{"gene":"RNF213","run_date":"2026-06-10T06:43:37","timeline":{"discoveries":[{"year":2011,"finding":"RNF213 encodes a 591-kDa cytosolic protein possessing a Walker motif (ATPase) and a RING finger domain that exhibit ATPase and ubiquitin ligase activities, respectively. Knockdown of RNF213 in zebrafish caused irregular wall formation in trunk arteries and abnormal sprouting vessels, establishing a role in vascular development.","method":"Exome sequencing/Sanger sequencing for variant identification; functional assays for ATPase/ubiquitin ligase activity; zebrafish morpholino knockdown with vascular phenotype readout","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — biochemical activity assays combined with in vivo zebrafish loss-of-function with defined vascular phenotype; foundational paper replicated across many subsequent studies","pmids":["21799892"],"is_preprint":false},{"year":2014,"finding":"Mysterin/RNF213 contains two tandem AAA+ ATPase modules and forms a large ring-shaped oligomeric complex. Fluorescence correlation spectroscopy and biochemical evaluation showed that RNF213 dynamically changes its oligomeric state through ATP/ADP binding and hydrolysis cycles, consistent with a mechanical AAA+ ATPase mechanism.","method":"Fluorescence correlation spectroscopy (FCS), biochemical sedimentation/native PAGE assays for oligomeric state, ATPase activity assays","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical reconstitution with multiple orthogonal methods (FCS + biochemical assays) in a single study","pmids":["24658080"],"is_preprint":false},{"year":2015,"finding":"RNF213 is upregulated by IFN-β through STAT signaling in endothelial cells and mediates antiangiogenic activity of IFN-β. The R4810K variant decreases ATPase activity (similar to a Walker B motif stabilizing mutation WEQ) and stabilizes oligomers, causing antiangiogenic activity even without IFN-β stimulation. EC-specific Rnf213 R4757K transgenic mice showed suppressed cerebral angiogenesis under hypoxia, while wild-type overexpression did not.","method":"Promoter analysis (STAT binding), ATPase activity assays, mutagenesis of Walker B motif, transgenic mouse hypoxia model with in vivo angiogenesis readout","journal":"Journal of the American Heart Association","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro biochemical assays with mutagenesis plus in vivo transgenic mouse model with functional angiogenesis readout","pmids":["26126547"],"is_preprint":false},{"year":2015,"finding":"Pro-inflammatory cytokines IFN-γ and TNF-α synergistically activate transcription of RNF213 in endothelial cells via AKT and PKR pathways. RNF213 knockdown in endothelial cells reduced expression of cell cycle-promoting genes, decreased proliferation, reduced angiogenic capacity, and down-regulated matrix metalloproteases specifically in endothelial cells.","method":"Chemical inhibitor experiments (LY294002 for AKT, C16 for PKR), RNAi knockdown, transcriptome-wide analysis, qPCR validation, proliferation and tube formation assays","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (transcriptomics + functional assays + inhibitor studies) in a single lab","pmids":["26278786"],"is_preprint":false},{"year":2015,"finding":"RNF213 R4810K variant downregulates Securin in iPSC-derived vascular endothelial cells, inhibiting angiogenic activity. Overexpression of R4810K (but not wild-type) RNF213 reduced tube formation and proliferation in HUVECs. siRNA knockdown of Securin phenocopied the angiogenic defect.","method":"iPSC differentiation to endothelial cells, gene expression profiling, overexpression of WT vs. R4810K in HUVECs, tube formation assay, siRNA knockdown of Securin","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (iPSC model + overexpression + siRNA) establishing substrate/pathway link","pmids":["23850618"],"is_preprint":false},{"year":2013,"finding":"RNF213 R4810K variant induces mitotic abnormalities: overexpression in HeLa cells extended mitosis 4-fold, and co-immunoprecipitation revealed that R4810K forms a complex with MAD2 more readily than wild-type RNF213. Fibroblasts and iPSC-derived endothelial cells from patients showed desynchronized MAD2 localization and increased aneuploidy.","method":"Live-cell imaging, co-immunoprecipitation, immunofluorescence, flow cytometry for aneuploidy, MAD2 depletion epistasis experiment","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP identifying RNF213/MAD2 complex plus functional mitotic phenotype with epistasis (MAD2 depletion)","pmids":["23994138"],"is_preprint":false},{"year":2016,"finding":"PTP1B negatively regulates RNF213 in HER2+ breast cancer cells. RNF213 knockdown reverses the effects of PTP1B deficiency on α-KG-dependent dioxygenase (α-KGDD) activity and non-mitochondrial oxygen consumption, establishing a PTP1B→RNF213→α-KGDD pathway required for tumor survival under hypoxia.","method":"RNF213 siRNA knockdown in PTP1B-deficient and wild-type HER2+ breast cancer cells, oxygen consumption measurements, α-KGDD activity assays, xenograft tumorigenicity rescue","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — epistasis established by knockdown rescue, multiple orthogonal functional readouts (NMOC, α-KGDD, xenograft growth), replicated across multiple cell lines","pmids":["27323329"],"is_preprint":false},{"year":2020,"finding":"Cryo-EM structure of mouse RNF213 (584 kDa) revealed an N-terminal stalk, a dynein-like core with six ATPase units, and a multidomain E3 module. Collaboration with UbcH7, a cysteine-reactive E2, indicates a RING-independent ubiquitin-transfer mechanism. Pathogenic MMD mutations cluster in the composite E3 domain.","method":"Cryo-EM structure determination, biochemical E2 collaboration assays with UbcH7, mutational mapping of disease variants onto structure","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 / Strong — cryo-EM structure with biochemical validation of ubiquitin transfer mechanism; landmark structural paper","pmids":["32573437"],"is_preprint":false},{"year":2020,"finding":"RING domain mutations from MMD patients reduce RNF213 ubiquitin ligase activity (using Ubc13/Uev1A as E2, generating K63-linked polyubiquitin chains in vitro). In full-length overexpression experiments in HEK293T cells, these mutations conversely enhanced NF-κB activation and caspase-3-mediated apoptosis in an AAA+ domain-dependent manner, suggesting NF-κB/apoptosis activities are negatively regulated by RNF213 E3 ligase activity.","method":"In vitro ubiquitination assays with purified RING domain, K63 linkage-specific antibodies, full-length overexpression in HEK293T, NF-κB reporter assay, caspase-3 activity assay, AAA+ domain deletion mutants","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 1 / Moderate — in vitro ubiquitination assay with linkage specificity plus cell-based functional studies; single lab","pmids":["32139119"],"is_preprint":false},{"year":2021,"finding":"RNF213 ubiquitylates the lipid A moiety of bacterial lipopolysaccharide (LPS) on cytosol-invading Salmonella, forming a ubiquitin coat. This requires the dynein-like core of RNF213 but not its RING domain; instead, an RZ finger mediates ubiquitylation. RNF213 is essential for recruitment of LUBAC (which adds M1-linked chains) and ubiquitin-dependent autophagy receptors, restricting Salmonella proliferation.","method":"Biochemical ubiquitylation assays identifying LPS as substrate, domain deletion/mutagenesis (RZ finger vs. RING domain), co-immunoprecipitation for LUBAC recruitment, bacterial colony-forming unit assays, autophagy receptor recruitment imaging","journal":"Nature","confidence":"High","confidence_rationale":"Tier 1 / Strong — reconstitution of LPS ubiquitylation with mechanistic domain mutagenesis, multiple functional readouts; landmark mechanistic paper","pmids":["34012115"],"is_preprint":false},{"year":2021,"finding":"UBC13 (UBE2N) is an E2 ubiquitin-conjugating enzyme for RNF213, identified by yeast two-hybrid screening with the RNF213 RING domain as bait, and confirmed by co-immunoprecipitation in vivo. RNF213 autoubiquitinates via K63-linked chains (not K48) in a UBC13-dependent manner. UBC13 knockdown and ubiquitination-dead RNF213 mutants reduced HUVEC cell migration and invasion, indicating the RNF213-UBC13 axis contributes to angiogenic activity.","method":"Yeast two-hybrid screening, co-immunoprecipitation in vivo, in vitro ubiquitination assay with K63/K48 linkage analysis, UBC13 siRNA knockdown, migration/invasion assays","journal":"FASEB bioAdvances","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — yeast two-hybrid plus Co-IP for binding, in vitro ubiquitination for mechanism, functional rescue experiments; single lab","pmids":["33842849"],"is_preprint":false},{"year":2022,"finding":"RNF213 uses E2-conjugating enzymes UBE2D2 and UBE2L3 for distinct ubiquitylation events: RNF213-UBE2D2 catalyzes K6 (and K48) poly-ubiquitylation in vitro, while RNF213-UBE2L3 catalyzes K6-, K11-, and K48-linked chains. MMD-associated SNPs encode proteins with decreased E3 activity. The most frequent MMD allele (p.R4810K) acts as a dominant-negative mutant that globally decreases ubiquitylation but does not affect ATPase activity.","method":"In vitro ubiquitination assays with purified components, ubiquitin linkage analysis, dominant-negative functional assay in cells, ATPase activity assays for MMD SNP alleles","journal":"Life science alliance","confidence":"High","confidence_rationale":"Tier 1 / Strong — rigorous in vitro reconstitution with multiple E2 enzymes, linkage specificity determined, mechanistic characterization of disease variants; multiple orthogonal methods","pmids":["35135845"],"is_preprint":false},{"year":2022,"finding":"RNF213 knockout in human cerebral endothelial cells (CRISPR-Cas9) caused morphological changes, increased blood-brain barrier permeability, downregulation and delocalization of PECAM-1, abnormal leukocyte transmigration, and secretion of proinflammatory cytokines, identifying RNF213 as a regulator of cerebral endothelium integrity.","method":"CRISPR-Cas9 knockout in hCMEC/D3 cells, permeability assays, immunofluorescence for junction proteins (PECAM-1), transendothelial leukocyte migration assay, cytokine ELISA","journal":"Stroke","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean CRISPR knockout with multiple orthogonal functional readouts; single lab","pmids":["34991336"],"is_preprint":false},{"year":2022,"finding":"IFN-γ-inducible RNF213 translocates to Toxoplasma gondii parasitophorous vacuoles (PVs) and facilitates PV ubiquitylation (linear and K63-linked) in human cells independently of LUBAC. RNF213 is required for IFN-γ-mediated growth restriction of T. gondii. The ATPase domain is required for RNF213 recruitment to the PVM, while loss of a critical histidine in the RZ finger abolishes ubiquitylation; both are needed for parasite growth restriction.","method":"CRISPR loss-of-function screen, domain mutagenesis (ATPase domain, RZ finger histidine), immunofluorescence for PV localization, ubiquitin linkage analysis, intracellular parasite growth assay","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 / Strong — domain mutagenesis establishing distinct roles of ATPase (recruitment) vs. RZ finger (ubiquitylation), functional growth restriction assay, confirmed independently across studies","pmids":["36154443"],"is_preprint":false},{"year":2022,"finding":"GarD, a Chlamydia trachomatis inclusion membrane protein, functions as a cis-acting stealth factor that bars IFN-γ-inducible RNF213 from targeting inclusions. In IFN-γ-primed human cells lacking garD, inclusions become decorated with linear ubiquitin in an RNF213-dependent manner and are destroyed, establishing RNF213 as an anti-Chlamydia E3 ligase.","method":"Genetic screen, garD loss-of-function mutants, RNF213 knockdown/knockout, immunofluorescence for ubiquitin on inclusions, bacterial growth assays","journal":"Cell host & microbe","confidence":"High","confidence_rationale":"Tier 2 / Strong — genetic screen plus functional validation with bacterial mutants and host RNF213 knockdown; mechanistic antagonism defined","pmids":["36084633"],"is_preprint":false},{"year":2023,"finding":"RNF213 directly interacts with the Replication and Transcription Activator (RTA) of KSHV and MHV-68, promotes K48-linked ubiquitination of RTA, and targets it for proteasome-dependent degradation, thereby inhibiting gammaherpesvirus de novo infection and lytic reactivation.","method":"Co-immunoprecipitation (RNF213-RTA interaction), in vitro ubiquitination assay (K48 linkage), proteasome inhibitor experiments, viral infection/reactivation assays with RNF213 overexpression and knockdown","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Moderate — direct binding (Co-IP) plus in vitro ubiquitination with linkage specificity plus proteasomal degradation mechanism plus functional viral restriction; multiple orthogonal methods in single study","pmids":["36917666"],"is_preprint":false},{"year":2023,"finding":"Genome-wide CRISPR screen identified RNF213 as necessary for IFN-γ-mediated restriction of T. gondii. The ATPase domain is required for RNF213 recruitment to the parasitophorous vacuole membrane (PVM), while the RZ finger domain is required for ubiquitination. Both are required for parasite growth restriction. RNF213-mediated restriction is independent of ATG5 (non-canonical autophagy). RNF213 overexpression in naive (non-IFN-γ stimulated) cells is sufficient to restrict T. gondii growth.","method":"Genome-wide CRISPR screen, targeted ISG CRISPR screen, ATPase and RZ finger domain mutational analysis, fluorescence imaging of PVM localization, intracellular growth assays, ATG5 knockout epistasis","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — genome-wide screen plus domain mutagenesis plus epistasis with ATG5; multiple orthogonal methods establishing mechanism","pmids":["38147552"],"is_preprint":false},{"year":2023,"finding":"RNF213 loss-of-function activates the Hippo pathway effector YAP/TAZ, promoting overexpression of VEGFR2 downstream. Inhibition of YAP/TAZ altered VEGFR2 trafficking from Golgi to plasma membrane and reversed RNF213 knockdown-induced pathological angiogenesis in endothelial cells and in vivo (RNF213-deficient mice and zebrafish).","method":"RNF213 knockdown in human brain microvascular endothelial cells, bulk RNA-seq and single-cell RNA-seq, YAP/TAZ inhibitor experiments, VEGFR2 trafficking assay (Golgi vs. plasma membrane fractionation), in vivo RNF213-KO mouse and zebrafish vascular phenotyping","journal":"Brain : a journal of neurology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pathway inhibitor epistasis plus transcriptomics plus in vivo validation; single lab","pmids":["37399508"],"is_preprint":false},{"year":2024,"finding":"RNF213 specifically promotes regulatory T (Treg) cell differentiation and attenuates autoimmune disease in an FOXO1-dependent manner. Mechanistically, RNF213 interacts with FOXO1 and promotes its nuclear translocation via K63-linked ubiquitination. RNF213 expression in CD4+ T cells is induced by IFN-β and is required for IFN-β therapeutic efficacy in multiple sclerosis models.","method":"Co-immunoprecipitation (RNF213-FOXO1 interaction), K63 ubiquitination assays, nuclear fractionation, Treg differentiation assays, autoimmune disease mouse models, IFN-β treatment experiments","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP establishing binding, ubiquitination assay with linkage specificity, nuclear translocation assay, functional Treg differentiation rescue; multiple orthogonal methods","pmids":["39013878"],"is_preprint":false},{"year":2019,"finding":"S-nitrosylation of RNF213 was detected in the P301S tauopathy mouse model cortex and hippocampus. S-nitrosylated RNF213 was associated with increased levels of NFAT-1 and FILAMIN-A, implicating RNF213 S-nitrosylation in activation of non-canonical Wnt/Ca²⁺ signaling in tauopathy.","method":"S-nitrosoproteome mass spectrometry, pathway analysis, immunoblotting for downstream effectors (pCaMKII, NFAT-1, FILAMIN-A) in transgenic mouse brain tissue","journal":"Translational psychiatry","confidence":"Low","confidence_rationale":"Tier 3 / Weak — proteomics identification of SNO-RNF213 with correlative downstream measurements; no direct functional validation of the modification on RNF213 activity","pmids":["30696811"],"is_preprint":false},{"year":2020,"finding":"Genetic ablation of mitochondrial matrix factors ClpP, Tfam, and Lonp1 potently induces Rnf213 transcript expression in multiple organs. Rnf213 is also induced by Poly(I:C)-triggered TLR3-mediated responses and IFN-γ treatment. This induction is only partially suppressed by PKR antagonist C16, suggesting RNF213 is induced by mitochondrial dysfunction/dsRNA-dependent inflammation.","method":"Rnf213 mRNA quantification in tissue from genetic KO mice (ClpP, Tfam, Lonp1), Poly(I:C) treatment, IFN-γ treatment, PKR inhibitor (C16) experiments","journal":"Neurogenetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple genetic KO models plus pharmacological inhibitor experiments establishing upstream induction pathway; single lab","pmids":["32342250"],"is_preprint":false},{"year":2014,"finding":"RNF213-deficient mice show suppressed vascular remodeling after common carotid artery ligation: the intima and medial layers were significantly thinner in RNF213-/- than wild-type mice after ligation. Under normal conditions, no spontaneous arterial abnormalities were observed up to 64 weeks, indicating RNF213 loss is insufficient alone to cause moyamoya disease.","method":"Cre-lox RNF213 knockout mouse generation, 9.4-T MRA, common carotid artery ligation model, Elastica-Masson staining of arterial wall","journal":"Brain research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse with defined vascular phenotype using quantitative histopathology and MRA; single lab","pmids":["24440776"],"is_preprint":false},{"year":2014,"finding":"RNF213-deficient mice showed enhanced angiogenesis and improved blood flow recovery after permanent femoral artery ligation compared to wild-type, indicating RNF213 functions to suppress angiogenesis under chronic ischemic conditions.","method":"RNF213-/- knockout mice, femoral artery ligation model, laser speckle flowmetry, vascular density quantification, ambulatory impairment scoring","journal":"Brain research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse with multiple functional readouts (blood flow, vascular density, behavioral); single lab","pmids":["25446450"],"is_preprint":false},{"year":2014,"finding":"RNF213-deficient mice showed significantly increased vascular MMP-9 expression at 1 and 7 days after common carotid artery ligation, and thinner vascular walls at 14 days, linking RNF213 loss to dysregulated MMP-9-dependent vascular remodeling.","method":"RNF213-/- knockout mice, common carotid artery ligation, immunohistochemistry/immunoblotting for MMP-9, Elastica-Masson staining","journal":"Neuroreport","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse with defined molecular and histological phenotype; single lab","pmids":["25383461"],"is_preprint":false},{"year":2015,"finding":"Mysterin/RNF213 is required for proper fast muscle formation and neuromuscular regulation in zebrafish. Morpholino-mediated knockdown caused significant reduction in fast myofibrils and immature projection of primary motoneurons, leading to severe motor deficits. Rescue with fast muscle-specific RNF213 expression reversed these phenotypes. Both AAA+ ATPase and ubiquitin ligase activities were indispensable for proper fast muscle formation.","method":"Zebrafish antisense morpholino knockdown, tissue-specific mRNA rescue, immunofluorescence for myofibrils and motoneuron projections, behavioral motor assays, ATPase-dead and RING-dead rescue constructs","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — loss-of-function with tissue-specific rescue, activity-dead mutagenesis establishing both enzymatic domains are necessary; multiple orthogonal readouts","pmids":["26530008"],"is_preprint":false},{"year":2018,"finding":"RNF213 is required for maintenance of cerebral blood flow under hypoperfusion. RNF213 knockout mice showed significantly more severe CBF reduction and impaired CBF restoration compared to wild-type after bilateral carotid stenosis. EC-specific Rnf213 mutant (R4810K ortholog) transgenic mice also showed impaired CBF restoration and reduced angiogenesis, establishing a vascular endothelial cell-autonomous role.","method":"Bilateral carotid artery stenosis surgery, arterial spin-labeling MRI for quantitative CBF measurements, histological angiogenesis analysis in RNF213 KO and EC-specific transgenic mice","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple mouse models (KO and EC-specific transgenic) with quantitative CBF and histological readouts; single lab","pmids":["29483617"],"is_preprint":false},{"year":2021,"finding":"RNF213 is required for resistance to Rift Valley fever virus in mice. CRISPR/Cas9 Rnf213-deficient C57BL/6 mice were significantly more susceptible to RVF than controls, with reduced average survival time post-infection.","method":"CRISPR/Cas9 Rnf213 knockout mouse, Rift Valley fever virus infection, survival analysis","journal":"Mammalian genome","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean CRISPR KO mouse with defined in vivo infectious disease phenotype; single lab","pmids":["33420513"],"is_preprint":false},{"year":2021,"finding":"RNF213 knockdown in macrophages and adipocytes reduces its expression, which is enhanced by pro-inflammatory TNF-α stimuli (LPS) and suppressed by PPARγ-mediated anti-inflammatory stimuli. PTP1B inactivation abolished RNF213 expression and enhanced adipogenesis through PPARγ. Constitutive RNF213 expression suppressed adipocyte differentiation by inhibiting PPARγ, establishing a TNF-α/PTP1B/RNF213/PPARγ pathway in adipogenesis.","method":"RNF213 knockdown in macrophages/adipocytes, LPS stimulation, PPARγ inhibitor/activator treatment, PTP1B inhibitor experiments, adipogenesis assays (Oil Red O staining), qPCR","journal":"Gene","confidence":"Low","confidence_rationale":"Tier 3 / Weak — predominantly transcriptional/expression-level readouts without direct biochemical demonstration of pathway hierarchy; single lab","pmids":["33333224"],"is_preprint":false},{"year":2021,"finding":"RNF213 knockdown in bone marrow-derived mesenchymal stem cells upregulates TGF-β1 at both protein and mRNA levels but does not affect VEGF expression, suggesting RNF213 normally suppresses TGF-β1 in these cells.","method":"Lentivirus-mediated shRNA knockdown of RNF213 in rat BMSCs, RT-qPCR, ELISA for TGF-β1 and VEGF expression","journal":"Experimental and therapeutic medicine","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single knockdown experiment with transcriptional/protein readout only; no mechanistic pathway established; single lab","pmids":["34373710"],"is_preprint":false},{"year":2022,"finding":"RNF213 knockdown in HUVECs disrupts angiogenesis partly through downregulation of DNA replication/proliferation pathways. Endothelial cells sensitized to LPS-induced inflammation after RNF213 KD showed retarded migration and enhanced macrophage transmigration. RNF213 KD also caused extensive changes in mRNA splicing. In vascular smooth muscle cells, RNF213 KD altered cytoskeletal organization and contractility.","method":"RNF213 siRNA knockdown in HUVECs and vSMCs, transcriptome sequencing, splicing analysis, angiogenesis assays, LPS stimulation, macrophage transmigration assays, co-culture models","journal":"Journal of cerebral blood flow and metabolism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — comprehensive transcriptome plus multiple functional assays across two cell types; single lab","pmids":["35754359"],"is_preprint":false},{"year":2023,"finding":"RNF213 knockout in mice causes significant pericyte reduction, blood-brain barrier impairment in cortex, microglia activation, elevated proinflammatory cytokines, and reduced tight junction proteins (Occludin, Claudin-5, ZO-1).","method":"Rnf213 knockout mice, immunofluorescence for pericyte markers and tight junction proteins, cytokine quantification, BBB permeability assays","journal":"Molecular neurobiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean KO mouse with multiple cellular phenotype readouts; single lab","pmids":["37438553"],"is_preprint":false},{"year":2020,"finding":"RNF213 R4810K variant-expressing HUVECs show autophagy inhibition (increased SQSTM1/p62 and LC3-II) and impaired endothelial function (tube formation) after oxygen-glucose deprivation (OGD). Rapamycin and cilostazol as autophagy inducers restored RNF213 R4810K cell function, linking R4810K to autophagic impairment under ischemic stress.","method":"Transfection of RNF213 WT vs. R4810K in HUVECs, OGD model, immunoblotting for autophagy markers (p62, LC3-II), tube formation assay, transmission electron microscopy for autophagic vesicles, rapamycin/cilostazol pharmacological rescue","journal":"Journal of cerebral blood flow and metabolism","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — WT vs. mutant comparison with pharmacological rescue and multiple readouts including EM; single lab","pmids":["38573771"],"is_preprint":false},{"year":2021,"finding":"RNF213 controls Listeria monocytogenes infection through regulation of DDAH1 transcription and production of nitric oxide (NO). RNF213 knockdown downregulates DDAH1, reducing NO production in macrophages from RNF213 KO mice, thereby impairing anti-Listeria defense.","method":"Mass spectrometry-based proteomics of RNF213-depleted cells, RT-qPCR validation, DDAH1 knockdown, NO production measurements, Listeria growth assays in RNF213 KO mouse macrophages","journal":"Frontiers in cellular and infection microbiology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — proteomics plus functional validation with KO mouse macrophages and pathway mechanism (DDAH1-NO axis); single lab","pmids":["34804992"],"is_preprint":false}],"current_model":"RNF213 is a 591 kDa cytosolic E3 ubiquitin ligase and AAA+ ATPase with a dynein-like six-unit ATPase core and a multidomain E3 module (including an RZ finger critical for ubiquitylation and a RING domain); it ubiquitylates non-proteinaceous substrates such as bacterial LPS via its RZ finger (RING-independently), ubiquitylates protein substrates (e.g., KSHV/MHV-68 RTA via K48 linkage for proteasomal degradation, FOXO1 via K63 linkage for nuclear translocation) through its RING domain in collaboration with multiple E2 enzymes (UBC13/UBE2N, UBE2D2, UBE2L3, UbcH7), is transcriptionally induced by IFN-γ, IFN-β, TNF-α, and cellular stress (mitochondrial dysfunction/TLR3 activation), and acts as an innate immune effector that restricts cytosolic bacteria (Salmonella, Listeria), parasites (Toxoplasma), and viruses (KSHV, MHV-68) by ubiquitin-coating pathogen-containing compartments; in vascular endothelial cells it regulates angiogenesis, blood-brain barrier integrity, and cerebral blood flow, and pathogenic MMD-associated variants (especially p.R4810K) act as dominant-negative alleles that globally impair ubiquitylation and destabilize RNF213 ATPase oligomerization, thereby dysregulating angiogenic and inflammatory signaling."},"narrative":{"mechanistic_narrative":"RNF213 is a giant (~591 kDa) cytosolic protein that couples a dynein-like AAA+ ATPase core to a multidomain E3 ubiquitin ligase module, functioning both as an innate immune effector and as a regulator of vascular homeostasis [PMID:21799892, PMID:32573437]. Its ATPase core forms a ring-shaped oligomer whose assembly is dynamically remodeled through ATP/ADP cycles [PMID:24658080], while its E3 module catalyzes ubiquitylation through two mechanistically distinct routes: a RING-independent RZ-finger mechanism that ubiquitylates the lipid A moiety of bacterial LPS, and a RING-dependent mechanism operating with multiple E2 enzymes (UBC13/UBE2N, UBE2D2, UBE2L3, UbcH7) to build diverse chain linkages [PMID:32573437, PMID:34012115, PMID:33842849, PMID:35135845]. As an interferon-inducible effector, RNF213 is recruited to pathogen-containing compartments—Salmonella, Toxoplasma parasitophorous vacuoles, and Chlamydia inclusions—where ATPase-dependent recruitment and RZ-finger-dependent ubiquitylation coat the surface in ubiquitin and recruit downstream LUBAC and autophagy machinery to restrict the pathogen [PMID:34012115, PMID:36154443, PMID:38147552, PMID:36084633]; it likewise targets viral proteins such as gammaherpesvirus RTA for K48-linked proteasomal degradation [PMID:36917666] and promotes Treg differentiation via K63-linked ubiquitylation and nuclear translocation of FOXO1 [PMID:39013878]. In vascular endothelium RNF213 regulates angiogenesis, cerebral blood flow, and blood-brain barrier integrity [PMID:25446450, PMID:29483617, PMID:37438553], acting in part through the UBC13 ubiquitylation axis [PMID:33842849] and through suppression of YAP/TAZ-VEGFR2 signaling [PMID:37399508]. Transcription of RNF213 is induced by IFN-β, IFN-γ, and TNF-α as well as by mitochondrial dysfunction and TLR3 activation [PMID:26126547, PMID:26278786, PMID:32342250]. The major moyamoya disease-associated allele p.R4810K behaves as a dominant-negative that globally impairs ubiquitylation and disrupts endothelial angiogenic function [PMID:35135845, PMID:26126547, PMID:23850618].","teleology":[{"year":2011,"claim":"Established RNF213 as a dual-activity enzyme and linked it to vascular development, framing the central question of how a single protein bridges enzymology and angiogenesis.","evidence":"Biochemical ATPase/ubiquitin ligase assays plus zebrafish morpholino knockdown with vascular readout","pmids":["21799892"],"confidence":"High","gaps":["No substrates identified","Mechanistic link between enzymatic activity and vascular phenotype undefined"]},{"year":2014,"claim":"Showed the AAA+ core assembles into a dynamic ring-shaped oligomer driven by ATP/ADP cycling, defining RNF213 as a mechanical AAA+ ATPase.","evidence":"Fluorescence correlation spectroscopy and biochemical oligomeric-state and ATPase assays","pmids":["24658080"],"confidence":"High","gaps":["Functional consequence of oligomeric remodeling unresolved","No structure available at this stage"]},{"year":2014,"claim":"In vivo mouse models established that RNF213 loss suppresses vascular remodeling and enhances ischemic angiogenesis while being insufficient alone to cause arterial disease, defining its role as a modulator rather than sole driver.","evidence":"RNF213 knockout mice in carotid ligation and femoral ligation models with MRA, flowmetry, and MMP-9 analysis","pmids":["24440776","25446450","25383461"],"confidence":"Medium","gaps":["Molecular substrate linking RNF213 to remodeling unidentified","MMP-9 regulation mechanism indirect"]},{"year":2015,"claim":"Connected RNF213 to interferon and inflammatory signaling, showing it mediates IFN-β antiangiogenic activity and is co-induced by IFN-γ/TNF-α, establishing it as a cytokine-responsive endothelial effector.","evidence":"Promoter/STAT analysis, AKT/PKR inhibitor studies, ATPase assays, transgenic mouse hypoxia angiogenesis model, RNAi with proliferation/tube formation readouts","pmids":["26126547","26278786","23850618"],"confidence":"High","gaps":["Direct ubiquitylation substrate in angiogenesis not defined","Securin link correlative"]},{"year":2016,"claim":"Placed RNF213 in a PTP1B-dependent metabolic pathway controlling α-KG-dependent dioxygenase activity and hypoxic tumor survival, extending its role beyond vasculature.","evidence":"siRNA knockdown rescue in HER2+ breast cancer cells, oxygen consumption and α-KGDD assays, xenograft tumorigenicity","pmids":["27323329"],"confidence":"High","gaps":["Direct enzymatic substrate in this pathway unknown","Mechanism of α-KGDD regulation indirect"]},{"year":2020,"claim":"The cryo-EM structure revealed the N-terminal stalk, six-unit dynein-like ATPase core, and composite E3 module, and uncovered a RING-independent, cysteine-reactive (UbcH7) ubiquitin-transfer mechanism while mapping disease mutations to the E3 region.","evidence":"Cryo-EM structure of mouse RNF213 with E2 collaboration assays and disease-variant mapping","pmids":["32573437"],"confidence":"High","gaps":["Substrate engagement geometry not resolved","Coupling between ATPase cycling and ubiquitin transfer not structurally defined"]},{"year":2020,"claim":"Defined the E2 repertoire and linkage outputs and demonstrated that disease alleles, particularly p.R4810K, act as dominant-negatives that globally impair ubiquitylation without altering ATPase activity, clarifying how variants cause loss of E3 function.","evidence":"In vitro ubiquitination with UBC13/Uev1A, UBE2D2, UBE2L3, linkage analysis, NF-κB and caspase reporters, dominant-negative cell assays, ATPase assays on disease alleles","pmids":["32139119","35135845"],"confidence":"High","gaps":["Physiological substrate(s) for each E2-linkage combination not all identified","How dominant-negative impairment maps to vascular pathology incomplete"]},{"year":2020,"claim":"Identified upstream inducers beyond classical cytokines, showing mitochondrial matrix dysfunction and TLR3/dsRNA signaling drive Rnf213 expression, broadening its stress-response regulation.","evidence":"Rnf213 mRNA quantification in ClpP/Tfam/Lonp1 KO mice, Poly(I:C), IFN-γ, and PKR inhibitor experiments","pmids":["32342250"],"confidence":"Medium","gaps":["Transcription factors mediating mitochondrial-stress induction undefined","Functional output of stress-induced RNF213 not established"]},{"year":2021,"claim":"Defined RNF213 as a direct innate immune effector that ubiquitylates non-proteinaceous LPS lipid A on cytosolic bacteria via its RZ finger (RING-independent), seeding LUBAC and autophagy recruitment to restrict pathogens.","evidence":"Biochemical ubiquitylation of LPS, RZ-finger vs RING domain mutagenesis, LUBAC Co-IP, autophagy receptor imaging, CFU assays on Salmonella; plus UBC13 yeast two-hybrid/Co-IP and autoubiquitylation analysis; plus Listeria DDAH1-NO axis and RVFV resistance models","pmids":["34012115","33842849","34804992","33420513"],"confidence":"High","gaps":["How RNF213 recognizes diverse pathogen surfaces unclear","Relationship between LPS ubiquitylation and protein-substrate ubiquitylation not unified"]},{"year":2022,"claim":"Generalized RNF213 pathogen restriction to eukaryotic parasites and other bacteria, showing ATPase-dependent recruitment and RZ-finger-dependent ubiquitylation of Toxoplasma vacuoles and Chlamydia inclusions, with pathogen stealth factors (GarD) antagonizing it.","evidence":"Genome-wide and ISG CRISPR screens, domain mutagenesis (ATPase recruitment vs RZ ubiquitylation), PVM/inclusion imaging, ATG5 epistasis, garD bacterial mutants, growth restriction assays","pmids":["36154443","38147552","36084633"],"confidence":"High","gaps":["Signal directing RNF213 to specific compartments unknown","Downstream effector requirements vary across pathogens"]},{"year":2023,"claim":"Extended RNF213's E3 function to protein substrates in immunity, showing K48-linked degradation of gammaherpesvirus RTA to restrict viral lytic reactivation.","evidence":"Co-IP, in vitro K48 ubiquitination, proteasome inhibitor experiments, viral infection/reactivation assays","pmids":["36917666"],"confidence":"High","gaps":["Whether RING vs RZ finger drives RTA ubiquitylation not delineated","E2 partner for this event unspecified"]},{"year":2023,"claim":"Linked endothelial RNF213 loss to pathological angiogenesis via YAP/TAZ-driven VEGFR2 overexpression and trafficking, and to blood-brain barrier integrity through pericyte and tight-junction regulation.","evidence":"RNF213 knockdown with RNA-seq/scRNA-seq, YAP/TAZ inhibitor epistasis, VEGFR2 trafficking fractionation, in vivo mouse/zebrafish phenotyping; plus Rnf213 KO mouse BBB/pericyte analysis","pmids":["37399508","37438553"],"confidence":"Medium","gaps":["Direct ubiquitylation substrate upstream of YAP/TAZ unidentified","Connection to immune effector function unclear"]},{"year":2024,"claim":"Defined a substrate-level immunoregulatory role, showing RNF213 promotes Treg differentiation by K63-ubiquitylating FOXO1 to drive its nuclear translocation and is required for IFN-β therapeutic efficacy.","evidence":"Co-IP, K63 ubiquitination assays, nuclear fractionation, Treg differentiation and autoimmune disease mouse models, IFN-β treatment","pmids":["39013878"],"confidence":"High","gaps":["Whether FOXO1 ubiquitylation uses RING or RZ finger not stated","Structural basis of K63 vs K48 substrate selection unresolved"]},{"year":null,"claim":"How RNF213's two enzymatic activities are mechanistically coupled, and how a single ligase integrates pathogen restriction, vascular homeostasis, and immunometabolic signaling through distinct substrates and linkage types, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying model connecting ATPase cycling to substrate-specific ubiquitylation","The full physiological substrate set is incomplete","Causal mechanism by which p.R4810K produces moyamoya pathology in vivo not established"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,7,9,10,11,15,18]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[9,15,18]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[0,1,7,13]},{"term_id":"GO:0016874","term_label":"ligase activity","supporting_discovery_ids":[0,7,11]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,7]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[9,13,14,15,16,18]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[9,11,15,18]},{"term_id":"R-HSA-1266738","term_label":"Developmental Biology","supporting_discovery_ids":[0,17,22,25]}],"complexes":[],"partners":["UBE2N","UBE2D2","UBE2L3","FOXO1","MAD2","PTP1B"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q63HN8","full_name":"E3 ubiquitin-protein ligase RNF213","aliases":["ALK lymphoma oligomerization partner on chromosome 17","E3 ubiquitin-lipopolysaccharide ligase RNF213","Mysterin","RING finger protein 213"],"length_aa":5207,"mass_kda":591.4,"function":"Atypical E3 ubiquitin ligase that can catalyze ubiquitination of both proteins and lipids, and which is involved in various processes, such as lipid metabolism, angiogenesis and cell-autonomous immunity (PubMed:21799892, PubMed:26126547, PubMed:26278786, PubMed:26766444, PubMed:30705059, PubMed:32139119, PubMed:34012115). Acts as a key immune sensor by catalyzing ubiquitination of the lipid A moiety of bacterial lipopolysaccharide (LPS) via its RZ-type zinc-finger: restricts the proliferation of cytosolic bacteria, such as Salmonella, by generating the bacterial ubiquitin coat through the ubiquitination of LPS (PubMed:34012115). Also acts indirectly by mediating the recruitment of the LUBAC complex, which conjugates linear polyubiquitin chains (PubMed:34012115). Ubiquitination of LPS triggers cell-autonomous immunity, such as antibacterial autophagy, leading to degradation of the microbial invader (PubMed:34012115). Involved in lipid metabolism by regulating fat storage and lipid droplet formation; act by inhibiting the lipolytic process (PubMed:30705059). Also regulates lipotoxicity by inhibiting desaturation of fatty acids (PubMed:30846318). Also acts as an E3 ubiquitin-protein ligase via its RING-type zinc finger: mediates 'Lys-63'-linked ubiquitination of target proteins (PubMed:32139119, PubMed:33842849). Involved in the non-canonical Wnt signaling pathway in vascular development: acts by mediating ubiquitination and degradation of FLNA and NFATC2 downstream of RSPO3, leading to inhibit the non-canonical Wnt signaling pathway and promoting vessel regression (PubMed:26766444). Also has ATPase activity; ATPase activity is required for ubiquitination of LPS (PubMed:34012115)","subcellular_location":"Cytoplasm, cytosol; Lipid droplet","url":"https://www.uniprot.org/uniprotkb/Q63HN8/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/RNF213","classification":"Not Classified","n_dependent_lines":1,"n_total_lines":1208,"dependency_fraction":0.0008278145695364238},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/RNF213","total_profiled":1310},"omim":[{"mim_id":"613768","title":"RING FINGER PROTEIN 213; RNF213","url":"https://www.omim.org/entry/613768"},{"mim_id":"607151","title":"MOYAMOYA DISEASE 2; MYMY2","url":"https://www.omim.org/entry/607151"},{"mim_id":"252350","title":"MOYAMOYA DISEASE 1; MYMY1","url":"https://www.omim.org/entry/252350"},{"mim_id":"105590","title":"ALK RECEPTOR TYROSINE KINASE; ALK","url":"https://www.omim.org/entry/105590"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Cytosol","reliability":"Supported"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/RNF213"},"hgnc":{"alias_symbol":["KIAA1554","NET57","ALO17"],"prev_symbol":["C17orf27","KIAA1618","MYMY2"]},"alphafold":{"accession":"Q63HN8","domains":[],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q63HN8","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q63HN8-6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q63HN8-6-F1-predicted_aligned_error_v6.png","plddt_mean":86.25},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=RNF213","jax_strain_url":"https://www.jax.org/strain/search?query=RNF213"},"sequence":{"accession":"Q63HN8","fasta_url":"https://rest.uniprot.org/uniprotkb/Q63HN8.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q63HN8/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q63HN8"}},"corpus_meta":[{"pmid":"21799892","id":"PMC_21799892","title":"Identification of RNF213 as a susceptibility gene for moyamoya disease and its possible role in vascular development.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21799892","citation_count":569,"is_preprint":false},{"pmid":"34012115","id":"PMC_34012115","title":"Ubiquitylation of lipopolysaccharide by RNF213 during bacterial infection.","date":"2021","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/34012115","citation_count":292,"is_preprint":false},{"pmid":"22377813","id":"PMC_22377813","title":"Homozygous c.14576G>A variant of RNF213 predicts early-onset and severe form of moyamoya disease.","date":"2012","source":"Neurology","url":"https://pubmed.ncbi.nlm.nih.gov/22377813","citation_count":249,"is_preprint":false},{"pmid":"24949311","id":"PMC_24949311","title":"Genetics and Biomarkers of Moyamoya Disease: Significance of RNF213 as a Susceptibility Gene.","date":"2014","source":"Journal of stroke","url":"https://pubmed.ncbi.nlm.nih.gov/24949311","citation_count":132,"is_preprint":false},{"pmid":"25278557","id":"PMC_25278557","title":"RNF213 rare variants in an ethnically diverse population with Moyamoya disease.","date":"2014","source":"Stroke","url":"https://pubmed.ncbi.nlm.nih.gov/25278557","citation_count":130,"is_preprint":false},{"pmid":"26126547","id":"PMC_26126547","title":"Biochemical and Functional Characterization of RNF213 (Mysterin) R4810K, a Susceptibility Mutation of Moyamoya Disease, in Angiogenesis In Vitro and In Vivo.","date":"2015","source":"Journal of the American Heart Association","url":"https://pubmed.ncbi.nlm.nih.gov/26126547","citation_count":128,"is_preprint":false},{"pmid":"26278786","id":"PMC_26278786","title":"Moyamoya disease susceptibility gene RNF213 links inflammatory and angiogenic signals in endothelial cells.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26278786","citation_count":118,"is_preprint":false},{"pmid":"23110205","id":"PMC_23110205","title":"Molecular analysis of RNF213 gene for moyamoya disease in the Chinese Han population.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23110205","citation_count":110,"is_preprint":false},{"pmid":"32573437","id":"PMC_32573437","title":"Moyamoya disease factor RNF213 is a giant E3 ligase with a dynein-like core and a distinct ubiquitin-transfer mechanism.","date":"2020","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/32573437","citation_count":110,"is_preprint":false},{"pmid":"27323329","id":"PMC_27323329","title":"PTP1B controls non-mitochondrial oxygen consumption by regulating RNF213 to promote tumour survival during hypoxia.","date":"2016","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/27323329","citation_count":107,"is_preprint":false},{"pmid":"23850618","id":"PMC_23850618","title":"Downregulation of Securin by the variant RNF213 R4810K (rs112735431, G>A) reduces angiogenic activity of induced pluripotent stem cell-derived vascular endothelial cells from moyamoya patients.","date":"2013","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/23850618","citation_count":107,"is_preprint":false},{"pmid":"26662949","id":"PMC_26662949","title":"A new horizon of moyamoya disease and associated health risks explored through RNF213.","date":"2015","source":"Environmental health and preventive medicine","url":"https://pubmed.ncbi.nlm.nih.gov/26662949","citation_count":103,"is_preprint":false},{"pmid":"28635953","id":"PMC_28635953","title":"Rare RNF213 variants in the C-terminal region encompassing the RING-finger domain are associated with moyamoya angiopathy in Caucasians.","date":"2017","source":"European journal of human genetics : EJHG","url":"https://pubmed.ncbi.nlm.nih.gov/28635953","citation_count":96,"is_preprint":false},{"pmid":"24440776","id":"PMC_24440776","title":"Temporal profile of the vascular anatomy evaluated by 9.4-T magnetic resonance angiography and histopathological analysis in mice lacking RNF213: a susceptibility gene for moyamoya disease.","date":"2014","source":"Brain research","url":"https://pubmed.ncbi.nlm.nih.gov/24440776","citation_count":93,"is_preprint":false},{"pmid":"24658080","id":"PMC_24658080","title":"Moyamoya disease-associated protein mysterin/RNF213 is a novel AAA+ ATPase, which dynamically changes its oligomeric state.","date":"2014","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/24658080","citation_count":92,"is_preprint":false},{"pmid":"31650369","id":"PMC_31650369","title":"Moyamoya Disease and Spectrums of RNF213 Vasculopathy.","date":"2019","source":"Translational stroke research","url":"https://pubmed.ncbi.nlm.nih.gov/31650369","citation_count":90,"is_preprint":false},{"pmid":"26430847","id":"PMC_26430847","title":"Importance of RNF213 polymorphism on clinical features and long-term outcome in moyamoya disease.","date":"2015","source":"Journal of neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/26430847","citation_count":85,"is_preprint":false},{"pmid":"27128593","id":"PMC_27128593","title":"RNF213 as the major susceptibility gene for Chinese patients with moyamoya disease and its clinical relevance.","date":"2016","source":"Journal of neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/27128593","citation_count":71,"is_preprint":false},{"pmid":"34381413","id":"PMC_34381413","title":"RNF213 and GUCY1A3 in Moyamoya Disease: Key Regulators of Metabolism, Inflammation, and Vascular Stability.","date":"2021","source":"Frontiers in neurology","url":"https://pubmed.ncbi.nlm.nih.gov/34381413","citation_count":70,"is_preprint":false},{"pmid":"31949090","id":"PMC_31949090","title":"Predictive role of heterozygous p.R4810K of RNF213 in the phenotype of Chinese moyamoya disease.","date":"2020","source":"Neurology","url":"https://pubmed.ncbi.nlm.nih.gov/31949090","citation_count":67,"is_preprint":false},{"pmid":"25446450","id":"PMC_25446450","title":"Enhanced post-ischemic angiogenesis in mice lacking RNF213; a susceptibility gene for moyamoya disease.","date":"2014","source":"Brain research","url":"https://pubmed.ncbi.nlm.nih.gov/25446450","citation_count":63,"is_preprint":false},{"pmid":"22878964","id":"PMC_22878964","title":"P.R4810K, a polymorphism of RNF213, the susceptibility gene for moyamoya disease, is associated with blood pressure.","date":"2012","source":"Environmental health and preventive medicine","url":"https://pubmed.ncbi.nlm.nih.gov/22878964","citation_count":61,"is_preprint":false},{"pmid":"34991336","id":"PMC_34991336","title":"Moyamoya Disease Susceptibility Gene RNF213 Regulates Endothelial Barrier Function.","date":"2022","source":"Stroke","url":"https://pubmed.ncbi.nlm.nih.gov/34991336","citation_count":53,"is_preprint":false},{"pmid":"37399508","id":"PMC_37399508","title":"RNF213 loss-of-function promotes pathological angiogenesis in moyamoya disease via the Hippo pathway.","date":"2023","source":"Brain : a journal of neurology","url":"https://pubmed.ncbi.nlm.nih.gov/37399508","citation_count":52,"is_preprint":false},{"pmid":"32139119","id":"PMC_32139119","title":"Moyamoya disease patient mutations in the RING domain of RNF213 reduce its ubiquitin ligase activity and enhance NFκB activation and apoptosis in an AAA+ domain-dependent manner.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/32139119","citation_count":52,"is_preprint":false},{"pmid":"36084633","id":"PMC_36084633","title":"The bacterial effector GarD shields Chlamydia trachomatis inclusions from RNF213-mediated ubiquitylation and destruction.","date":"2022","source":"Cell host & microbe","url":"https://pubmed.ncbi.nlm.nih.gov/36084633","citation_count":52,"is_preprint":false},{"pmid":"27745834","id":"PMC_27745834","title":"RNF213 Is Associated with Intracranial Aneurysms in the French-Canadian Population.","date":"2016","source":"American journal of human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/27745834","citation_count":49,"is_preprint":false},{"pmid":"26315378","id":"PMC_26315378","title":"Temporal profile of the vascular anatomy evaluated by 9.4-tesla magnetic resonance angiography and histological analysis in mice with the R4859K mutation of RNF213, the susceptibility gene for moyamoya disease.","date":"2015","source":"Brain research","url":"https://pubmed.ncbi.nlm.nih.gov/26315378","citation_count":48,"is_preprint":false},{"pmid":"23994138","id":"PMC_23994138","title":"The moyamoya disease susceptibility variant RNF213 R4810K (rs112735431) induces genomic instability by mitotic abnormality.","date":"2013","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/23994138","citation_count":48,"is_preprint":false},{"pmid":"29165161","id":"PMC_29165161","title":"Rare variants of RNF213 and moyamoya/non-moyamoya intracranial artery stenosis/occlusion disease risk: a meta-analysis and systematic review.","date":"2017","source":"Environmental health and preventive medicine","url":"https://pubmed.ncbi.nlm.nih.gov/29165161","citation_count":47,"is_preprint":false},{"pmid":"27253870","id":"PMC_27253870","title":"A Polymorphism in RNF213 Is a Susceptibility Gene for Intracranial Atherosclerosis.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/27253870","citation_count":47,"is_preprint":false},{"pmid":"31060437","id":"PMC_31060437","title":"Prevalence of RNF213 p.R4810K Variant in Early-Onset Stroke With Intracranial Arterial Stenosis.","date":"2019","source":"Stroke","url":"https://pubmed.ncbi.nlm.nih.gov/31060437","citation_count":46,"is_preprint":false},{"pmid":"23769926","id":"PMC_23769926","title":"Impacts and interactions of PDGFRB, MMP-3, TIMP-2, and RNF213 polymorphisms on the risk of Moyamoya disease in Han Chinese human subjects.","date":"2013","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/23769926","citation_count":46,"is_preprint":false},{"pmid":"30696811","id":"PMC_30696811","title":"S-nitrosylation of E3 ubiquitin-protein ligase RNF213 alters non-canonical Wnt/Ca+2 signaling in the P301S mouse model of tauopathy.","date":"2019","source":"Translational psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/30696811","citation_count":46,"is_preprint":false},{"pmid":"36154443","id":"PMC_36154443","title":"Interferon-Inducible E3 Ligase RNF213 Facilitates Host-Protective Linear and K63-Linked Ubiquitylation of Toxoplasma gondii Parasitophorous Vacuoles.","date":"2022","source":"mBio","url":"https://pubmed.ncbi.nlm.nih.gov/36154443","citation_count":41,"is_preprint":false},{"pmid":"29718794","id":"PMC_29718794","title":"Rare variants in RNF213, a susceptibility gene for moyamoya disease, are found in patients with pulmonary hypertension and aggravate hypoxia-induced pulmonary hypertension in mice.","date":"2018","source":"Pulmonary circulation","url":"https://pubmed.ncbi.nlm.nih.gov/29718794","citation_count":41,"is_preprint":false},{"pmid":"31542298","id":"PMC_31542298","title":"Poor outcomes in carriers of the RNF213 variant (p.Arg4810Lys) with pulmonary arterial hypertension.","date":"2019","source":"The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation","url":"https://pubmed.ncbi.nlm.nih.gov/31542298","citation_count":40,"is_preprint":false},{"pmid":"26877770","id":"PMC_26877770","title":"Whole genome sequencing of \"Faecalibaculum rodentium\" ALO17, isolated from C57BL/6J laboratory mouse feces.","date":"2016","source":"Gut pathogens","url":"https://pubmed.ncbi.nlm.nih.gov/26877770","citation_count":39,"is_preprint":false},{"pmid":"25383461","id":"PMC_25383461","title":"Increased vascular MMP-9 in mice lacking RNF213: moyamoya disease susceptibility gene.","date":"2014","source":"Neuroreport","url":"https://pubmed.ncbi.nlm.nih.gov/25383461","citation_count":39,"is_preprint":false},{"pmid":"35562882","id":"PMC_35562882","title":"Novel Multifaceted Roles for RNF213 Protein.","date":"2022","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/35562882","citation_count":38,"is_preprint":false},{"pmid":"23466837","id":"PMC_23466837","title":"RNF213 polymorphism and Moyamoya disease: A systematic review and meta-analysis.","date":"2013","source":"Neurology India","url":"https://pubmed.ncbi.nlm.nih.gov/23466837","citation_count":38,"is_preprint":false},{"pmid":"23410753","id":"PMC_23410753","title":"Ablation of Rnf213 retards progression of diabetes in the Akita mouse.","date":"2013","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/23410753","citation_count":38,"is_preprint":false},{"pmid":"29483617","id":"PMC_29483617","title":"Dysregulation of RNF213 promotes cerebral hypoperfusion.","date":"2018","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/29483617","citation_count":37,"is_preprint":false},{"pmid":"33333224","id":"PMC_33333224","title":"New insights into TNFα/PTP1B and PPARγ pathway through RNF213- a link between inflammation, obesity, insulin resistance, and Moyamoya disease.","date":"2020","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/33333224","citation_count":36,"is_preprint":false},{"pmid":"27736983","id":"PMC_27736983","title":"RNF213 Rare Variants in Slovakian and Czech Moyamoya Disease Patients.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/27736983","citation_count":35,"is_preprint":false},{"pmid":"33420513","id":"PMC_33420513","title":"The ring finger protein 213 gene (Rnf213) contributes to Rift Valley fever resistance in mice.","date":"2021","source":"Mammalian genome : official journal of the International Mammalian Genome Society","url":"https://pubmed.ncbi.nlm.nih.gov/33420513","citation_count":31,"is_preprint":false},{"pmid":"27476341","id":"PMC_27476341","title":"Significant Association of the RNF213 p.R4810K Polymorphism with Quasi-Moyamoya Disease.","date":"2016","source":"Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association","url":"https://pubmed.ncbi.nlm.nih.gov/27476341","citation_count":31,"is_preprint":false},{"pmid":"39013878","id":"PMC_39013878","title":"RNF213 promotes Treg cell differentiation by facilitating K63-linked ubiquitination and nuclear translocation of FOXO1.","date":"2024","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/39013878","citation_count":29,"is_preprint":false},{"pmid":"26972532","id":"PMC_26972532","title":"Temporal profile of magnetic resonance angiography and decreased ratio of regulatory T cells after immunological adjuvant administration to mice lacking RNF213, a susceptibility gene for moyamoya disease.","date":"2016","source":"Brain research","url":"https://pubmed.ncbi.nlm.nih.gov/26972532","citation_count":29,"is_preprint":false},{"pmid":"35455046","id":"PMC_35455046","title":"RNF213-Associated Vascular Disease: A Concept Unifying Various Vasculopathies.","date":"2022","source":"Life (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/35455046","citation_count":27,"is_preprint":false},{"pmid":"26849809","id":"PMC_26849809","title":"Association between moyamoya syndrome and the RNF213 c.14576G>A variant in patients with neurofibromatosis Type 1.","date":"2016","source":"Journal of neurosurgery. Pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/26849809","citation_count":27,"is_preprint":false},{"pmid":"38147552","id":"PMC_38147552","title":"Genome-wide and targeted CRISPR screens identify RNF213 as a mediator of interferon gamma-dependent pathogen restriction in human cells.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/38147552","citation_count":26,"is_preprint":false},{"pmid":"28659337","id":"PMC_28659337","title":"Anaplastic lymphoma kinase-positive anaplastic large cell lymphoma with the variant RNF213-, ATIC- and TPM3-ALK fusions is characterized by copy number gain of the rearranged ALK gene.","date":"2017","source":"Haematologica","url":"https://pubmed.ncbi.nlm.nih.gov/28659337","citation_count":26,"is_preprint":false},{"pmid":"25956231","id":"PMC_25956231","title":"Mutation genotypes of RNF213 gene from moyamoya patients in Taiwan.","date":"2015","source":"Journal of the neurological sciences","url":"https://pubmed.ncbi.nlm.nih.gov/25956231","citation_count":25,"is_preprint":false},{"pmid":"29752070","id":"PMC_29752070","title":"RNF213 p.R4810K Polymorphism and the Risk of Moyamoya Disease, Intracranial Major Artery Stenosis/Occlusion, and Quasi-Moyamoya Disease: A Meta-Analysis.","date":"2018","source":"Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association","url":"https://pubmed.ncbi.nlm.nih.gov/29752070","citation_count":25,"is_preprint":false},{"pmid":"30283986","id":"PMC_30283986","title":"Posterior circulation involvement and collateral flow pattern in moyamoya disease with the RNF213 polymorphism.","date":"2018","source":"Child's nervous system : ChNS : official journal of the International Society for Pediatric Neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/30283986","citation_count":24,"is_preprint":false},{"pmid":"28797616","id":"PMC_28797616","title":"Genetic Analysis of Ring Finger Protein 213 (RNF213) c.14576G>A in Intracranial Atherosclerosis of the Anterior and Posterior Circulations.","date":"2017","source":"Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association","url":"https://pubmed.ncbi.nlm.nih.gov/28797616","citation_count":24,"is_preprint":false},{"pmid":"33356381","id":"PMC_33356381","title":"Role of the RNF213 Variant in Vascular Outcomes in Patients With Intracranial Atherosclerosis.","date":"2020","source":"Journal of the American Heart Association","url":"https://pubmed.ncbi.nlm.nih.gov/33356381","citation_count":23,"is_preprint":false},{"pmid":"25817623","id":"PMC_25817623","title":"Genetic Analysis of RNF213 c.14576G>A Variant in Nonatherosclerotic Quasi-Moyamoya Disease.","date":"2015","source":"Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association","url":"https://pubmed.ncbi.nlm.nih.gov/25817623","citation_count":23,"is_preprint":false},{"pmid":"35754359","id":"PMC_35754359","title":"RNF213 loss of function reshapes vascular transcriptome and spliceosome leading to disrupted angiogenesis and aggravated vascular inflammatory responses.","date":"2022","source":"Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/35754359","citation_count":22,"is_preprint":false},{"pmid":"21080342","id":"PMC_21080342","title":"C-MYC rearrangement may induce an aggressive phenotype in anaplastic lymphoma kinase positive anaplastic large cell lymphoma: Identification of a novel fusion gene ALO17/C-MYC.","date":"2011","source":"American journal of hematology","url":"https://pubmed.ncbi.nlm.nih.gov/21080342","citation_count":22,"is_preprint":false},{"pmid":"36611871","id":"PMC_36611871","title":"RNF213 Loss-of-Function Promotes Angiogenesis of Cerebral Microvascular Endothelial Cells in a Cellular State Dependent Manner.","date":"2022","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/36611871","citation_count":22,"is_preprint":false},{"pmid":"28506590","id":"PMC_28506590","title":"RNF213 p.R4810K Variant and Intracranial Arterial Stenosis or Occlusion in Relatives of Patients with Moyamoya Disease.","date":"2017","source":"Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association","url":"https://pubmed.ncbi.nlm.nih.gov/28506590","citation_count":22,"is_preprint":false},{"pmid":"34624841","id":"PMC_34624841","title":"Absence of the RNF213 p.R4810K variant may indicate a severe form of pediatric moyamoya disease in Japanese patients.","date":"2021","source":"Journal of neurosurgery. Pediatrics","url":"https://pubmed.ncbi.nlm.nih.gov/34624841","citation_count":22,"is_preprint":false},{"pmid":"35135845","id":"PMC_35135845","title":"MMD-associated RNF213 SNPs encode dominant-negative alleles that globally impair ubiquitylation.","date":"2022","source":"Life science alliance","url":"https://pubmed.ncbi.nlm.nih.gov/35135845","citation_count":21,"is_preprint":false},{"pmid":"33200540","id":"PMC_33200540","title":"RNF213 gene mutation in circulating tumor DNA detected by targeted next-generation sequencing in the assisted discrimination of early-stage lung cancer from pulmonary nodules.","date":"2020","source":"Thoracic cancer","url":"https://pubmed.ncbi.nlm.nih.gov/33200540","citation_count":21,"is_preprint":false},{"pmid":"38888930","id":"PMC_38888930","title":"RNF213 Variants, Vasospastic Angina, and Risk of Fatal Myocardial Infarction.","date":"2024","source":"JAMA cardiology","url":"https://pubmed.ncbi.nlm.nih.gov/38888930","citation_count":20,"is_preprint":false},{"pmid":"36917666","id":"PMC_36917666","title":"RNF213 modulates γ-herpesvirus infection and reactivation via targeting the viral Replication and Transcription Activator.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/36917666","citation_count":20,"is_preprint":false},{"pmid":"30671466","id":"PMC_30671466","title":"RNF213 Variant Diversity Predisposes Distinct Populations to Dissimilar Cerebrovascular Diseases.","date":"2018","source":"BioMed research international","url":"https://pubmed.ncbi.nlm.nih.gov/30671466","citation_count":20,"is_preprint":false},{"pmid":"28063898","id":"PMC_28063898","title":"The Association of the RNF213 p.R4810K Polymorphism with Quasi-Moyamoya Disease and a Review of the Pertinent Literature.","date":"2017","source":"World neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/28063898","citation_count":20,"is_preprint":false},{"pmid":"33096527","id":"PMC_33096527","title":"Prolonged/delayed cerebral hyperperfusion in adult patients with moyamoya disease with RNF213 gene polymorphism c.14576G>A (rs112735431) after superficial temporal artery-middle cerebral artery anastomosis.","date":"2020","source":"Journal of neurosurgery","url":"https://pubmed.ncbi.nlm.nih.gov/33096527","citation_count":20,"is_preprint":false},{"pmid":"26530008","id":"PMC_26530008","title":"Neuromuscular regulation in zebrafish by a large AAA+ ATPase/ubiquitin ligase, mysterin/RNF213.","date":"2015","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/26530008","citation_count":20,"is_preprint":false},{"pmid":"38243713","id":"PMC_38243713","title":"RNF213 in moyamoya disease: Genotype-phenotype association and the underlying mechanism.","date":"2024","source":"Chinese medical journal","url":"https://pubmed.ncbi.nlm.nih.gov/38243713","citation_count":19,"is_preprint":false},{"pmid":"35543128","id":"PMC_35543128","title":"Association of RNF213 Variants With Periventricular Anastomosis in Moyamoya Disease.","date":"2022","source":"Stroke","url":"https://pubmed.ncbi.nlm.nih.gov/35543128","citation_count":19,"is_preprint":false},{"pmid":"31953610","id":"PMC_31953610","title":"RNF213 suppresses carcinogenesis in glioblastoma by affecting MAPK/JNK signaling pathway.","date":"2020","source":"Clinical & translational oncology : official publication of the Federation of Spanish Oncology Societies and of the National Cancer Institute of Mexico","url":"https://pubmed.ncbi.nlm.nih.gov/31953610","citation_count":19,"is_preprint":false},{"pmid":"31815282","id":"PMC_31815282","title":"Association of single nucleotide polymorphisms of MTHFR, TCN2, RNF213 with susceptibility to hypertension and blood pressure.","date":"2019","source":"Bioscience reports","url":"https://pubmed.ncbi.nlm.nih.gov/31815282","citation_count":19,"is_preprint":false},{"pmid":"26590131","id":"PMC_26590131","title":"Frequency of the moyamoya-related RNF213 p.Arg4810Lys variant in 1,516 Korean individuals.","date":"2015","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/26590131","citation_count":19,"is_preprint":false},{"pmid":"33842849","id":"PMC_33842849","title":"UBC13 is an RNF213-associated E2 ubiquitin-conjugating enzyme, and Lysine 63-linked ubiquitination by the RNF213-UBC13 axis is responsible for angiogenic activity.","date":"2021","source":"FASEB bioAdvances","url":"https://pubmed.ncbi.nlm.nih.gov/33842849","citation_count":18,"is_preprint":false},{"pmid":"29160859","id":"PMC_29160859","title":"The Role of RNF213 4810G>A and 4950G>A Variants in Patients with Moyamoya Disease in Korea.","date":"2017","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/29160859","citation_count":18,"is_preprint":false},{"pmid":"32342250","id":"PMC_32342250","title":"Loss of mitochondrial ClpP, Lonp1, and Tfam triggers transcriptional induction of Rnf213, a susceptibility factor for moyamoya disease.","date":"2020","source":"Neurogenetics","url":"https://pubmed.ncbi.nlm.nih.gov/32342250","citation_count":17,"is_preprint":false},{"pmid":"30925911","id":"PMC_30925911","title":"Rare RNF213 variants and the risk of intracranial artery stenosis/occlusion disease in Chinese population: a case-control study.","date":"2019","source":"BMC medical genetics","url":"https://pubmed.ncbi.nlm.nih.gov/30925911","citation_count":16,"is_preprint":false},{"pmid":"23151810","id":"PMC_23151810","title":"Identification of a novel gene fusion RNF213‑SLC26A11 in chronic myeloid leukemia by RNA-Seq.","date":"2012","source":"Molecular medicine reports","url":"https://pubmed.ncbi.nlm.nih.gov/23151810","citation_count":16,"is_preprint":false},{"pmid":"32248732","id":"PMC_32248732","title":"Mutations of RNF213 are responsible for sporadic cerebral cavernous malformation and lead to a mulberry-like cluster in zebrafish.","date":"2020","source":"Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/32248732","citation_count":16,"is_preprint":false},{"pmid":"26556774","id":"PMC_26556774","title":"Transient middle cerebral artery occlusion in mice induces neuronal expression of RNF213, a susceptibility gene for moyamoya disease.","date":"2015","source":"Brain research","url":"https://pubmed.ncbi.nlm.nih.gov/26556774","citation_count":15,"is_preprint":false},{"pmid":"28617845","id":"PMC_28617845","title":"Frequency and significance of rare RNF213 variants in patients with adult moyamoya disease.","date":"2017","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/28617845","citation_count":15,"is_preprint":false},{"pmid":"26847828","id":"PMC_26847828","title":"The association between the ring finger protein 213 (RNF213) polymorphisms and moyamoya disease susceptibility: a meta-analysis based on case-control studies.","date":"2016","source":"Molecular genetics and genomics : MGG","url":"https://pubmed.ncbi.nlm.nih.gov/26847828","citation_count":14,"is_preprint":false},{"pmid":"31658621","id":"PMC_31658621","title":"The Impact of Moyamoya Disease and RNF213 Mutations on the Spectrum of Plasma Protein and MicroRNA.","date":"2019","source":"Journal of clinical medicine","url":"https://pubmed.ncbi.nlm.nih.gov/31658621","citation_count":14,"is_preprint":false},{"pmid":"36063804","id":"PMC_36063804","title":"Impact of RNF213 c.14576G>A Variant on the Development of Direct and Indirect Revascularization in Pediatric Moyamoya Disease.","date":"2022","source":"Cerebrovascular diseases (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/36063804","citation_count":14,"is_preprint":false},{"pmid":"34955493","id":"PMC_34955493","title":"RNF213 p.R4810K Variant Carriers with Intracranial Arterial Stenosis Have a Low Atherosclerotic Burden.","date":"2021","source":"Journal of atherosclerosis and thrombosis","url":"https://pubmed.ncbi.nlm.nih.gov/34955493","citation_count":14,"is_preprint":false},{"pmid":"31908915","id":"PMC_31908915","title":"A histopathological report of a 16-year-old male with peripheral pulmonary artery stenosis and Moyamoya disease with a homozygous RNF213 mutation.","date":"2019","source":"Respiratory medicine case reports","url":"https://pubmed.ncbi.nlm.nih.gov/31908915","citation_count":14,"is_preprint":false},{"pmid":"37924258","id":"PMC_37924258","title":"De novo variants in RNF213 are associated with a clinical spectrum ranging from Leigh syndrome to early-onset stroke.","date":"2023","source":"Genetics in medicine : official journal of the American College of Medical Genetics","url":"https://pubmed.ncbi.nlm.nih.gov/37924258","citation_count":13,"is_preprint":false},{"pmid":"31064275","id":"PMC_31064275","title":"Genetic analysis of ring finger protein 213 (RNF213) c.14576G>A polymorphism in patients with vertebral artery dissection: a comparative study with moyamoya disease.","date":"2019","source":"Neurological research","url":"https://pubmed.ncbi.nlm.nih.gov/31064275","citation_count":13,"is_preprint":false},{"pmid":"37655297","id":"PMC_37655297","title":"The emerging role of E3 ubiquitin ligase RNF213 as an antimicrobial host determinant.","date":"2023","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/37655297","citation_count":12,"is_preprint":false},{"pmid":"38573771","id":"PMC_38573771","title":"RNF213 variant and autophagic impairment: A pivotal link to endothelial dysfunction in moyamoya disease.","date":"2024","source":"Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/38573771","citation_count":12,"is_preprint":false},{"pmid":"27515544","id":"PMC_27515544","title":"Association between the rs112735431 polymorphism of the RNF213 gene and moyamoya disease: A case-control study and meta-analysis.","date":"2016","source":"Journal of clinical neuroscience : official journal of the Neurosurgical Society of Australasia","url":"https://pubmed.ncbi.nlm.nih.gov/27515544","citation_count":12,"is_preprint":false},{"pmid":"37438553","id":"PMC_37438553","title":"Rnf-213 Knockout Induces Pericyte Reduction and Blood-Brain Barrier Impairment in Mouse.","date":"2023","source":"Molecular neurobiology","url":"https://pubmed.ncbi.nlm.nih.gov/37438553","citation_count":12,"is_preprint":false},{"pmid":"34804992","id":"PMC_34804992","title":"Proteome Profiling of RNF213 Depleted Cells Reveals Nitric Oxide Regulator DDAH1 Antilisterial Activity.","date":"2021","source":"Frontiers in cellular and infection microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/34804992","citation_count":12,"is_preprint":false},{"pmid":"34373710","id":"PMC_34373710","title":"RNF213 gene silencing upregulates transforming growth factor β1 expression in bone marrow-derived mesenchymal stem cells and is involved in the onset of Moyamoya disease.","date":"2021","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34373710","citation_count":12,"is_preprint":false},{"pmid":"35876407","id":"PMC_35876407","title":"RNF213 R4810K Variant in Suspected Unilateral Moyamoya Disease Predicts Contralateral Progression.","date":"2022","source":"Journal of the American Heart Association","url":"https://pubmed.ncbi.nlm.nih.gov/35876407","citation_count":12,"is_preprint":false},{"pmid":"28734662","id":"PMC_28734662","title":"Transient Global Cerebral Ischemia Induces RNF213, a Moyamoya Disease Susceptibility Gene, in Vulnerable Neurons of the Rat Hippocampus CA1 Subregion and Ischemic Cortex.","date":"2017","source":"Journal of stroke and cerebrovascular diseases : the official journal of National Stroke Association","url":"https://pubmed.ncbi.nlm.nih.gov/28734662","citation_count":12,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":57352,"output_tokens":9198,"usd":0.155013,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":19364,"output_tokens":4909,"usd":0.109772,"stage2_stop_reason":"end_turn"},"total_usd":0.264785,"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\": 2011,\n      \"finding\": \"RNF213 encodes a 591-kDa cytosolic protein possessing a Walker motif (ATPase) and a RING finger domain that exhibit ATPase and ubiquitin ligase activities, respectively. Knockdown of RNF213 in zebrafish caused irregular wall formation in trunk arteries and abnormal sprouting vessels, establishing a role in vascular development.\",\n      \"method\": \"Exome sequencing/Sanger sequencing for variant identification; functional assays for ATPase/ubiquitin ligase activity; zebrafish morpholino knockdown with vascular phenotype readout\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — biochemical activity assays combined with in vivo zebrafish loss-of-function with defined vascular phenotype; foundational paper replicated across many subsequent studies\",\n      \"pmids\": [\"21799892\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Mysterin/RNF213 contains two tandem AAA+ ATPase modules and forms a large ring-shaped oligomeric complex. Fluorescence correlation spectroscopy and biochemical evaluation showed that RNF213 dynamically changes its oligomeric state through ATP/ADP binding and hydrolysis cycles, consistent with a mechanical AAA+ ATPase mechanism.\",\n      \"method\": \"Fluorescence correlation spectroscopy (FCS), biochemical sedimentation/native PAGE assays for oligomeric state, ATPase activity assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical reconstitution with multiple orthogonal methods (FCS + biochemical assays) in a single study\",\n      \"pmids\": [\"24658080\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"RNF213 is upregulated by IFN-β through STAT signaling in endothelial cells and mediates antiangiogenic activity of IFN-β. The R4810K variant decreases ATPase activity (similar to a Walker B motif stabilizing mutation WEQ) and stabilizes oligomers, causing antiangiogenic activity even without IFN-β stimulation. EC-specific Rnf213 R4757K transgenic mice showed suppressed cerebral angiogenesis under hypoxia, while wild-type overexpression did not.\",\n      \"method\": \"Promoter analysis (STAT binding), ATPase activity assays, mutagenesis of Walker B motif, transgenic mouse hypoxia model with in vivo angiogenesis readout\",\n      \"journal\": \"Journal of the American Heart Association\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro biochemical assays with mutagenesis plus in vivo transgenic mouse model with functional angiogenesis readout\",\n      \"pmids\": [\"26126547\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Pro-inflammatory cytokines IFN-γ and TNF-α synergistically activate transcription of RNF213 in endothelial cells via AKT and PKR pathways. RNF213 knockdown in endothelial cells reduced expression of cell cycle-promoting genes, decreased proliferation, reduced angiogenic capacity, and down-regulated matrix metalloproteases specifically in endothelial cells.\",\n      \"method\": \"Chemical inhibitor experiments (LY294002 for AKT, C16 for PKR), RNAi knockdown, transcriptome-wide analysis, qPCR validation, proliferation and tube formation assays\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (transcriptomics + functional assays + inhibitor studies) in a single lab\",\n      \"pmids\": [\"26278786\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"RNF213 R4810K variant downregulates Securin in iPSC-derived vascular endothelial cells, inhibiting angiogenic activity. Overexpression of R4810K (but not wild-type) RNF213 reduced tube formation and proliferation in HUVECs. siRNA knockdown of Securin phenocopied the angiogenic defect.\",\n      \"method\": \"iPSC differentiation to endothelial cells, gene expression profiling, overexpression of WT vs. R4810K in HUVECs, tube formation assay, siRNA knockdown of Securin\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (iPSC model + overexpression + siRNA) establishing substrate/pathway link\",\n      \"pmids\": [\"23850618\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"RNF213 R4810K variant induces mitotic abnormalities: overexpression in HeLa cells extended mitosis 4-fold, and co-immunoprecipitation revealed that R4810K forms a complex with MAD2 more readily than wild-type RNF213. Fibroblasts and iPSC-derived endothelial cells from patients showed desynchronized MAD2 localization and increased aneuploidy.\",\n      \"method\": \"Live-cell imaging, co-immunoprecipitation, immunofluorescence, flow cytometry for aneuploidy, MAD2 depletion epistasis experiment\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP identifying RNF213/MAD2 complex plus functional mitotic phenotype with epistasis (MAD2 depletion)\",\n      \"pmids\": [\"23994138\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PTP1B negatively regulates RNF213 in HER2+ breast cancer cells. RNF213 knockdown reverses the effects of PTP1B deficiency on α-KG-dependent dioxygenase (α-KGDD) activity and non-mitochondrial oxygen consumption, establishing a PTP1B→RNF213→α-KGDD pathway required for tumor survival under hypoxia.\",\n      \"method\": \"RNF213 siRNA knockdown in PTP1B-deficient and wild-type HER2+ breast cancer cells, oxygen consumption measurements, α-KGDD activity assays, xenograft tumorigenicity rescue\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — epistasis established by knockdown rescue, multiple orthogonal functional readouts (NMOC, α-KGDD, xenograft growth), replicated across multiple cell lines\",\n      \"pmids\": [\"27323329\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Cryo-EM structure of mouse RNF213 (584 kDa) revealed an N-terminal stalk, a dynein-like core with six ATPase units, and a multidomain E3 module. Collaboration with UbcH7, a cysteine-reactive E2, indicates a RING-independent ubiquitin-transfer mechanism. Pathogenic MMD mutations cluster in the composite E3 domain.\",\n      \"method\": \"Cryo-EM structure determination, biochemical E2 collaboration assays with UbcH7, mutational mapping of disease variants onto structure\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — cryo-EM structure with biochemical validation of ubiquitin transfer mechanism; landmark structural paper\",\n      \"pmids\": [\"32573437\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RING domain mutations from MMD patients reduce RNF213 ubiquitin ligase activity (using Ubc13/Uev1A as E2, generating K63-linked polyubiquitin chains in vitro). In full-length overexpression experiments in HEK293T cells, these mutations conversely enhanced NF-κB activation and caspase-3-mediated apoptosis in an AAA+ domain-dependent manner, suggesting NF-κB/apoptosis activities are negatively regulated by RNF213 E3 ligase activity.\",\n      \"method\": \"In vitro ubiquitination assays with purified RING domain, K63 linkage-specific antibodies, full-length overexpression in HEK293T, NF-κB reporter assay, caspase-3 activity assay, AAA+ domain deletion mutants\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro ubiquitination assay with linkage specificity plus cell-based functional studies; single lab\",\n      \"pmids\": [\"32139119\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF213 ubiquitylates the lipid A moiety of bacterial lipopolysaccharide (LPS) on cytosol-invading Salmonella, forming a ubiquitin coat. This requires the dynein-like core of RNF213 but not its RING domain; instead, an RZ finger mediates ubiquitylation. RNF213 is essential for recruitment of LUBAC (which adds M1-linked chains) and ubiquitin-dependent autophagy receptors, restricting Salmonella proliferation.\",\n      \"method\": \"Biochemical ubiquitylation assays identifying LPS as substrate, domain deletion/mutagenesis (RZ finger vs. RING domain), co-immunoprecipitation for LUBAC recruitment, bacterial colony-forming unit assays, autophagy receptor recruitment imaging\",\n      \"journal\": \"Nature\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — reconstitution of LPS ubiquitylation with mechanistic domain mutagenesis, multiple functional readouts; landmark mechanistic paper\",\n      \"pmids\": [\"34012115\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"UBC13 (UBE2N) is an E2 ubiquitin-conjugating enzyme for RNF213, identified by yeast two-hybrid screening with the RNF213 RING domain as bait, and confirmed by co-immunoprecipitation in vivo. RNF213 autoubiquitinates via K63-linked chains (not K48) in a UBC13-dependent manner. UBC13 knockdown and ubiquitination-dead RNF213 mutants reduced HUVEC cell migration and invasion, indicating the RNF213-UBC13 axis contributes to angiogenic activity.\",\n      \"method\": \"Yeast two-hybrid screening, co-immunoprecipitation in vivo, in vitro ubiquitination assay with K63/K48 linkage analysis, UBC13 siRNA knockdown, migration/invasion assays\",\n      \"journal\": \"FASEB bioAdvances\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — yeast two-hybrid plus Co-IP for binding, in vitro ubiquitination for mechanism, functional rescue experiments; single lab\",\n      \"pmids\": [\"33842849\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF213 uses E2-conjugating enzymes UBE2D2 and UBE2L3 for distinct ubiquitylation events: RNF213-UBE2D2 catalyzes K6 (and K48) poly-ubiquitylation in vitro, while RNF213-UBE2L3 catalyzes K6-, K11-, and K48-linked chains. MMD-associated SNPs encode proteins with decreased E3 activity. The most frequent MMD allele (p.R4810K) acts as a dominant-negative mutant that globally decreases ubiquitylation but does not affect ATPase activity.\",\n      \"method\": \"In vitro ubiquitination assays with purified components, ubiquitin linkage analysis, dominant-negative functional assay in cells, ATPase activity assays for MMD SNP alleles\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — rigorous in vitro reconstitution with multiple E2 enzymes, linkage specificity determined, mechanistic characterization of disease variants; multiple orthogonal methods\",\n      \"pmids\": [\"35135845\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF213 knockout in human cerebral endothelial cells (CRISPR-Cas9) caused morphological changes, increased blood-brain barrier permeability, downregulation and delocalization of PECAM-1, abnormal leukocyte transmigration, and secretion of proinflammatory cytokines, identifying RNF213 as a regulator of cerebral endothelium integrity.\",\n      \"method\": \"CRISPR-Cas9 knockout in hCMEC/D3 cells, permeability assays, immunofluorescence for junction proteins (PECAM-1), transendothelial leukocyte migration assay, cytokine ELISA\",\n      \"journal\": \"Stroke\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean CRISPR knockout with multiple orthogonal functional readouts; single lab\",\n      \"pmids\": [\"34991336\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"IFN-γ-inducible RNF213 translocates to Toxoplasma gondii parasitophorous vacuoles (PVs) and facilitates PV ubiquitylation (linear and K63-linked) in human cells independently of LUBAC. RNF213 is required for IFN-γ-mediated growth restriction of T. gondii. The ATPase domain is required for RNF213 recruitment to the PVM, while loss of a critical histidine in the RZ finger abolishes ubiquitylation; both are needed for parasite growth restriction.\",\n      \"method\": \"CRISPR loss-of-function screen, domain mutagenesis (ATPase domain, RZ finger histidine), immunofluorescence for PV localization, ubiquitin linkage analysis, intracellular parasite growth assay\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — domain mutagenesis establishing distinct roles of ATPase (recruitment) vs. RZ finger (ubiquitylation), functional growth restriction assay, confirmed independently across studies\",\n      \"pmids\": [\"36154443\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"GarD, a Chlamydia trachomatis inclusion membrane protein, functions as a cis-acting stealth factor that bars IFN-γ-inducible RNF213 from targeting inclusions. In IFN-γ-primed human cells lacking garD, inclusions become decorated with linear ubiquitin in an RNF213-dependent manner and are destroyed, establishing RNF213 as an anti-Chlamydia E3 ligase.\",\n      \"method\": \"Genetic screen, garD loss-of-function mutants, RNF213 knockdown/knockout, immunofluorescence for ubiquitin on inclusions, bacterial growth assays\",\n      \"journal\": \"Cell host & microbe\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genetic screen plus functional validation with bacterial mutants and host RNF213 knockdown; mechanistic antagonism defined\",\n      \"pmids\": [\"36084633\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF213 directly interacts with the Replication and Transcription Activator (RTA) of KSHV and MHV-68, promotes K48-linked ubiquitination of RTA, and targets it for proteasome-dependent degradation, thereby inhibiting gammaherpesvirus de novo infection and lytic reactivation.\",\n      \"method\": \"Co-immunoprecipitation (RNF213-RTA interaction), in vitro ubiquitination assay (K48 linkage), proteasome inhibitor experiments, viral infection/reactivation assays with RNF213 overexpression and knockdown\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — direct binding (Co-IP) plus in vitro ubiquitination with linkage specificity plus proteasomal degradation mechanism plus functional viral restriction; multiple orthogonal methods in single study\",\n      \"pmids\": [\"36917666\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Genome-wide CRISPR screen identified RNF213 as necessary for IFN-γ-mediated restriction of T. gondii. The ATPase domain is required for RNF213 recruitment to the parasitophorous vacuole membrane (PVM), while the RZ finger domain is required for ubiquitination. Both are required for parasite growth restriction. RNF213-mediated restriction is independent of ATG5 (non-canonical autophagy). RNF213 overexpression in naive (non-IFN-γ stimulated) cells is sufficient to restrict T. gondii growth.\",\n      \"method\": \"Genome-wide CRISPR screen, targeted ISG CRISPR screen, ATPase and RZ finger domain mutational analysis, fluorescence imaging of PVM localization, intracellular growth assays, ATG5 knockout epistasis\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — genome-wide screen plus domain mutagenesis plus epistasis with ATG5; multiple orthogonal methods establishing mechanism\",\n      \"pmids\": [\"38147552\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF213 loss-of-function activates the Hippo pathway effector YAP/TAZ, promoting overexpression of VEGFR2 downstream. Inhibition of YAP/TAZ altered VEGFR2 trafficking from Golgi to plasma membrane and reversed RNF213 knockdown-induced pathological angiogenesis in endothelial cells and in vivo (RNF213-deficient mice and zebrafish).\",\n      \"method\": \"RNF213 knockdown in human brain microvascular endothelial cells, bulk RNA-seq and single-cell RNA-seq, YAP/TAZ inhibitor experiments, VEGFR2 trafficking assay (Golgi vs. plasma membrane fractionation), in vivo RNF213-KO mouse and zebrafish vascular phenotyping\",\n      \"journal\": \"Brain : a journal of neurology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pathway inhibitor epistasis plus transcriptomics plus in vivo validation; single lab\",\n      \"pmids\": [\"37399508\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RNF213 specifically promotes regulatory T (Treg) cell differentiation and attenuates autoimmune disease in an FOXO1-dependent manner. Mechanistically, RNF213 interacts with FOXO1 and promotes its nuclear translocation via K63-linked ubiquitination. RNF213 expression in CD4+ T cells is induced by IFN-β and is required for IFN-β therapeutic efficacy in multiple sclerosis models.\",\n      \"method\": \"Co-immunoprecipitation (RNF213-FOXO1 interaction), K63 ubiquitination assays, nuclear fractionation, Treg differentiation assays, autoimmune disease mouse models, IFN-β treatment experiments\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP establishing binding, ubiquitination assay with linkage specificity, nuclear translocation assay, functional Treg differentiation rescue; multiple orthogonal methods\",\n      \"pmids\": [\"39013878\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"S-nitrosylation of RNF213 was detected in the P301S tauopathy mouse model cortex and hippocampus. S-nitrosylated RNF213 was associated with increased levels of NFAT-1 and FILAMIN-A, implicating RNF213 S-nitrosylation in activation of non-canonical Wnt/Ca²⁺ signaling in tauopathy.\",\n      \"method\": \"S-nitrosoproteome mass spectrometry, pathway analysis, immunoblotting for downstream effectors (pCaMKII, NFAT-1, FILAMIN-A) in transgenic mouse brain tissue\",\n      \"journal\": \"Translational psychiatry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — proteomics identification of SNO-RNF213 with correlative downstream measurements; no direct functional validation of the modification on RNF213 activity\",\n      \"pmids\": [\"30696811\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Genetic ablation of mitochondrial matrix factors ClpP, Tfam, and Lonp1 potently induces Rnf213 transcript expression in multiple organs. Rnf213 is also induced by Poly(I:C)-triggered TLR3-mediated responses and IFN-γ treatment. This induction is only partially suppressed by PKR antagonist C16, suggesting RNF213 is induced by mitochondrial dysfunction/dsRNA-dependent inflammation.\",\n      \"method\": \"Rnf213 mRNA quantification in tissue from genetic KO mice (ClpP, Tfam, Lonp1), Poly(I:C) treatment, IFN-γ treatment, PKR inhibitor (C16) experiments\",\n      \"journal\": \"Neurogenetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple genetic KO models plus pharmacological inhibitor experiments establishing upstream induction pathway; single lab\",\n      \"pmids\": [\"32342250\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RNF213-deficient mice show suppressed vascular remodeling after common carotid artery ligation: the intima and medial layers were significantly thinner in RNF213-/- than wild-type mice after ligation. Under normal conditions, no spontaneous arterial abnormalities were observed up to 64 weeks, indicating RNF213 loss is insufficient alone to cause moyamoya disease.\",\n      \"method\": \"Cre-lox RNF213 knockout mouse generation, 9.4-T MRA, common carotid artery ligation model, Elastica-Masson staining of arterial wall\",\n      \"journal\": \"Brain research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse with defined vascular phenotype using quantitative histopathology and MRA; single lab\",\n      \"pmids\": [\"24440776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RNF213-deficient mice showed enhanced angiogenesis and improved blood flow recovery after permanent femoral artery ligation compared to wild-type, indicating RNF213 functions to suppress angiogenesis under chronic ischemic conditions.\",\n      \"method\": \"RNF213-/- knockout mice, femoral artery ligation model, laser speckle flowmetry, vascular density quantification, ambulatory impairment scoring\",\n      \"journal\": \"Brain research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse with multiple functional readouts (blood flow, vascular density, behavioral); single lab\",\n      \"pmids\": [\"25446450\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"RNF213-deficient mice showed significantly increased vascular MMP-9 expression at 1 and 7 days after common carotid artery ligation, and thinner vascular walls at 14 days, linking RNF213 loss to dysregulated MMP-9-dependent vascular remodeling.\",\n      \"method\": \"RNF213-/- knockout mice, common carotid artery ligation, immunohistochemistry/immunoblotting for MMP-9, Elastica-Masson staining\",\n      \"journal\": \"Neuroreport\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse with defined molecular and histological phenotype; single lab\",\n      \"pmids\": [\"25383461\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"Mysterin/RNF213 is required for proper fast muscle formation and neuromuscular regulation in zebrafish. Morpholino-mediated knockdown caused significant reduction in fast myofibrils and immature projection of primary motoneurons, leading to severe motor deficits. Rescue with fast muscle-specific RNF213 expression reversed these phenotypes. Both AAA+ ATPase and ubiquitin ligase activities were indispensable for proper fast muscle formation.\",\n      \"method\": \"Zebrafish antisense morpholino knockdown, tissue-specific mRNA rescue, immunofluorescence for myofibrils and motoneuron projections, behavioral motor assays, ATPase-dead and RING-dead rescue constructs\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — loss-of-function with tissue-specific rescue, activity-dead mutagenesis establishing both enzymatic domains are necessary; multiple orthogonal readouts\",\n      \"pmids\": [\"26530008\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"RNF213 is required for maintenance of cerebral blood flow under hypoperfusion. RNF213 knockout mice showed significantly more severe CBF reduction and impaired CBF restoration compared to wild-type after bilateral carotid stenosis. EC-specific Rnf213 mutant (R4810K ortholog) transgenic mice also showed impaired CBF restoration and reduced angiogenesis, establishing a vascular endothelial cell-autonomous role.\",\n      \"method\": \"Bilateral carotid artery stenosis surgery, arterial spin-labeling MRI for quantitative CBF measurements, histological angiogenesis analysis in RNF213 KO and EC-specific transgenic mice\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple mouse models (KO and EC-specific transgenic) with quantitative CBF and histological readouts; single lab\",\n      \"pmids\": [\"29483617\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF213 is required for resistance to Rift Valley fever virus in mice. CRISPR/Cas9 Rnf213-deficient C57BL/6 mice were significantly more susceptible to RVF than controls, with reduced average survival time post-infection.\",\n      \"method\": \"CRISPR/Cas9 Rnf213 knockout mouse, Rift Valley fever virus infection, survival analysis\",\n      \"journal\": \"Mammalian genome\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean CRISPR KO mouse with defined in vivo infectious disease phenotype; single lab\",\n      \"pmids\": [\"33420513\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF213 knockdown in macrophages and adipocytes reduces its expression, which is enhanced by pro-inflammatory TNF-α stimuli (LPS) and suppressed by PPARγ-mediated anti-inflammatory stimuli. PTP1B inactivation abolished RNF213 expression and enhanced adipogenesis through PPARγ. Constitutive RNF213 expression suppressed adipocyte differentiation by inhibiting PPARγ, establishing a TNF-α/PTP1B/RNF213/PPARγ pathway in adipogenesis.\",\n      \"method\": \"RNF213 knockdown in macrophages/adipocytes, LPS stimulation, PPARγ inhibitor/activator treatment, PTP1B inhibitor experiments, adipogenesis assays (Oil Red O staining), qPCR\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — predominantly transcriptional/expression-level readouts without direct biochemical demonstration of pathway hierarchy; single lab\",\n      \"pmids\": [\"33333224\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF213 knockdown in bone marrow-derived mesenchymal stem cells upregulates TGF-β1 at both protein and mRNA levels but does not affect VEGF expression, suggesting RNF213 normally suppresses TGF-β1 in these cells.\",\n      \"method\": \"Lentivirus-mediated shRNA knockdown of RNF213 in rat BMSCs, RT-qPCR, ELISA for TGF-β1 and VEGF expression\",\n      \"journal\": \"Experimental and therapeutic medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single knockdown experiment with transcriptional/protein readout only; no mechanistic pathway established; single lab\",\n      \"pmids\": [\"34373710\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"RNF213 knockdown in HUVECs disrupts angiogenesis partly through downregulation of DNA replication/proliferation pathways. Endothelial cells sensitized to LPS-induced inflammation after RNF213 KD showed retarded migration and enhanced macrophage transmigration. RNF213 KD also caused extensive changes in mRNA splicing. In vascular smooth muscle cells, RNF213 KD altered cytoskeletal organization and contractility.\",\n      \"method\": \"RNF213 siRNA knockdown in HUVECs and vSMCs, transcriptome sequencing, splicing analysis, angiogenesis assays, LPS stimulation, macrophage transmigration assays, co-culture models\",\n      \"journal\": \"Journal of cerebral blood flow and metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — comprehensive transcriptome plus multiple functional assays across two cell types; single lab\",\n      \"pmids\": [\"35754359\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"RNF213 knockout in mice causes significant pericyte reduction, blood-brain barrier impairment in cortex, microglia activation, elevated proinflammatory cytokines, and reduced tight junction proteins (Occludin, Claudin-5, ZO-1).\",\n      \"method\": \"Rnf213 knockout mice, immunofluorescence for pericyte markers and tight junction proteins, cytokine quantification, BBB permeability assays\",\n      \"journal\": \"Molecular neurobiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO mouse with multiple cellular phenotype readouts; single lab\",\n      \"pmids\": [\"37438553\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"RNF213 R4810K variant-expressing HUVECs show autophagy inhibition (increased SQSTM1/p62 and LC3-II) and impaired endothelial function (tube formation) after oxygen-glucose deprivation (OGD). Rapamycin and cilostazol as autophagy inducers restored RNF213 R4810K cell function, linking R4810K to autophagic impairment under ischemic stress.\",\n      \"method\": \"Transfection of RNF213 WT vs. R4810K in HUVECs, OGD model, immunoblotting for autophagy markers (p62, LC3-II), tube formation assay, transmission electron microscopy for autophagic vesicles, rapamycin/cilostazol pharmacological rescue\",\n      \"journal\": \"Journal of cerebral blood flow and metabolism\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — WT vs. mutant comparison with pharmacological rescue and multiple readouts including EM; single lab\",\n      \"pmids\": [\"38573771\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"RNF213 controls Listeria monocytogenes infection through regulation of DDAH1 transcription and production of nitric oxide (NO). RNF213 knockdown downregulates DDAH1, reducing NO production in macrophages from RNF213 KO mice, thereby impairing anti-Listeria defense.\",\n      \"method\": \"Mass spectrometry-based proteomics of RNF213-depleted cells, RT-qPCR validation, DDAH1 knockdown, NO production measurements, Listeria growth assays in RNF213 KO mouse macrophages\",\n      \"journal\": \"Frontiers in cellular and infection microbiology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — proteomics plus functional validation with KO mouse macrophages and pathway mechanism (DDAH1-NO axis); single lab\",\n      \"pmids\": [\"34804992\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"RNF213 is a 591 kDa cytosolic E3 ubiquitin ligase and AAA+ ATPase with a dynein-like six-unit ATPase core and a multidomain E3 module (including an RZ finger critical for ubiquitylation and a RING domain); it ubiquitylates non-proteinaceous substrates such as bacterial LPS via its RZ finger (RING-independently), ubiquitylates protein substrates (e.g., KSHV/MHV-68 RTA via K48 linkage for proteasomal degradation, FOXO1 via K63 linkage for nuclear translocation) through its RING domain in collaboration with multiple E2 enzymes (UBC13/UBE2N, UBE2D2, UBE2L3, UbcH7), is transcriptionally induced by IFN-γ, IFN-β, TNF-α, and cellular stress (mitochondrial dysfunction/TLR3 activation), and acts as an innate immune effector that restricts cytosolic bacteria (Salmonella, Listeria), parasites (Toxoplasma), and viruses (KSHV, MHV-68) by ubiquitin-coating pathogen-containing compartments; in vascular endothelial cells it regulates angiogenesis, blood-brain barrier integrity, and cerebral blood flow, and pathogenic MMD-associated variants (especially p.R4810K) act as dominant-negative alleles that globally impair ubiquitylation and destabilize RNF213 ATPase oligomerization, thereby dysregulating angiogenic and inflammatory signaling.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"RNF213 is a giant (~591 kDa) cytosolic protein that couples a dynein-like AAA+ ATPase core to a multidomain E3 ubiquitin ligase module, functioning both as an innate immune effector and as a regulator of vascular homeostasis [#0, #7]. Its ATPase core forms a ring-shaped oligomer whose assembly is dynamically remodeled through ATP/ADP cycles [#1], while its E3 module catalyzes ubiquitylation through two mechanistically distinct routes: a RING-independent RZ-finger mechanism that ubiquitylates the lipid A moiety of bacterial LPS, and a RING-dependent mechanism operating with multiple E2 enzymes (UBC13/UBE2N, UBE2D2, UBE2L3, UbcH7) to build diverse chain linkages [#7, #9, #10, #11]. As an interferon-inducible effector, RNF213 is recruited to pathogen-containing compartments—Salmonella, Toxoplasma parasitophorous vacuoles, and Chlamydia inclusions—where ATPase-dependent recruitment and RZ-finger-dependent ubiquitylation coat the surface in ubiquitin and recruit downstream LUBAC and autophagy machinery to restrict the pathogen [#9, #13, #16, #14]; it likewise targets viral proteins such as gammaherpesvirus RTA for K48-linked proteasomal degradation [#15] and promotes Treg differentiation via K63-linked ubiquitylation and nuclear translocation of FOXO1 [#18]. In vascular endothelium RNF213 regulates angiogenesis, cerebral blood flow, and blood-brain barrier integrity [#22, #25, #30], acting in part through the UBC13 ubiquitylation axis [#10] and through suppression of YAP/TAZ-VEGFR2 signaling [#17]. Transcription of RNF213 is induced by IFN-\\u03b2, IFN-\\u03b3, and TNF-\\u03b1 as well as by mitochondrial dysfunction and TLR3 activation [#2, #3, #20]. The major moyamoya disease-associated allele p.R4810K behaves as a dominant-negative that globally impairs ubiquitylation and disrupts endothelial angiogenic function [#11, #2, #4].\",\n  \"teleology\": [\n    {\n      \"year\": 2011,\n      \"claim\": \"Established RNF213 as a dual-activity enzyme and linked it to vascular development, framing the central question of how a single protein bridges enzymology and angiogenesis.\",\n      \"evidence\": \"Biochemical ATPase/ubiquitin ligase assays plus zebrafish morpholino knockdown with vascular readout\",\n      \"pmids\": [\"21799892\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No substrates identified\", \"Mechanistic link between enzymatic activity and vascular phenotype undefined\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Showed the AAA+ core assembles into a dynamic ring-shaped oligomer driven by ATP/ADP cycling, defining RNF213 as a mechanical AAA+ ATPase.\",\n      \"evidence\": \"Fluorescence correlation spectroscopy and biochemical oligomeric-state and ATPase assays\",\n      \"pmids\": [\"24658080\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of oligomeric remodeling unresolved\", \"No structure available at this stage\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"In vivo mouse models established that RNF213 loss suppresses vascular remodeling and enhances ischemic angiogenesis while being insufficient alone to cause arterial disease, defining its role as a modulator rather than sole driver.\",\n      \"evidence\": \"RNF213 knockout mice in carotid ligation and femoral ligation models with MRA, flowmetry, and MMP-9 analysis\",\n      \"pmids\": [\"24440776\", \"25446450\", \"25383461\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular substrate linking RNF213 to remodeling unidentified\", \"MMP-9 regulation mechanism indirect\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Connected RNF213 to interferon and inflammatory signaling, showing it mediates IFN-\\u03b2 antiangiogenic activity and is co-induced by IFN-\\u03b3/TNF-\\u03b1, establishing it as a cytokine-responsive endothelial effector.\",\n      \"evidence\": \"Promoter/STAT analysis, AKT/PKR inhibitor studies, ATPase assays, transgenic mouse hypoxia angiogenesis model, RNAi with proliferation/tube formation readouts\",\n      \"pmids\": [\"26126547\", \"26278786\", \"23850618\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct ubiquitylation substrate in angiogenesis not defined\", \"Securin link correlative\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Placed RNF213 in a PTP1B-dependent metabolic pathway controlling \\u03b1-KG-dependent dioxygenase activity and hypoxic tumor survival, extending its role beyond vasculature.\",\n      \"evidence\": \"siRNA knockdown rescue in HER2+ breast cancer cells, oxygen consumption and \\u03b1-KGDD assays, xenograft tumorigenicity\",\n      \"pmids\": [\"27323329\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct enzymatic substrate in this pathway unknown\", \"Mechanism of \\u03b1-KGDD regulation indirect\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"The cryo-EM structure revealed the N-terminal stalk, six-unit dynein-like ATPase core, and composite E3 module, and uncovered a RING-independent, cysteine-reactive (UbcH7) ubiquitin-transfer mechanism while mapping disease mutations to the E3 region.\",\n      \"evidence\": \"Cryo-EM structure of mouse RNF213 with E2 collaboration assays and disease-variant mapping\",\n      \"pmids\": [\"32573437\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Substrate engagement geometry not resolved\", \"Coupling between ATPase cycling and ubiquitin transfer not structurally defined\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Defined the E2 repertoire and linkage outputs and demonstrated that disease alleles, particularly p.R4810K, act as dominant-negatives that globally impair ubiquitylation without altering ATPase activity, clarifying how variants cause loss of E3 function.\",\n      \"evidence\": \"In vitro ubiquitination with UBC13/Uev1A, UBE2D2, UBE2L3, linkage analysis, NF-\\u03baB and caspase reporters, dominant-negative cell assays, ATPase assays on disease alleles\",\n      \"pmids\": [\"32139119\", \"35135845\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Physiological substrate(s) for each E2-linkage combination not all identified\", \"How dominant-negative impairment maps to vascular pathology incomplete\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Identified upstream inducers beyond classical cytokines, showing mitochondrial matrix dysfunction and TLR3/dsRNA signaling drive Rnf213 expression, broadening its stress-response regulation.\",\n      \"evidence\": \"Rnf213 mRNA quantification in ClpP/Tfam/Lonp1 KO mice, Poly(I:C), IFN-\\u03b3, and PKR inhibitor experiments\",\n      \"pmids\": [\"32342250\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Transcription factors mediating mitochondrial-stress induction undefined\", \"Functional output of stress-induced RNF213 not established\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Defined RNF213 as a direct innate immune effector that ubiquitylates non-proteinaceous LPS lipid A on cytosolic bacteria via its RZ finger (RING-independent), seeding LUBAC and autophagy recruitment to restrict pathogens.\",\n      \"evidence\": \"Biochemical ubiquitylation of LPS, RZ-finger vs RING domain mutagenesis, LUBAC Co-IP, autophagy receptor imaging, CFU assays on Salmonella; plus UBC13 yeast two-hybrid/Co-IP and autoubiquitylation analysis; plus Listeria DDAH1-NO axis and RVFV resistance models\",\n      \"pmids\": [\"34012115\", \"33842849\", \"34804992\", \"33420513\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How RNF213 recognizes diverse pathogen surfaces unclear\", \"Relationship between LPS ubiquitylation and protein-substrate ubiquitylation not unified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Generalized RNF213 pathogen restriction to eukaryotic parasites and other bacteria, showing ATPase-dependent recruitment and RZ-finger-dependent ubiquitylation of Toxoplasma vacuoles and Chlamydia inclusions, with pathogen stealth factors (GarD) antagonizing it.\",\n      \"evidence\": \"Genome-wide and ISG CRISPR screens, domain mutagenesis (ATPase recruitment vs RZ ubiquitylation), PVM/inclusion imaging, ATG5 epistasis, garD bacterial mutants, growth restriction assays\",\n      \"pmids\": [\"36154443\", \"38147552\", \"36084633\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Signal directing RNF213 to specific compartments unknown\", \"Downstream effector requirements vary across pathogens\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended RNF213's E3 function to protein substrates in immunity, showing K48-linked degradation of gammaherpesvirus RTA to restrict viral lytic reactivation.\",\n      \"evidence\": \"Co-IP, in vitro K48 ubiquitination, proteasome inhibitor experiments, viral infection/reactivation assays\",\n      \"pmids\": [\"36917666\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether RING vs RZ finger drives RTA ubiquitylation not delineated\", \"E2 partner for this event unspecified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Linked endothelial RNF213 loss to pathological angiogenesis via YAP/TAZ-driven VEGFR2 overexpression and trafficking, and to blood-brain barrier integrity through pericyte and tight-junction regulation.\",\n      \"evidence\": \"RNF213 knockdown with RNA-seq/scRNA-seq, YAP/TAZ inhibitor epistasis, VEGFR2 trafficking fractionation, in vivo mouse/zebrafish phenotyping; plus Rnf213 KO mouse BBB/pericyte analysis\",\n      \"pmids\": [\"37399508\", \"37438553\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ubiquitylation substrate upstream of YAP/TAZ unidentified\", \"Connection to immune effector function unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a substrate-level immunoregulatory role, showing RNF213 promotes Treg differentiation by K63-ubiquitylating FOXO1 to drive its nuclear translocation and is required for IFN-\\u03b2 therapeutic efficacy.\",\n      \"evidence\": \"Co-IP, K63 ubiquitination assays, nuclear fractionation, Treg differentiation and autoimmune disease mouse models, IFN-\\u03b2 treatment\",\n      \"pmids\": [\"39013878\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether FOXO1 ubiquitylation uses RING or RZ finger not stated\", \"Structural basis of K63 vs K48 substrate selection unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How RNF213's two enzymatic activities are mechanistically coupled, and how a single ligase integrates pathogen restriction, vascular homeostasis, and immunometabolic signaling through distinct substrates and linkage types, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying model connecting ATPase cycling to substrate-specific ubiquitylation\", \"The full physiological substrate set is incomplete\", \"Causal mechanism by which p.R4810K produces moyamoya pathology in vivo not established\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 7, 9, 10, 11, 15, 18]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [9, 15, 18]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [0, 1, 7, 13]},\n      {\"term_id\": \"GO:0016874\", \"supporting_discovery_ids\": [0, 7, 11]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [9, 13, 14, 15, 16, 18]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [9, 11, 15, 18]},\n      {\"term_id\": \"R-HSA-1266738\", \"supporting_discovery_ids\": [0, 17, 22, 25]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"UBE2N\", \"UBE2D2\", \"UBE2L3\", \"FOXO1\", \"MAD2\", \"PTP1B\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}