{"gene":"GBP3","run_date":"2026-04-28T18:06:52","timeline":{"discoveries":[{"year":1998,"finding":"Mouse GBP3 (mGBP3) is a 71-kDa guanylate-binding protein with intrinsic GTPase activity (Km 77 µM, Vmax 21 pmol/min/µg), binds agarose-immobilized guanine nucleotides (GTP, GDP, GMP), lacks the CAAX isoprenylation motif present in other GBPs, and localizes to the cytosol by immunofluorescence. Its transcript peaks transiently during erythroid progenitor cell differentiation.","method":"Baculovirus recombinant protein expression, GTPase activity assay, guanine nucleotide-agarose pulldown, immunofluorescence microscopy, Northern blot","journal":"Biochimica et biophysica acta","confidence":"High","confidence_rationale":"Tier 1 — in vitro enzymatic assay with kinetic parameters plus direct localization experiment; single paper but multiple orthogonal methods","pmids":["9659399"],"is_preprint":false},{"year":1994,"finding":"GBP3 was identified as a novel gene on human chromosome 1 with high sequence homology to GBP1 and GBP2, establishing it as a third member of the interferon-inducible GBP gene family.","method":"Genomic library cloning, hybrid rodent-human cell line chromosome mapping, sequence homology analysis","journal":"Gene","confidence":"Medium","confidence_rationale":"Tier 2 — direct chromosomal mapping and structural characterization but no protein-level functional assay","pmids":["7518790"],"is_preprint":false},{"year":2020,"finding":"In IFN-γ-stimulated human cells, GBP3 governs caspase-4 activation on the surface of cytosol-invading Gram-negative bacteria. GBP1 initiates assembly of the GBP signaling platform on bacteria, GBP2 and GBP4 control caspase-4 recruitment, and GBP3 specifically controls caspase-4 activation, which is required for gasdermin-D cleavage, pyroptosis, and IL-18 processing.","method":"Genetic epistasis (GBP knockout/knockdown cells), bacterial infection assays, caspase-4 activation readout, gasdermin-D cleavage assay, IL-18 processing assay, immunofluorescence colocalization","journal":"Nature immunology","confidence":"High","confidence_rationale":"Tier 2 — hierarchical epistasis established with multiple GBP knockouts, multiple orthogonal functional readouts, replicated across labs; 230 citations","pmids":["32541830"],"is_preprint":false},{"year":2017,"finding":"Human GBP1 recruits GBP3 (along with GBP2, GBP4, and GBP6) to the surface of cytosolic Gram-negative bacteria (Shigella flexneri and Burkholderia thailandensis) via GBP1's C-terminal triple-arginine motif, establishing GBP3 as a secondary recruiter dependent on GBP1 for bacterial targeting.","method":"Immunofluorescence colocalization, GBP1 triple-arginine mutant (loss-of-function), bacterial infection assays, siRNA knockdown","journal":"mBio","confidence":"High","confidence_rationale":"Tier 2 — reciprocal colocalization with defined mutagenesis of the recruiter (GBP1), replicated across two bacterial species; 99 citations","pmids":["29233899"],"is_preprint":false},{"year":2019,"finding":"GBP3 is resistant to ubiquitination and proteasomal degradation by the Shigella E3 ligase IpaH9.8. Structural analysis revealed that differences in the Switch II and α3 helix regions of the GTPase domain between GBP1 and GBP3/GBP7 prevent IpaH9.8 LRR domain engagement, making GBP3 refractory to this bacterial immune evasion strategy.","method":"Crystal structure of GBP1–IpaH9.8 LRR complex, structure-guided sequence comparison, ubiquitination and degradation assays","journal":"PLoS pathogens","confidence":"High","confidence_rationale":"Tier 1 — crystal structure with functional validation of substrate selectivity; mechanistic explanation of GBP3 resistance","pmids":["31216343"],"is_preprint":false},{"year":2022,"finding":"Mouse GBP3 (together with mouse GBP1) is specifically required for inflammasome activation during infection with the cytosolic bacterium Francisella novicida. A charged/hydrophobic region within the N-terminal domain of mouse GBP1 and GBP3 directly binds and kills F. novicida and Neisseria meningitidis but not other bacteria or mammalian cells, causing pathogen membrane rupture and release of intracellular content for inflammasome sensing.","method":"GBP1/GBP3 knockout macrophages, bacterial infection assays, direct bacterial killing assay, membrane rupture/permeability assay, domain mapping by mutagenesis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 — clean knockout with defined phenotype plus in vitro killing assay plus domain mutagenesis; multiple orthogonal methods","pmids":["35906252"],"is_preprint":false},{"year":2022,"finding":"Human GBP3 physically interacts with STING via its N-terminal GTPase domain, stabilizing STING protein levels; this interaction promotes expression of p62/SQSTM1, NRF2, and MGMT, thereby enhancing DNA damage repair and conferring temozolomide resistance in glioblastoma cells.","method":"Co-immunoprecipitation, domain deletion constructs, RNA interference knockdown, murine glioblastoma xenograft model, western blot for STING/p62/NRF2/MGMT","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2–3 — Co-IP with domain mapping plus in vivo model; single lab but multiple readouts","pmids":["35780181"],"is_preprint":false},{"year":2017,"finding":"GBP3 promotes glioma cell proliferation by inducing SQSTM1/p62 expression and activating ERK1/2; depletion of SQSTM1 abolished GBP3-driven ERK1/2 phosphorylation and cell growth, and MEK inhibition blocked GBP3-induced proliferation, placing GBP3 upstream of the SQSTM1–ERK1/2 axis.","method":"GBP3 overexpression and siRNA knockdown, ERK1/2 phosphorylation western blot, SQSTM1 siRNA epistasis, MEK inhibitor treatment, in vivo tumor xenograft","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis established by double knockdown/inhibitor experiments; single lab","pmids":["29128363"],"is_preprint":false},{"year":2023,"finding":"GBP3 is not recruited to Francisella novicida (unlike Shigella flexneri) in human macrophages, demonstrating that the repertoire of GBPs assembled on bacteria is pathogen-selective and depends on GBP-intrinsic features and specific bacterial factors. Multiple GBP1 mutagenesis experiments showed that GBP1 targeting to F. novicida requires cooperative engagement of multiple GBP1 domains, whereas targeting to S. flexneri is more permissive.","method":"Immunofluorescence colocalization in human macrophages, GBP1 mutagenesis panel, bacterial infection assays","journal":"Pathogens and disease","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization with mutagenesis; single lab, moderate method depth","pmids":["37012222"],"is_preprint":false},{"year":2023,"finding":"Shigella effector IpaH9.8 promotes shedding of fewer GBPs from Shigella, and in the absence of IpaH9.8, GBP1-dependent LPS release from intracytosolic bacteria is increased, enhancing cytosolic LPS availability for caspase-4 activation. GBP3 (along with other GBPs degraded by IpaH9.8) contributes to caspase-4 activation even in the absence of GBP1.","method":"IpaH9.8 and OspC3 bacterial mutants, GBP knockdown/knockout epithelial cells, cytosolic LPS quantification, caspase-4 activation assay, pyroptosis readout","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 — bacterial genetic and host knockdown epistasis with multiple functional readouts; single lab","pmids":["37014865"],"is_preprint":false},{"year":2017,"finding":"STING (but not cGAS) is required for GBP3 (GBPchr3 locus in mice) expression in macrophages during Brucella abortus infection; GBPchr3 knockout mice are more susceptible to Brucella infection, and siRNA-mediated knockdown reduces IL-1β secretion and caspase-1 activation, placing GBP3 downstream of the STING–IFN axis and upstream of inflammasome activation.","method":"STING/cGAS knockout macrophages, GBPchr3 knockout mice (in vivo infection), siRNA knockdown, IL-1β ELISA, caspase-1 activation assay","journal":"Journal of immunology (Baltimore, Md. : 1950)","confidence":"Medium","confidence_rationale":"Tier 2 — epistasis in vivo and in vitro with defined mechanistic readouts; single lab","pmids":["29203515"],"is_preprint":false},{"year":2025,"finding":"STAT1 directly binds the GBP3 promoter and drives GBP3 transcription in vascular smooth muscle cells stimulated with IFN-γ. GBP3 protein in the cytoplasm physically interacts with STING, forming a STAT1–GBP3–STING positive feedback loop that amplifies inflammation, oxidative stress, and DNA damage in acute aortic dissection.","method":"ChIP-PCR (STAT1 binding to GBP3 promoter), co-immunoprecipitation (GBP3–STING interaction), ATAC-seq/RNA-seq, single-cell RNA sequencing, mouse AAD model with STAT1 inhibitor treatment","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 1–2 — ChIP-PCR and Co-IP establish direct transcriptional regulation and protein interaction; single lab with multiple orthogonal methods","pmids":["40714274"],"is_preprint":false},{"year":2024,"finding":"CRISPR genome-wide knockout screen identified GBP3 as a gene that limits lentiviral vector production; knockout of GBP3 in HEK293T cells increased lentiviral titer, and triple knockout of GBP3, BPIFC, and LDAH achieved ~8.33-fold increase in LV titer, demonstrating GBP3 restricts lentivirus packaging.","method":"CRISPR-Cas9 high-throughput screen, single and multi-gene knockout in HEK293T cells, lentiviral titer quantification","journal":"The CRISPR journal","confidence":"Medium","confidence_rationale":"Tier 2 — direct loss-of-function by CRISPR with quantitative phenotypic readout; single lab","pmids":["39387256"],"is_preprint":false},{"year":2024,"finding":"Gbp3 knockdown in BV2 microglia reduces expression of NLRP3, caspase-1, and GSDMD (pyroptosis pathway proteins) and decreases inflammatory cytokines after oxygen-glucose deprivation; Gbp3 overexpression has the opposite effect, placing Gbp3 as a positive regulator of the NLRP3/GSDMD pyroptotic pathway in ischemic conditions.","method":"siRNA knockdown and plasmid overexpression in BV2 cells, OGD/R model, western blot for NLRP3/GSDMD/caspase-1, ELISA for inflammatory cytokines, rat tMCAO in vivo model","journal":"International immunopharmacology","confidence":"Medium","confidence_rationale":"Tier 2–3 — gain- and loss-of-function with defined pathway readout; single lab, moderate depth","pmids":["39721455"],"is_preprint":false},{"year":2023,"finding":"Gbp3 overexpression in a lupus nephritis cell model inhibits cell proliferation and increases levels of inflammatory factors (IL-1β, TNF-α, IL-8) and pyroptosis-related proteins (GSDMD, caspase-1, NLRP3); siRNA knockdown produces the opposite effects, indicating Gbp3 positively regulates inflammation and pyroptosis in renal cells.","method":"siRNA knockdown and overexpression, CCK-8 proliferation assay, ELISA for cytokines, western blot for pyroptosis proteins, LN mouse model (pristane injection)","journal":"Autoimmunity","confidence":"Low","confidence_rationale":"Tier 3 — gain/loss-of-function with phenotypic readout but no specific molecular binding partner identified; single lab","pmids":["37621179"],"is_preprint":false}],"current_model":"Human/mammalian GBP3 is an IFN-γ-inducible dynamin-like GTPase that, upon cytosolic bacterial invasion, is recruited to the GBP1-nucleated signaling platform on Gram-negative bacteria where it specifically governs caspase-4 activation (leading to gasdermin-D cleavage and pyroptosis); it directly interacts with STING via its GTPase domain to stabilize STING and modulate downstream signaling (including DNA-damage repair and the STAT1–GBP3–STING feedback loop), and its N-terminal domain can directly kill select bacteria (e.g., Francisella novicida) to release ligands for inflammasome sensing, while being intrinsically resistant to Shigella IpaH9.8-mediated ubiquitination due to structural differences in its Switch II/α3 helix regions."},"narrative":{"teleology":[{"year":1994,"claim":"Establishing GBP3 as a member of the interferon-inducible guanylate-binding protein family on human chromosome 1 provided the foundational gene identification needed for functional studies.","evidence":"Genomic library cloning and hybrid cell chromosome mapping","pmids":["7518790"],"confidence":"Medium","gaps":["No protein-level characterization or functional assay performed","Human GBP3 enzymatic activity not measured"]},{"year":1998,"claim":"Demonstrating that mouse GBP3 possesses intrinsic GTPase activity, binds guanine nucleotides, and resides in the cytosol established its biochemical identity as a catalytically active cytosolic GTPase lacking the CAAX prenylation motif present in other family members.","evidence":"Baculovirus-expressed recombinant protein; GTPase kinetic assay, nucleotide-agarose pulldown, immunofluorescence in murine cells","pmids":["9659399"],"confidence":"High","gaps":["Human GBP3 enzymatic parameters not determined","No identification of cellular targets or binding partners","Absence of CAAX motif's functional consequence not tested"]},{"year":2017,"claim":"Showing that GBP1 recruits GBP3 to cytosolic Gram-negative bacteria via GBP1's C-terminal triple-arginine motif established the hierarchical assembly of the GBP defense platform and positioned GBP3 as a downstream effector dependent on GBP1 nucleation.","evidence":"Immunofluorescence colocalization with GBP1 mutants during Shigella and Burkholderia infection in human cells","pmids":["29233899"],"confidence":"High","gaps":["The specific function of GBP3 once recruited was not determined","Direct GBP1–GBP3 protein interaction not biochemically demonstrated"]},{"year":2017,"claim":"Placing GBP3 expression downstream of STING (but not cGAS) during Brucella infection, and showing GBP3-dependent caspase-1 activation and IL-1β secretion, linked GBP3 to the STING–IFN transcriptional axis and inflammasome engagement in vivo.","evidence":"STING/cGAS knockout macrophages, GBPchr3 knockout mice infected with Brucella abortus, siRNA knockdown with IL-1β and caspase-1 readouts","pmids":["29203515"],"confidence":"Medium","gaps":["GBPchr3 locus encodes multiple GBPs; individual gene contributions not fully separated","Mechanism connecting GBP3 to caspase-1 activation not defined"]},{"year":2017,"claim":"Identifying GBP3 as a driver of glioma proliferation upstream of the SQSTM1/p62–ERK1/2 signaling axis revealed a non-canonical, proliferation-promoting role for GBP3 outside of canonical antimicrobial immunity.","evidence":"GBP3 overexpression/knockdown, SQSTM1 epistasis, MEK inhibitor, xenograft model in glioma cells","pmids":["29128363"],"confidence":"Medium","gaps":["Mechanism by which GBP3 induces SQSTM1 expression unknown","Relevance to normal (non-malignant) physiology untested"]},{"year":2019,"claim":"Structural resolution of the GBP1–IpaH9.8 interface revealed that GBP3's Switch II/α3 helix divergence prevents IpaH9.8 engagement, explaining why GBP3 is refractory to Shigella-mediated ubiquitination and proteasomal degradation — a key evasion mechanism sparing GBP3's antimicrobial role.","evidence":"Crystal structure of GBP1–IpaH9.8 LRR complex, structure-guided sequence comparison, ubiquitination and degradation assays","pmids":["31216343"],"confidence":"High","gaps":["Whether other bacterial E3 ligases target GBP3 remains unknown","GBP3's own crystal structure not solved"]},{"year":2020,"claim":"Genetic epistasis across individual GBP knockouts demonstrated that GBP3 specifically controls caspase-4 activation (not recruitment) on cytosolic bacteria, placing it as the rate-limiting step for gasdermin-D cleavage, pyroptosis, and IL-18 processing within the GBP platform.","evidence":"Systematic GBP knockout/knockdown in human epithelial cells, Gram-negative bacterial infection, caspase-4 activation and GSDMD cleavage assays","pmids":["32541830"],"confidence":"High","gaps":["The biochemical mechanism by which GBP3 activates caspase-4 (direct interaction, conformational change, or cofactor) is unknown","Whether GBP3 GTPase activity is required for caspase-4 activation not shown"]},{"year":2022,"claim":"Discovery that the N-terminal domain of mouse GBP3 (and GBP1) directly kills Francisella novicida by rupturing pathogen membranes established a bactericidal effector function for GBP3 that liberates microbial ligands for inflammasome sensing.","evidence":"GBP1/GBP3 knockout macrophages, direct bacterial killing and membrane permeability assays, domain-mapping mutagenesis","pmids":["35906252"],"confidence":"High","gaps":["Killing is selective for certain bacteria; determinants of susceptibility unknown","Whether human GBP3 possesses the same direct bactericidal activity not tested"]},{"year":2022,"claim":"Demonstrating that GBP3 physically binds STING through its GTPase domain and stabilizes STING protein revealed a direct molecular link between GBP3 and the cGAS–STING axis, with downstream effects on DNA-damage repair (MGMT, NRF2) and temozolomide resistance in glioblastoma.","evidence":"Co-immunoprecipitation with domain deletions, siRNA knockdown, xenograft tumor model, western blot for STING/p62/NRF2/MGMT","pmids":["35780181"],"confidence":"Medium","gaps":["Reciprocal Co-IP from endogenous STING not shown","Whether GBP3 GTPase activity modulates STING binding/stabilization unknown","Structural basis of GBP3–STING interaction not resolved"]},{"year":2023,"claim":"Showing that GBP3 is not recruited to Francisella novicida in human macrophages (unlike Shigella) demonstrated that GBP platform composition is pathogen-selective and depends on bacterial surface determinants, refining the model of GBP3 targeting.","evidence":"Immunofluorescence colocalization with GBP1 mutagenesis panel in human macrophages infected with F. novicida versus S. flexneri","pmids":["37012222"],"confidence":"Medium","gaps":["Bacterial surface factor(s) that determine GBP3 recruitment specificity not identified","Whether human GBP3 exerts bactericidal activity against F. novicida independent of recruitment unknown"]},{"year":2023,"claim":"Epistasis between IpaH9.8 and GBP knockouts showed that GBP3 contributes to caspase-4 activation and LPS sensing even when GBP1 is absent, indicating partially redundant GBP-mediated pathways for cytosolic LPS detection.","evidence":"IpaH9.8/OspC3 bacterial mutants, GBP knockdown/knockout in epithelial cells, cytosolic LPS quantification, caspase-4 activation","pmids":["37014865"],"confidence":"Medium","gaps":["Quantitative contribution of GBP3 alone to LPS release not isolated","Mechanism by which GBP3 promotes LPS access without GBP1 unknown"]},{"year":2025,"claim":"Identification of STAT1 directly binding the GBP3 promoter and GBP3 reciprocally stabilizing STING established a STAT1→GBP3→STING positive feedback circuit amplifying inflammation, oxidative stress, and DNA damage in vascular smooth muscle cells during aortic dissection.","evidence":"ChIP-PCR for STAT1 at GBP3 promoter, Co-IP for GBP3–STING, ATAC-seq/RNA-seq, scRNA-seq, mouse aortic dissection model with STAT1 inhibitor","pmids":["40714274"],"confidence":"Medium","gaps":["Whether the feedback loop operates in immune cells beyond vascular smooth muscle not tested","Quantitative contribution of GBP3 versus other STAT1 targets to STING stabilization unclear"]},{"year":null,"claim":"The biochemical mechanism by which GBP3 activates caspase-4 — whether through direct binding, GTPase-dependent conformational change, or facilitated LPS presentation — remains the central unresolved question for GBP3 innate immune function.","evidence":"","pmids":[],"confidence":"High","gaps":["No direct GBP3–caspase-4 binding demonstrated","GBP3 crystal structure not solved","Whether GBP3 GTPase catalytic activity is required for any of its effector functions untested"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003924","term_label":"GTPase activity","supporting_discovery_ids":[0]},{"term_id":"GO:0016787","term_label":"hydrolase activity","supporting_discovery_ids":[0]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,13]}],"localization":[{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[0,6,11]}],"pathway":[{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[2,3,5,10]},{"term_id":"R-HSA-5357801","term_label":"Programmed Cell Death","supporting_discovery_ids":[2,13,14]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[6,7,11]}],"complexes":[],"partners":["GBP1","STING","CASP4","SQSTM1"],"other_free_text":[]},"mechanistic_narrative":"GBP3 is an interferon-γ–inducible dynamin-superfamily GTPase that functions as a critical effector in cell-autonomous innate immunity against cytosolic Gram-negative bacteria and as a positive regulator of inflammasome-driven pyroptosis. Within the hierarchical GBP signaling platform assembled on intracellular bacteria, GBP3 is recruited in a GBP1-dependent manner and specifically governs caspase-4 activation, leading to gasdermin-D cleavage, pyroptosis, and IL-18 processing; its GTPase domain structural divergence in the Switch II/α3 helix renders it resistant to the Shigella E3 ligase IpaH9.8, preserving its antimicrobial function [PMID:32541830, PMID:29233899, PMID:31216343]. In mouse macrophages, GBP3 directly kills select bacteria such as Francisella novicida through an N-terminal bactericidal domain, releasing pathogen contents for inflammasome sensing, and is itself transcriptionally dependent on the STING–IFN axis [PMID:35906252, PMID:29203515]. GBP3 also physically interacts with STING via its GTPase domain to stabilize STING protein, participating in a STAT1–GBP3–STING positive feedback loop that amplifies inflammatory and DNA-damage signaling in contexts including glioblastoma chemoresistance and vascular inflammation [PMID:35780181, PMID:40714274]."},"prefetch_data":{"uniprot":{"accession":"Q9H0R5","full_name":"Guanylate-binding protein 3","aliases":["GTP-binding protein 3","GBP-3","Guanine nucleotide-binding protein 3"],"length_aa":595,"mass_kda":68.1,"function":"Interferon (IFN)-inducible GTPase that plays important roles in innate immunity against a diverse range of bacterial, viral and protozoan pathogens (PubMed:22106366). Hydrolyzes GTP very efficiently; GDP rather than GMP is the major reaction product (By similarity). Following infection, recruited to the pathogen-containing vacuoles or vacuole-escaped bacteria and acts as a positive regulator of inflammasome assembly by promoting the release of inflammasome ligands from bacteria (By similarity). Acts by promoting lysis of pathogen-containing vacuoles, releasing pathogens into the cytosol (By similarity). Following pathogen release in the cytosol, promotes recruitment of proteins that mediate bacterial cytolysis: this liberates ligands that are detected by inflammasomes, such as lipopolysaccharide (LPS) that activates the non-canonical CASP4/CASP11 inflammasome or double-stranded DNA (dsDNA) that activates the AIM2 inflammasome (By similarity). Exhibits antiviral activity against influenza virus (PubMed:22106366) Shows the most prominent antiviral activity in epithelial cells","subcellular_location":"Cytoplasm; Cytoplasm, perinuclear region; Golgi apparatus membrane","url":"https://www.uniprot.org/uniprotkb/Q9H0R5/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/GBP3","classification":"Not Classified","n_dependent_lines":6,"n_total_lines":1208,"dependency_fraction":0.004966887417218543},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/GBP3","total_profiled":1310},"omim":[{"mim_id":"600413","title":"GUANYLATE-BINDING PROTEIN 3; GBP3","url":"https://www.omim.org/entry/600413"},{"mim_id":"600412","title":"GUANYLATE-BINDING PROTEIN 2, INTERFERON-INDUCIBLE; GBP2","url":"https://www.omim.org/entry/600412"},{"mim_id":"600411","title":"GUANYLATE-BINDING PROTEIN 1, INTERFERON-INDUCIBLE, 67-KD; GBP1","url":"https://www.omim.org/entry/600411"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"intestine","ntpm":74.1}],"url":"https://www.proteinatlas.org/search/GBP3"},"hgnc":{"alias_symbol":["FLJ10961"],"prev_symbol":[]},"alphafold":{"accession":"Q8WXF7","domains":[{"cath_id":"3.40.50.300","chopping":"33-344","consensus_level":"medium","plddt":94.8913,"start":33,"end":344},{"cath_id":"1.20.58.420","chopping":"345-532","consensus_level":"medium","plddt":85.7354,"start":345,"end":532}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8WXF7","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8WXF7-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8WXF7-F1-predicted_aligned_error_v6.png","plddt_mean":86.56},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=GBP3","jax_strain_url":"https://www.jax.org/strain/search?query=GBP3"},"sequence":{"accession":"Q8WXF7","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8WXF7.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8WXF7/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8WXF7"}},"corpus_meta":[{"pmid":"32541830","id":"PMC_32541830","title":"Guanylate-binding 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Its transcript peaks transiently during erythroid progenitor cell differentiation.\",\n      \"method\": \"Baculovirus recombinant protein expression, GTPase activity assay, guanine nucleotide-agarose pulldown, immunofluorescence microscopy, Northern blot\",\n      \"journal\": \"Biochimica et biophysica acta\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro enzymatic assay with kinetic parameters plus direct localization experiment; single paper but multiple orthogonal methods\",\n      \"pmids\": [\"9659399\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"GBP3 was identified as a novel gene on human chromosome 1 with high sequence homology to GBP1 and GBP2, establishing it as a third member of the interferon-inducible GBP gene family.\",\n      \"method\": \"Genomic library cloning, hybrid rodent-human cell line chromosome mapping, sequence homology analysis\",\n      \"journal\": \"Gene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct chromosomal mapping and structural characterization but no protein-level functional assay\",\n      \"pmids\": [\"7518790\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In IFN-γ-stimulated human cells, GBP3 governs caspase-4 activation on the surface of cytosol-invading Gram-negative bacteria. GBP1 initiates assembly of the GBP signaling platform on bacteria, GBP2 and GBP4 control caspase-4 recruitment, and GBP3 specifically controls caspase-4 activation, which is required for gasdermin-D cleavage, pyroptosis, and IL-18 processing.\",\n      \"method\": \"Genetic epistasis (GBP knockout/knockdown cells), bacterial infection assays, caspase-4 activation readout, gasdermin-D cleavage assay, IL-18 processing assay, immunofluorescence colocalization\",\n      \"journal\": \"Nature immunology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — hierarchical epistasis established with multiple GBP knockouts, multiple orthogonal functional readouts, replicated across labs; 230 citations\",\n      \"pmids\": [\"32541830\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Human GBP1 recruits GBP3 (along with GBP2, GBP4, and GBP6) to the surface of cytosolic Gram-negative bacteria (Shigella flexneri and Burkholderia thailandensis) via GBP1's C-terminal triple-arginine motif, establishing GBP3 as a secondary recruiter dependent on GBP1 for bacterial targeting.\",\n      \"method\": \"Immunofluorescence colocalization, GBP1 triple-arginine mutant (loss-of-function), bacterial infection assays, siRNA knockdown\",\n      \"journal\": \"mBio\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal colocalization with defined mutagenesis of the recruiter (GBP1), replicated across two bacterial species; 99 citations\",\n      \"pmids\": [\"29233899\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"GBP3 is resistant to ubiquitination and proteasomal degradation by the Shigella E3 ligase IpaH9.8. Structural analysis revealed that differences in the Switch II and α3 helix regions of the GTPase domain between GBP1 and GBP3/GBP7 prevent IpaH9.8 LRR domain engagement, making GBP3 refractory to this bacterial immune evasion strategy.\",\n      \"method\": \"Crystal structure of GBP1–IpaH9.8 LRR complex, structure-guided sequence comparison, ubiquitination and degradation assays\",\n      \"journal\": \"PLoS pathogens\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure with functional validation of substrate selectivity; mechanistic explanation of GBP3 resistance\",\n      \"pmids\": [\"31216343\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Mouse GBP3 (together with mouse GBP1) is specifically required for inflammasome activation during infection with the cytosolic bacterium Francisella novicida. A charged/hydrophobic region within the N-terminal domain of mouse GBP1 and GBP3 directly binds and kills F. novicida and Neisseria meningitidis but not other bacteria or mammalian cells, causing pathogen membrane rupture and release of intracellular content for inflammasome sensing.\",\n      \"method\": \"GBP1/GBP3 knockout macrophages, bacterial infection assays, direct bacterial killing assay, membrane rupture/permeability assay, domain mapping by mutagenesis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — clean knockout with defined phenotype plus in vitro killing assay plus domain mutagenesis; multiple orthogonal methods\",\n      \"pmids\": [\"35906252\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Human GBP3 physically interacts with STING via its N-terminal GTPase domain, stabilizing STING protein levels; this interaction promotes expression of p62/SQSTM1, NRF2, and MGMT, thereby enhancing DNA damage repair and conferring temozolomide resistance in glioblastoma cells.\",\n      \"method\": \"Co-immunoprecipitation, domain deletion constructs, RNA interference knockdown, murine glioblastoma xenograft model, western blot for STING/p62/NRF2/MGMT\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — Co-IP with domain mapping plus in vivo model; single lab but multiple readouts\",\n      \"pmids\": [\"35780181\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"GBP3 promotes glioma cell proliferation by inducing SQSTM1/p62 expression and activating ERK1/2; depletion of SQSTM1 abolished GBP3-driven ERK1/2 phosphorylation and cell growth, and MEK inhibition blocked GBP3-induced proliferation, placing GBP3 upstream of the SQSTM1–ERK1/2 axis.\",\n      \"method\": \"GBP3 overexpression and siRNA knockdown, ERK1/2 phosphorylation western blot, SQSTM1 siRNA epistasis, MEK inhibitor treatment, in vivo tumor xenograft\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis established by double knockdown/inhibitor experiments; single lab\",\n      \"pmids\": [\"29128363\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"GBP3 is not recruited to Francisella novicida (unlike Shigella flexneri) in human macrophages, demonstrating that the repertoire of GBPs assembled on bacteria is pathogen-selective and depends on GBP-intrinsic features and specific bacterial factors. Multiple GBP1 mutagenesis experiments showed that GBP1 targeting to F. novicida requires cooperative engagement of multiple GBP1 domains, whereas targeting to S. flexneri is more permissive.\",\n      \"method\": \"Immunofluorescence colocalization in human macrophages, GBP1 mutagenesis panel, bacterial infection assays\",\n      \"journal\": \"Pathogens and disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization with mutagenesis; single lab, moderate method depth\",\n      \"pmids\": [\"37012222\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Shigella effector IpaH9.8 promotes shedding of fewer GBPs from Shigella, and in the absence of IpaH9.8, GBP1-dependent LPS release from intracytosolic bacteria is increased, enhancing cytosolic LPS availability for caspase-4 activation. GBP3 (along with other GBPs degraded by IpaH9.8) contributes to caspase-4 activation even in the absence of GBP1.\",\n      \"method\": \"IpaH9.8 and OspC3 bacterial mutants, GBP knockdown/knockout epithelial cells, cytosolic LPS quantification, caspase-4 activation assay, pyroptosis readout\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — bacterial genetic and host knockdown epistasis with multiple functional readouts; single lab\",\n      \"pmids\": [\"37014865\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"STING (but not cGAS) is required for GBP3 (GBPchr3 locus in mice) expression in macrophages during Brucella abortus infection; GBPchr3 knockout mice are more susceptible to Brucella infection, and siRNA-mediated knockdown reduces IL-1β secretion and caspase-1 activation, placing GBP3 downstream of the STING–IFN axis and upstream of inflammasome activation.\",\n      \"method\": \"STING/cGAS knockout macrophages, GBPchr3 knockout mice (in vivo infection), siRNA knockdown, IL-1β ELISA, caspase-1 activation assay\",\n      \"journal\": \"Journal of immunology (Baltimore, Md. : 1950)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — epistasis in vivo and in vitro with defined mechanistic readouts; single lab\",\n      \"pmids\": [\"29203515\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"STAT1 directly binds the GBP3 promoter and drives GBP3 transcription in vascular smooth muscle cells stimulated with IFN-γ. GBP3 protein in the cytoplasm physically interacts with STING, forming a STAT1–GBP3–STING positive feedback loop that amplifies inflammation, oxidative stress, and DNA damage in acute aortic dissection.\",\n      \"method\": \"ChIP-PCR (STAT1 binding to GBP3 promoter), co-immunoprecipitation (GBP3–STING interaction), ATAC-seq/RNA-seq, single-cell RNA sequencing, mouse AAD model with STAT1 inhibitor treatment\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 — ChIP-PCR and Co-IP establish direct transcriptional regulation and protein interaction; single lab with multiple orthogonal methods\",\n      \"pmids\": [\"40714274\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CRISPR genome-wide knockout screen identified GBP3 as a gene that limits lentiviral vector production; knockout of GBP3 in HEK293T cells increased lentiviral titer, and triple knockout of GBP3, BPIFC, and LDAH achieved ~8.33-fold increase in LV titer, demonstrating GBP3 restricts lentivirus packaging.\",\n      \"method\": \"CRISPR-Cas9 high-throughput screen, single and multi-gene knockout in HEK293T cells, lentiviral titer quantification\",\n      \"journal\": \"The CRISPR journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct loss-of-function by CRISPR with quantitative phenotypic readout; single lab\",\n      \"pmids\": [\"39387256\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Gbp3 knockdown in BV2 microglia reduces expression of NLRP3, caspase-1, and GSDMD (pyroptosis pathway proteins) and decreases inflammatory cytokines after oxygen-glucose deprivation; Gbp3 overexpression has the opposite effect, placing Gbp3 as a positive regulator of the NLRP3/GSDMD pyroptotic pathway in ischemic conditions.\",\n      \"method\": \"siRNA knockdown and plasmid overexpression in BV2 cells, OGD/R model, western blot for NLRP3/GSDMD/caspase-1, ELISA for inflammatory cytokines, rat tMCAO in vivo model\",\n      \"journal\": \"International immunopharmacology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — gain- and loss-of-function with defined pathway readout; single lab, moderate depth\",\n      \"pmids\": [\"39721455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Gbp3 overexpression in a lupus nephritis cell model inhibits cell proliferation and increases levels of inflammatory factors (IL-1β, TNF-α, IL-8) and pyroptosis-related proteins (GSDMD, caspase-1, NLRP3); siRNA knockdown produces the opposite effects, indicating Gbp3 positively regulates inflammation and pyroptosis in renal cells.\",\n      \"method\": \"siRNA knockdown and overexpression, CCK-8 proliferation assay, ELISA for cytokines, western blot for pyroptosis proteins, LN mouse model (pristane injection)\",\n      \"journal\": \"Autoimmunity\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — gain/loss-of-function with phenotypic readout but no specific molecular binding partner identified; single lab\",\n      \"pmids\": [\"37621179\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"Human/mammalian GBP3 is an IFN-γ-inducible dynamin-like GTPase that, upon cytosolic bacterial invasion, is recruited to the GBP1-nucleated signaling platform on Gram-negative bacteria where it specifically governs caspase-4 activation (leading to gasdermin-D cleavage and pyroptosis); it directly interacts with STING via its GTPase domain to stabilize STING and modulate downstream signaling (including DNA-damage repair and the STAT1–GBP3–STING feedback loop), and its N-terminal domain can directly kill select bacteria (e.g., Francisella novicida) to release ligands for inflammasome sensing, while being intrinsically resistant to Shigella IpaH9.8-mediated ubiquitination due to structural differences in its Switch II/α3 helix regions.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"GBP3 is an interferon-γ–inducible dynamin-superfamily GTPase that functions as a critical effector in cell-autonomous innate immunity against cytosolic Gram-negative bacteria and as a positive regulator of inflammasome-driven pyroptosis. Within the hierarchical GBP signaling platform assembled on intracellular bacteria, GBP3 is recruited in a GBP1-dependent manner and specifically governs caspase-4 activation, leading to gasdermin-D cleavage, pyroptosis, and IL-18 processing; its GTPase domain structural divergence in the Switch II/α3 helix renders it resistant to the Shigella E3 ligase IpaH9.8, preserving its antimicrobial function [PMID:32541830, PMID:29233899, PMID:31216343]. In mouse macrophages, GBP3 directly kills select bacteria such as Francisella novicida through an N-terminal bactericidal domain, releasing pathogen contents for inflammasome sensing, and is itself transcriptionally dependent on the STING–IFN axis [PMID:35906252, PMID:29203515]. GBP3 also physically interacts with STING via its GTPase domain to stabilize STING protein, participating in a STAT1–GBP3–STING positive feedback loop that amplifies inflammatory and DNA-damage signaling in contexts including glioblastoma chemoresistance and vascular inflammation [PMID:35780181, PMID:40714274].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Establishing GBP3 as a member of the interferon-inducible guanylate-binding protein family on human chromosome 1 provided the foundational gene identification needed for functional studies.\",\n      \"evidence\": \"Genomic library cloning and hybrid cell chromosome mapping\",\n      \"pmids\": [\"7518790\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No protein-level characterization or functional assay performed\", \"Human GBP3 enzymatic activity not measured\"]\n    },\n    {\n      \"year\": 1998,\n      \"claim\": \"Demonstrating that mouse GBP3 possesses intrinsic GTPase activity, binds guanine nucleotides, and resides in the cytosol established its biochemical identity as a catalytically active cytosolic GTPase lacking the CAAX prenylation motif present in other family members.\",\n      \"evidence\": \"Baculovirus-expressed recombinant protein; GTPase kinetic assay, nucleotide-agarose pulldown, immunofluorescence in murine cells\",\n      \"pmids\": [\"9659399\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Human GBP3 enzymatic parameters not determined\", \"No identification of cellular targets or binding partners\", \"Absence of CAAX motif's functional consequence not tested\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Showing that GBP1 recruits GBP3 to cytosolic Gram-negative bacteria via GBP1's C-terminal triple-arginine motif established the hierarchical assembly of the GBP defense platform and positioned GBP3 as a downstream effector dependent on GBP1 nucleation.\",\n      \"evidence\": \"Immunofluorescence colocalization with GBP1 mutants during Shigella and Burkholderia infection in human cells\",\n      \"pmids\": [\"29233899\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The specific function of GBP3 once recruited was not determined\", \"Direct GBP1–GBP3 protein interaction not biochemically demonstrated\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Placing GBP3 expression downstream of STING (but not cGAS) during Brucella infection, and showing GBP3-dependent caspase-1 activation and IL-1β secretion, linked GBP3 to the STING–IFN transcriptional axis and inflammasome engagement in vivo.\",\n      \"evidence\": \"STING/cGAS knockout macrophages, GBPchr3 knockout mice infected with Brucella abortus, siRNA knockdown with IL-1β and caspase-1 readouts\",\n      \"pmids\": [\"29203515\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"GBPchr3 locus encodes multiple GBPs; individual gene contributions not fully separated\", \"Mechanism connecting GBP3 to caspase-1 activation not defined\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identifying GBP3 as a driver of glioma proliferation upstream of the SQSTM1/p62–ERK1/2 signaling axis revealed a non-canonical, proliferation-promoting role for GBP3 outside of canonical antimicrobial immunity.\",\n      \"evidence\": \"GBP3 overexpression/knockdown, SQSTM1 epistasis, MEK inhibitor, xenograft model in glioma cells\",\n      \"pmids\": [\"29128363\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which GBP3 induces SQSTM1 expression unknown\", \"Relevance to normal (non-malignant) physiology untested\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Structural resolution of the GBP1–IpaH9.8 interface revealed that GBP3's Switch II/α3 helix divergence prevents IpaH9.8 engagement, explaining why GBP3 is refractory to Shigella-mediated ubiquitination and proteasomal degradation — a key evasion mechanism sparing GBP3's antimicrobial role.\",\n      \"evidence\": \"Crystal structure of GBP1–IpaH9.8 LRR complex, structure-guided sequence comparison, ubiquitination and degradation assays\",\n      \"pmids\": [\"31216343\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other bacterial E3 ligases target GBP3 remains unknown\", \"GBP3's own crystal structure not solved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Genetic epistasis across individual GBP knockouts demonstrated that GBP3 specifically controls caspase-4 activation (not recruitment) on cytosolic bacteria, placing it as the rate-limiting step for gasdermin-D cleavage, pyroptosis, and IL-18 processing within the GBP platform.\",\n      \"evidence\": \"Systematic GBP knockout/knockdown in human epithelial cells, Gram-negative bacterial infection, caspase-4 activation and GSDMD cleavage assays\",\n      \"pmids\": [\"32541830\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"The biochemical mechanism by which GBP3 activates caspase-4 (direct interaction, conformational change, or cofactor) is unknown\", \"Whether GBP3 GTPase activity is required for caspase-4 activation not shown\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Discovery that the N-terminal domain of mouse GBP3 (and GBP1) directly kills Francisella novicida by rupturing pathogen membranes established a bactericidal effector function for GBP3 that liberates microbial ligands for inflammasome sensing.\",\n      \"evidence\": \"GBP1/GBP3 knockout macrophages, direct bacterial killing and membrane permeability assays, domain-mapping mutagenesis\",\n      \"pmids\": [\"35906252\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Killing is selective for certain bacteria; determinants of susceptibility unknown\", \"Whether human GBP3 possesses the same direct bactericidal activity not tested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Demonstrating that GBP3 physically binds STING through its GTPase domain and stabilizes STING protein revealed a direct molecular link between GBP3 and the cGAS–STING axis, with downstream effects on DNA-damage repair (MGMT, NRF2) and temozolomide resistance in glioblastoma.\",\n      \"evidence\": \"Co-immunoprecipitation with domain deletions, siRNA knockdown, xenograft tumor model, western blot for STING/p62/NRF2/MGMT\",\n      \"pmids\": [\"35780181\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reciprocal Co-IP from endogenous STING not shown\", \"Whether GBP3 GTPase activity modulates STING binding/stabilization unknown\", \"Structural basis of GBP3–STING interaction not resolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Showing that GBP3 is not recruited to Francisella novicida in human macrophages (unlike Shigella) demonstrated that GBP platform composition is pathogen-selective and depends on bacterial surface determinants, refining the model of GBP3 targeting.\",\n      \"evidence\": \"Immunofluorescence colocalization with GBP1 mutagenesis panel in human macrophages infected with F. novicida versus S. flexneri\",\n      \"pmids\": [\"37012222\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Bacterial surface factor(s) that determine GBP3 recruitment specificity not identified\", \"Whether human GBP3 exerts bactericidal activity against F. novicida independent of recruitment unknown\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Epistasis between IpaH9.8 and GBP knockouts showed that GBP3 contributes to caspase-4 activation and LPS sensing even when GBP1 is absent, indicating partially redundant GBP-mediated pathways for cytosolic LPS detection.\",\n      \"evidence\": \"IpaH9.8/OspC3 bacterial mutants, GBP knockdown/knockout in epithelial cells, cytosolic LPS quantification, caspase-4 activation\",\n      \"pmids\": [\"37014865\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Quantitative contribution of GBP3 alone to LPS release not isolated\", \"Mechanism by which GBP3 promotes LPS access without GBP1 unknown\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identification of STAT1 directly binding the GBP3 promoter and GBP3 reciprocally stabilizing STING established a STAT1→GBP3→STING positive feedback circuit amplifying inflammation, oxidative stress, and DNA damage in vascular smooth muscle cells during aortic dissection.\",\n      \"evidence\": \"ChIP-PCR for STAT1 at GBP3 promoter, Co-IP for GBP3–STING, ATAC-seq/RNA-seq, scRNA-seq, mouse aortic dissection model with STAT1 inhibitor\",\n      \"pmids\": [\"40714274\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the feedback loop operates in immune cells beyond vascular smooth muscle not tested\", \"Quantitative contribution of GBP3 versus other STAT1 targets to STING stabilization unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The biochemical mechanism by which GBP3 activates caspase-4 — whether through direct binding, GTPase-dependent conformational change, or facilitated LPS presentation — remains the central unresolved question for GBP3 innate immune function.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No direct GBP3–caspase-4 binding demonstrated\", \"GBP3 crystal structure not solved\", \"Whether GBP3 GTPase catalytic activity is required for any of its effector functions untested\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003924\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0016787\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 13]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [0, 6, 11]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [2, 3, 5, 10]},\n      {\"term_id\": \"R-HSA-5357801\", \"supporting_discovery_ids\": [2, 13, 14]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [6, 7, 11]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\n      \"GBP1\",\n      \"STING\",\n      \"CASP4\",\n      \"SQSTM1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}