accumulation of p21WAF1/CIP1.\",\n      \"method\": \"Transcriptional reporter assay (ΔNp63α target), co-immunoprecipitation (PIR2–ΔNp63α binding), proteasome inhibitor rescue, siRNA knockdown with proliferation/differentiation phenotypic readout\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal binding shown by Co-IP, proteasomal degradation confirmed with inhibitor, loss-of-function phenotype with defined molecular readout, multiple orthogonal methods in one study\",\n      \"pmids\": [\"23128396\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"RNF144B interacts with TBK1 through its IBR domain binding the scaffold/dimerization domain (SDD) of TBK1, inhibiting TBK1 phosphorylation and K63-linked polyubiquitination, which leads to TBK1 inactivation, IRF3 dephosphorylation, and reduced IFN-β production in response to LPS stimulation. RNF144B knockdown increases IRF3 activation and IFN-β production.\",\n      \"method\": \"Co-immunoprecipitation (RNF144B–TBK1 interaction), domain mapping (IBR domain–SDD interaction), siRNA knockdown, Western blot for phosphorylation and ubiquitination status, IFN-β ELISA\",\n      \"journal\": \"Journal of leukocyte biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP with domain mapping, siRNA knockdown with defined signaling readouts, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"31509299\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"RNF144B is necessary for priming of inflammasome responses in primary human macrophages; specifically, it promotes LPS-inducible IL-1β mRNA expression but does not regulate several other LPS-inducible cytokines (e.g., IL-10, IFN-γ) or inflammasome components (procaspase-1, pro-IL-18). RNF144B is LPS-inducible in human but not mouse macrophages due to differences in transcription factor binding sites in the promoter.\",\n      \"method\": \"Gene silencing (siRNA) in primary human macrophages, cytokine mRNA quantification (qPCR), cap analysis of gene expression (CAGE) for promoter activity, comparative analysis across human and mouse cell types\",\n      \"journal\": \"Journal of leukocyte biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — gene silencing with specific cytokine readouts, CAGE promoter analysis, multiple cell types compared; single lab\",\n      \"pmids\": [\"26819317\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"PIR2/RNF144B protein is stabilized via phosphorylation downstream of GSK3β activity, and this stabilization is necessary for proliferation of endometrial cancer cells in the absence of oestrogenic growth stimuli. Inactivation of GSK3β leads to loss of PIR2/RNF144B protein and consequent inhibition of cell proliferation.\",\n      \"method\": \"GSK3β inhibitor treatment, phosphorylation analysis, siRNA knockdown, cell proliferation assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — pharmacological GSK3β inhibition linked to RNF144B stability loss and proliferation; single lab with two orthogonal methods but no direct demonstration of phosphorylation site\",\n      \"pmids\": [\"29724995\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF144B interacts with FCER2 in human spermatogonial stem cells (SSCs), and through FCER2 activates the NOTCH2/HES1 pathway (FCER2 pulls down NOTCH2 intracellular domain N2ICD). RNF144B knockdown reduces NOTCH2, FCER2, HES1, and HEY1 levels, inhibits SSC proliferation, and promotes apoptosis; these effects are reversed by FCER2 overexpression.\",\n      \"method\": \"RNA sequencing (target identification), co-immunoprecipitation (RNF144B–FCER2 and FCER2–N2ICD interactions), siRNA knockdown, FCER2 overexpression rescue, proliferation and apoptosis assays\",\n      \"journal\": \"Journal of cellular physiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP interactions, epistasis rescue experiment with FCER2 overexpression, RNA-seq target identification; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"35699595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF144B interacts with TBK1; in sepsis model, Rnf144b deficiency results in impaired TBK1 activation but enhanced NF-κB activation in macrophages. Conditional knockout of Rnf144b in myeloid cells leads to increased inflammatory cytokines after LPS/CpG stimulation, higher mortality, and exacerbated cardiac dysfunction in septic mice.\",\n      \"method\": \"Conditional knockout mice (myeloid-specific), co-immunoprecipitation (Rnf144b–TBK1), Western blot for TBK1 and NF-κB activation, cytokine measurement, in vivo sepsis model\",\n      \"journal\": \"ESC heart failure\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — conditional KO mice with defined in vivo phenotype, Co-IP, signaling readouts; single lab\",\n      \"pmids\": [\"37088470\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"HDAC3 promotes RNF144B expression in ovarian cancer cells. RNF144B interacts with p21 and regulates degradation of the p21/p53 complex. RNF144B-mediated p21 degradation (assessed by cycloheximide chase) promotes ovarian cancer cell proliferation, migration, and invasion while inhibiting apoptosis.\",\n      \"method\": \"Co-immunoprecipitation (RNF144B/p21/p53 interaction), cycloheximide chase (p21 stability), transcriptome profiling, siRNA knockdown and overexpression, in vivo tumor model\",\n      \"journal\": \"Tissue & cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP interaction, CHX chase for substrate stability, in vivo validation; single lab\",\n      \"pmids\": [\"37992458\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF144B specifically interacts with MDA5 and promotes K27/K33-linked polyubiquitination of MDA5 at lysine residues K23 and K43 (within the CARDs domain), which promotes autophagic degradation of MDA5 by p62. Rnf144b knockout in mice greatly promotes IFN production, inhibits EMCV replication, and significantly increases overall survival upon EMCV infection.\",\n      \"method\": \"Co-immunoprecipitation (RNF144B–MDA5 interaction), ubiquitination assay with linkage type and site identification (K27/K33, K23/K43), Rnf144b knockout mice, viral challenge (EMCV), IFN production assay, autophagy pathway analysis\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — ubiquitination site mapping, linkage-type determination, KO mouse in vivo phenotype with viral infection, multiple orthogonal methods including site-specific mutagenesis implied by lysine identification\",\n      \"pmids\": [\"39285245\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF144B acts as a tumor suppressor downstream of TP53 in lung adenocarcinoma, mediating protein degradation associated with cell cycle progression, DNA damage response, and genomic stability. RNF144B deficiency induces chromosomal instability, mitotic defects, and resistance to cell cycle inhibitors that induce chromosomal instability.\",\n      \"method\": \"RNF144B knockout in human and mouse cells (non-transformed and cancerous), proteomics and transcriptomics analysis, proliferation/transformation assays, cell cycle analysis, DNA damage response assays, in vivo tumor models\",\n      \"journal\": \"Journal of experimental & clinical cancer research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO models in multiple cell types, proteomics and transcriptomics with functional validation; single lab\",\n      \"pmids\": [\"38685100\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"RNF144B interacts with TRAF3 and promotes its K48-linked ubiquitination and proteasomal degradation. In the absence of RNF144B (KO mice), TRAF3 is stabilized, leading to enhanced NF-κB and MAPK signaling pathway activation and exacerbated neuroinflammation after ischemic stroke. TRAF3 knockdown in RNF144B-deficient mice partially reversed neurological dysfunction and neuroinflammation post-MCAO.\",\n      \"method\": \"Co-immunoprecipitation (RNF144B–TRAF3 interaction), ubiquitination assay (K48-linked), RNF144B knockout mice (MCAO model), TRAF3 knockdown epistasis rescue, Western blot for NF-κB and MAPK activation, immunofluorescence\",\n      \"journal\": \"Biochemical pharmacology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — Co-IP, K48-linked ubiquitination demonstrated, KO mouse in vivo phenotype, epistasis rescue with TRAF3 knockdown, multiple orthogonal methods\",\n      \"pmids\": [\"41903812\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF144B (also known as IBRDC2/p53RFP/PIR2) is a RING-IBR-RING domain E3 ubiquitin ligase that is transcriptionally induced by p53 and ΔNp63α, and acts as a multi-substrate ubiquitin ligase that targets p21WAF1, ΔNp63α, Bax, MDA5, and TRAF3 for ubiquitination-dependent degradation; it interacts with E2 enzymes UbcH7/UbcH8 via its RING-IBR-RING domain, is stabilized by GSK3β-dependent phosphorylation, translocates to mitochondria upon apoptosis to interact with activated Bax, suppresses innate immune signaling by binding TBK1 (via its IBR domain) to inhibit TBK1 phosphorylation/K63-ubiquitination and by promoting K27/K33-ubiquitination of MDA5 at K23/K43 to direct its autophagic degradation via p62, degrades TRAF3 via K48-linked ubiquitination to restrain NF-κB/MAPK signaling, and functions as a tumor suppressor maintaining genomic stability downstream of TP53.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RNF144B is a RING-IBR-RING (RBR) E3 ubiquitin ligase that operates downstream of the p53/p63 transcription factor network to control cell fate and innate immune signaling [#1, #11]. It is a direct transcriptional target of p53 and of ΔNp63α, and its RING-IBR-RING domain engages the E2 conjugating enzymes UbcH7 and UbcH8 to drive substrate ubiquitination [#1, #2, #3]. As a multi-substrate ligase, RNF144B promotes degradation of the cyclin-dependent kinase inhibitor p21WAF1/CIP1 and of ΔNp63α itself, the latter forming an auto-regulatory feedback loop required for normal keratinocyte proliferation and differentiation [#1, #3]. In apoptosis, RNF144B ubiquitinates Bax to restrain spontaneous cell death, and upon apoptotic stimulation it translocates from the cytosol to mitochondrial domains enriched in activated Bax [#0]. RNF144B is a recurrent negative regulator of innate immune signaling: it binds TBK1 through its IBR domain to suppress TBK1 phosphorylation, K63-ubiquitination, and downstream IRF3/IFN-β activation; it directs K27/K33-linked ubiquitination of MDA5 at K23/K43 within the CARDs to route MDA5 for p62-dependent autophagic degradation, limiting antiviral interferon responses; and it drives K48-linked ubiquitination and proteasomal degradation of TRAF3 to restrain NF-κB and MAPK signaling [#4, #10, #12]. Acting downstream of TP53, RNF144B functions as a tumor suppressor that maintains genomic stability, with its loss producing chromosomal instability and mitotic defects [#11].\",\n  \"teleology\": [\n    {\n      \"year\": 2003,\n      \"claim\": \"Established RNF144B as a p53-inducible E3 ligase, linking it to the tumor suppressor transcriptional program and identifying p21WAF1 as a degradation substrate.\",\n      \"evidence\": \"In vitro E3 ligase assay, Co-IP of p53RFP-p21WAF1, p53 transcriptional reporter\",\n      \"pmids\": [\"12853982\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"E2 partner and ubiquitin linkage type not defined\", \"in vivo relevance of p21 turnover untested\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Mapped E2 enzyme specificity (UbcH7/UbcH8, not UbcH5) to the RING-IBR-RING domain and showed RNF144B-induced apoptosis is caspase-independent and separable from its ligase activity.\",\n      \"evidence\": \"Co-IP for E2 binding, domain deletion mutants, apoptosis assays in cell lines\",\n      \"pmids\": [\"16427630\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"substrate ubiquitinated during apoptosis not identified\", \"mechanism of caspase-independent death by the C-terminal domain unresolved\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined RNF144B as a regulator of Bax stability that protects against spontaneous apoptosis and relocalizes to mitochondria to engage activated Bax during cell death.\",\n      \"evidence\": \"Reciprocal Co-IP with activated Bax, Bax mutant epistasis in HCT116 Bax-null cells, ubiquitination assays, subcellular fractionation/live imaging\",\n      \"pmids\": [\"20300062\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ubiquitin linkage type on Bax not defined\", \"trigger for cytosol-to-mitochondria translocation not mechanistically resolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Placed RNF144B in a ΔNp63α auto-regulatory feedback loop in keratinocytes, showing it degrades both p21WAF1 and ΔNp63α to enable proliferation and differentiation.\",\n      \"evidence\": \"ΔNp63α transcriptional reporter, reciprocal Co-IP, proteasome inhibitor rescue, siRNA knockdown with proliferation/differentiation readout\",\n      \"pmids\": [\"23128396\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"ubiquitin linkage on ΔNp63α not characterized\", \"in vivo epidermal phenotype untested\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Implicated RNF144B in inflammasome priming, showing it is LPS-inducible in human (not mouse) macrophages and selectively promotes IL-1β transcription.\",\n      \"evidence\": \"siRNA silencing in primary human macrophages, cytokine qPCR, CAGE promoter analysis, human/mouse comparison\",\n      \"pmids\": [\"26819317\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"molecular target mediating IL-1β induction not identified\", \"ligase activity not connected to this phenotype\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showed RNF144B protein is stabilized by GSK3β-dependent phosphorylation, coupling its abundance to proliferative signaling in endometrial cancer.\",\n      \"evidence\": \"GSK3β inhibitor treatment, phosphorylation analysis, siRNA knockdown, proliferation assays\",\n      \"pmids\": [\"29724995\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"specific phosphorylation site not mapped\", \"direct vs indirect role of GSK3β not established\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Identified RNF144B as a negative regulator of TBK1-IRF3 signaling, binding TBK1 via its IBR domain to suppress TBK1 phosphorylation, K63-ubiquitination, and IFN-β production.\",\n      \"evidence\": \"Co-IP with domain mapping (IBR-SDD), siRNA knockdown, Western blot for phospho/ubiquitination status, IFN-β ELISA\",\n      \"pmids\": [\"31509299\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"whether ligase activity is required for TBK1 inhibition unclear\", \"in vivo confirmation absent at this stage\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Linked RNF144B to spermatogonial stem cell maintenance through an FCER2/NOTCH2/HES1 axis governing proliferation and survival.\",\n      \"evidence\": \"RNA-seq, Co-IP (RNF144B-FCER2, FCER2-N2ICD), siRNA knockdown, FCER2 overexpression rescue, proliferation/apoptosis assays\",\n      \"pmids\": [\"35699595\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"whether FCER2 is a ubiquitination substrate not tested\", \"ligase-dependence of the NOTCH2 effect unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Provided in vivo evidence that RNF144B tunes macrophage immune signaling, with myeloid-specific loss impairing TBK1 activation while enhancing NF-κB and worsening sepsis outcomes.\",\n      \"evidence\": \"Myeloid conditional knockout mice, Co-IP (Rnf144b-TBK1), Western blot, cytokine measurement, in vivo sepsis model\",\n      \"pmids\": [\"37088470\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"molecular basis for divergent TBK1 vs NF-κB effects not resolved\", \"substrate driving NF-κB suppression not identified at this point\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showed in ovarian cancer that HDAC3 induces RNF144B, which degrades the p21/p53 complex to promote a pro-tumorigenic phenotype, reinforcing p21 as a key substrate.\",\n      \"evidence\": \"Co-IP (RNF144B/p21/p53), cycloheximide chase, transcriptomics, knockdown/overexpression, in vivo tumor model\",\n      \"pmids\": [\"37992458\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"ubiquitin linkage on the p21/p53 complex not defined\", \"apparent oncogenic role here contrasts with tumor-suppressor findings, context dependence unexplained\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined the molecular mechanism of RNF144B antiviral restraint: K27/K33-linked ubiquitination of MDA5 at K23/K43 in the CARDs directs p62-mediated autophagic degradation, with knockout boosting interferon and antiviral protection.\",\n      \"evidence\": \"Co-IP, ubiquitination linkage/site mapping, Rnf144b knockout mice, EMCV challenge, IFN assays, autophagy analysis\",\n      \"pmids\": [\"39285245\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"E2 partner driving K27/K33 linkages not specified\", \"interplay with the TBK1 branch of the same pathway not integrated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established RNF144B as a TP53-downstream tumor suppressor maintaining genomic stability, with loss causing chromosomal instability and mitotic defects.\",\n      \"evidence\": \"Knockout in human/mouse non-transformed and cancer cells, proteomics/transcriptomics, cell cycle and DNA damage assays, in vivo tumor models\",\n      \"pmids\": [\"38685100\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"specific degradation substrates driving genomic stability not pinpointed\", \"reconciliation with oncogenic context-dependent roles incomplete\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Identified TRAF3 as a K48-linked ubiquitination substrate of RNF144B, showing degradation of TRAF3 restrains NF-κB/MAPK signaling and limits neuroinflammation after ischemic stroke.\",\n      \"evidence\": \"Co-IP, K48-linked ubiquitination assay, RNF144B knockout mice (MCAO), TRAF3 knockdown epistasis rescue, Western blot, immunofluorescence\",\n      \"pmids\": [\"41903812\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"E2 partner and structural basis of TRAF3 recognition not defined\", \"relationship between TRAF3 and TBK1 branches of RNF144B immune regulation not integrated\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RNF144B's apparently opposing roles — tumor suppressor maintaining genomic stability versus context-dependent pro-proliferative ligase — are reconciled, and what determines its substrate selection across cell types, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"no unifying model for substrate choice across apoptotic, immune, and cell-cycle contexts\", \"structural determinants of linkage-type specificity (K48 vs K27/K33) unknown\", \"physiological hierarchy among Bax, p21, ΔNp63α, MDA5, TRAF3 substrates undefined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [1, 2, 10, 12]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 3, 10, 12]},\n      {\"term_id\": \"GO:0061630\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005739\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [4, 5, 8, 10, 12]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [1, 3, 10, 12]},\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"BAX\", \"TP53\", \"CDKN1A\", \"TP63\", \"TBK1\", \"IFIH1\", \"TRAF3\", \"FCER2\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}