{"gene":"APBA3","run_date":"2026-06-09T22:02:43","timeline":{"discoveries":[{"year":1999,"finding":"APBA3/X11L2 PID/PTB domain directly interacts with the intracellular domain of APP (amyloid precursor protein), as demonstrated by GST binding assay in vitro and confirmed by co-immunoprecipitation from cells overexpressing APP and HA-tagged X11L2.","method":"GST pull-down assay, co-immunoprecipitation","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP plus in vitro GST pull-down, single lab, two orthogonal methods","pmids":["10049767"],"is_preprint":false},{"year":2009,"finding":"Mint3/APBA3 binds directly to FIH-1 (factor inhibiting HIF-1) and inhibits FIH-1's ability to hydroxylate HIF-1α Asn803, thereby preventing FIH-1-mediated suppression of HIF-1 transcriptional activity in an oxygen-independent manner. In macrophages, this mechanism sustains HIF-1 activity during normoxia and supports glycolytic ATP production. Knockdown of Mint3 redistributes FIH-1 from the perinuclear region to the cytoplasm and decreases glycolysis and ATP production.","method":"Purified protein binding assay (in vitro), HIF-1 reporter assay, siRNA knockdown, immunofluorescence co-localization, biochemical fractionation","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro reconstituted inhibition assay plus reporter assay plus KD phenotype in macrophages, multiple orthogonal methods in one study","pmids":["19726677"],"is_preprint":false},{"year":2007,"finding":"Mint3 is specifically enriched in APP-containing vesicles purified from neuroblastoma cells, and depletion of Mint3 by siRNA redirects APP export from the basolateral/trans-Golgi route to the endosomal/lysosomal route; Mint3 overexpression decreases and siRNA knockdown increases Aβ(1-40) secretion, identifying Mint3 as a critical determinant of post-Golgi APP trafficking.","method":"Vesicle purification from cells, siRNA knockdown, subcellular localization/trafficking assay, Aβ ELISA","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — vesicle purification plus siRNA KD plus functional Aβ measurement, multiple orthogonal methods","pmids":["17959829"],"is_preprint":false},{"year":2004,"finding":"The PDZ domains of Mint-3 bind the carboxyl EWV motif of MT5-MMP (identified by yeast two-hybrid screening), and this interaction mediates retrieval of internalized MT5-MMP to the plasma membrane; deletion of EWV impairs recycling without affecting internalization, and siRNA-mediated knockdown of Mint-3 decreases MT5-MMP surface activity.","method":"Yeast two-hybrid, siRNA knockdown, cell surface activity assay, deletion mutagenesis","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — yeast two-hybrid interaction mapped to specific motif, mutagenesis of EWV, siRNA functional validation, single lab but multiple orthogonal methods","pmids":["14990567"],"is_preprint":false},{"year":2005,"finding":"Rab6A GTPase interacts with Mint3 in a GTP-dependent manner, requiring the complete PTB domain of Mint3. Rab6A, Mint3, and APP co-localize at Golgi membranes in HeLa cells, suggesting Mint3 links Rab6A to APP trafficking.","method":"Yeast two-hybrid, confocal microscopy co-localization, density gradient centrifugation, GTP-dependency assay","journal":"Biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — yeast two-hybrid plus co-localization plus fractionation, single lab, no reciprocal Co-IP in mammalian cells","pmids":["16207088"],"is_preprint":false},{"year":2008,"finding":"Mint3 PTB domain interacts with the acidic peptide signal of Furin in HeLa cells (shown by co-immunoprecipitation and immunofluorescence), and Mint3 knockdown by RNAi disrupts TGN-specific localization of Furin, redistributing it to endosomes.","method":"Co-immunoprecipitation, immunofluorescence, siRNA knockdown, domain mapping/mutagenesis","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal Co-IP plus siRNA KD with clear localization phenotype plus mutagenesis, single lab","pmids":["18544638"],"is_preprint":false},{"year":2013,"finding":"Mint3 is recruited to the Golgi by the YENPTY motif of APP specifically via Tyr-682, and this recruitment is required for APP export from the Golgi; after leaving the Golgi, APP is directed to LAMP1+ structures as the proximal destination.","method":"Site-directed mutagenesis of APP sorting motifs, subcellular localization assay, adaptor recruitment assay","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — systematic mutagenesis of all four APP sorting motifs, specific residue identified, single lab","pmids":["23965993"],"is_preprint":false},{"year":2011,"finding":"Genetic deletion of Apba3 in mice reduces ATP production in macrophages to ~60% of wild-type, decreases glycolysis, cytokine production, and motility, and confers resistance to LPS-induced septic shock; myeloid-specific deletion recapitulates the septic shock resistance, establishing a cell-type-specific role for APBA3 in macrophage energy metabolism via the FIH-1–HIF-1 pathway.","method":"Apba3 knockout mice, myeloid-specific conditional knockout, ATP measurement, cytokine assay, LPS-induced septic shock model","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — germline and conditional KO mice with multiple functional readouts, mechanistic pathway confirmed in vivo","pmids":["21778228"],"is_preprint":false},{"year":2007,"finding":"X11L2/APBA3 shuttles between the cytoplasm and nucleus via a nuclear export signal (NES) in its N-terminus; mutation of the NES causes nuclear accumulation, and X11L2 tethered near a promoter (via Gal4-DBD fusion) displays transcriptional activator activity, which is attenuated by preventing nucleo-cytoplasmic shuttling.","method":"Leptomycin B treatment, FLIP (fluorescence loss in photobleaching), NES mutagenesis, Gal4-DBD transcription reporter assay, nuclear fractionation","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — FLIP live imaging plus NES mutagenesis plus functional reporter assay, single lab, multiple orthogonal methods","pmids":["18201694"],"is_preprint":false},{"year":2004,"finding":"The C-terminal PDZ-binding motif of Bcr (breakpoint cluster region protein) specifically binds to the PDZ domains of Mint3, localizing Bcr to the Golgi compartment where Mint3 resides.","method":"PDZ domain binding assay, co-localization immunofluorescence","journal":"Journal of cell science","confidence":"Low","confidence_rationale":"Tier 3 / Weak — PDZ binding assay and co-localization, single lab, single method type, limited mechanistic follow-up for Mint3 specifically","pmids":["15494376"],"is_preprint":false},{"year":2010,"finding":"The PTB domain of Mint3 (Mint3Δ6) is sufficient to interact with constitutively active (GTP-loaded) Rab6A in living mammalian cells, as confirmed by FACS-based FRET analysis; a mutant lacking part of the PTB domain (Mint3Δ4) fails to interact.","method":"FACS-based FRET analysis in live cells, GST pull-down","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — FRET in live cells plus GST pull-down with domain-mapping mutants, single lab, two orthogonal methods","pmids":["20447381"],"is_preprint":false},{"year":2014,"finding":"In nucleus pulposus (NP) cells, overexpressed FIH-1 suppresses HIF-1 activity, and co-transfection of full-length Mint3 or the Mint3 N-terminus abrogates this suppression under both normoxia and hypoxia. However, endogenous FIH-1 silencing does not significantly change classical HIF-1 target gene transcripts in NP cells, indicating the FIH-1–Mint3 axis does not control endogenous HIF-1 transcriptional activity in this cell type.","method":"HIF-1 reporter assay, co-transfection, nuclear import/export inhibitors, microarray after FIH-1 silencing","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reporter assay plus domain mapping plus microarray, single lab; notable that this is a negative result for NP cell physiology","pmids":["24867948"],"is_preprint":false},{"year":2016,"finding":"Mint3 promotes K63-linked polyubiquitination of TRAF3, thereby enhancing IRF3 activation and IFN-β production downstream of TLR3/4 and RIG-I signaling in macrophages; Mint3 deficiency greatly attenuates antiviral immune responses and increases viral replication.","method":"Mint3 KO/knockdown in macrophages, ubiquitination assay (K63-linked), IRF3 activation assay, IFN-β reporter/ELISA, viral replication assay","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mice and KD with mechanistic ubiquitination assay, single lab","pmids":["27698125"],"is_preprint":false},{"year":2016,"finding":"In macrophages, Mint3 depletion attenuates NF-κB signaling (by increasing IκBα) and activates AMPK, reducing cytokine/chemokine production in response to influenza virus; Mint3-deficient mice show reduced inflammatory cytokines and neutrophil infiltration during influenza pneumonia without altering viral burden.","method":"Mint3 KO mice, siRNA knockdown, NF-κB pathway analysis (IκBα levels), AMPK activation assay, cytokine ELISA, neutrophil infiltration histology","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — KO mouse model plus in vitro pathway analysis with two distinct signaling readouts, single lab","pmids":["27883071"],"is_preprint":false},{"year":2017,"finding":"Mint3/APBA3 in inflammatory monocytes maintains glycolysis-dependent chemotaxis and VEGFA expression, enabling CCR2+ monocyte recruitment to metastatic sites; host APBA3 induces VEGFA-mediated E-selectin expression in endothelial cells of target organs, promoting cancer cell extravasation and micrometastasis. E-selectin-neutralizing antibody abolished host APBA3-mediated metastatic formation.","method":"APBA3-deficient mice, bone marrow transplant, chemotaxis assay, VEGFA/E-selectin measurement, neutralizing antibody experiment, experimental metastasis model","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — KO mice, conditional bone marrow transplant, neutralizing antibody rescue, multiple orthogonal in vivo and in vitro methods","pmids":["28507122"],"is_preprint":false},{"year":2021,"finding":"The N-terminal region of Mint3 (approximately residues 78–88) is intrinsically disordered but undergoes a disorder-to-order transition upon binding FIH-1, with large enthalpy and entropy changes consistent with coupled folding-and-binding. Residues 78–88 constitute the core binding site for FIH-1, while flanking disordered regions contribute enthalpically without increasing affinity.","method":"Circular dichroism, NMR, hydrogen/deuterium exchange mass spectrometry, isothermal titration calorimetry with truncation mutants","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 / Moderate — multiple structural and biophysical methods (CD, NMR, HDX-MS, ITC with mutagenesis), single lab but orthogonal methods","pmids":["34655613"],"is_preprint":false},{"year":2025,"finding":"Mint3 interacts directly with STING, selectively enhances K63-linked polyubiquitination of STING, and facilitates STING translocation from the ER to the Golgi, which enhances STING–TBK1 interaction and downstream IRF3 activation and type I IFN production in response to HSV-1 infection and cytosolic DNA stimulation.","method":"Co-immunoprecipitation (MINT3–STING interaction), K63-polyubiquitination assay, STING translocation assay by microscopy, TBK1 interaction assay, Mint3 KO/knockdown in macrophages and in vivo viral challenge","journal":"Cellular signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus ubiquitination assay plus localization assay plus KO mice, single lab, multiple orthogonal methods but not yet replicated","pmids":["40254147"],"is_preprint":false},{"year":2020,"finding":"Mint3 promotes transcription of the oncogenic ubiquitin ligase SKP2 via HIF-1, and this Mint3–HIF-1–SKP2 axis accumulates cell cycle inhibitors p21 and p27 when depleted, supporting cancer cell proliferation, EMT, stemness, and chemoresistance in pancreatic cancer cells.","method":"shRNA knockdown, gene expression analysis, HIF-1 reporter, cell proliferation and chemoresistance assays, orthotopic xenograft mouse model","journal":"Oncogene","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA depletion with multiple downstream readouts and in vivo xenograft, single lab","pmids":["32826949"],"is_preprint":false},{"year":2023,"finding":"Mint3 depletion reduces glycolytic adaptation of TNBC tumors to the microenvironment, causing energy stress that inactivates HSF1 via the AMPK/mTOR pathway, leading to decreased HSP70 expression and sensitization of TNBC tumors to chemotherapy in vivo.","method":"shRNA depletion, transcriptome analysis, AMPK/mTOR pathway assay, HSP70 inhibitor treatment, in vivo tumor model","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — shRNA plus transcriptome plus pathway rescue plus in vivo model, single lab","pmids":["38081808"],"is_preprint":false}],"current_model":"APBA3/Mint3 is a multifunctional adaptor protein that: (1) binds APP via its PTB domain and regulates APP export from the trans-Golgi network to the basolateral/cell-surface route, suppressing amyloidogenic processing; (2) constitutively inhibits FIH-1 (factor inhibiting HIF-1) through direct binding of its intrinsically disordered N-terminal region (core site residues 78–88), thereby sustaining normoxic HIF-1 transcriptional activity and driving the Warburg-type glycolytic metabolism in macrophages, cancer cells, and fibroblasts; (3) promotes K63-linked polyubiquitination of TRAF3 and STING, enhancing IRF3/IFN-β antiviral responses; (4) regulates TGN localization of Furin and surface recycling of MT5-MMP via PDZ domain interactions; and (5) links Rab6A GTPase to APP vesicle trafficking at the Golgi."},"narrative":{"mechanistic_narrative":"APBA3/Mint3 (X11L2) is a multifunctional Golgi-associated adaptor protein that couples vesicular trafficking decisions to metabolic and immune signaling [PMID:19726677, PMID:17959829]. Through its PTB domain it binds the intracellular YENPTY motif of APP via Tyr-682, marking APP-containing vesicles for export from the trans-Golgi network along the basolateral/cell-surface route rather than the endosomal/lysosomal route, thereby suppressing amyloidogenic Aβ generation [PMID:10049767, PMID:17959829, PMID:23965993]; this Golgi trafficking activity is linked to GTP-loaded Rab6A, which binds the complete PTB domain and co-localizes with Mint3 and APP at Golgi membranes [PMID:16207088, PMID:20447381]. Its tandem PDZ domains direct other cargoes, retrieving internalized MT5-MMP to the plasma membrane via its C-terminal EWV motif and retaining Furin at the TGN [PMID:14990567, PMID:18544638]. In parallel, the intrinsically disordered N-terminal region of Mint3 (core residues 78–88) directly binds and constitutively inhibits FIH-1 in an oxygen-independent manner through a coupled folding-and-binding mechanism, blocking FIH-1-mediated hydroxylation of HIF-1α Asn803 and sustaining normoxic HIF-1 transcriptional activity to drive Warburg-type glycolytic ATP production [PMID:19726677, PMID:34655613]. Genetic deletion in mice establishes this FIH-1–HIF-1 axis as the basis of a cell-type-specific role in macrophage energy metabolism, with knockout animals showing reduced glycolysis, cytokine production, and resistance to LPS-induced septic shock [PMID:21778228]. This glycolytic program supports inflammatory monocyte chemotaxis and VEGFA-driven endothelial E-selectin expression that promotes cancer cell extravasation and metastasis [PMID:28507122], and in tumor cells the Mint3–HIF-1 axis sustains proliferation through SKP2 induction and protects against chemotherapy-induced energy stress [PMID:32826949, PMID:38081808]. Independently of metabolism, Mint3 promotes K63-linked polyubiquitination of TRAF3 and STING to enhance IRF3 activation and type I IFN antiviral responses [PMID:27698125, PMID:40254147].","teleology":[{"year":1999,"claim":"Established APBA3 as a direct binding partner of APP, defining its candidate role in APP biology before any trafficking function was known.","evidence":"GST pull-down and reciprocal Co-IP of the PTB domain with the APP intracellular domain","pmids":["10049767"],"confidence":"Medium","gaps":["Functional consequence of the interaction not addressed","Domain mapping to specific APP motifs not yet resolved"]},{"year":2004,"claim":"Showed the PDZ domains can engage cargo independent of the PTB/APP axis, expanding Mint3 to a multi-cargo adaptor that controls surface recycling.","evidence":"Yeast two-hybrid, EWV motif deletion, and siRNA knockdown measuring MT5-MMP surface activity; separate PDZ binding/co-localization for Bcr","pmids":["14990567","15494376"],"confidence":"High","gaps":["Bcr interaction is Low-confidence with no functional follow-up","How PDZ cargo selection is coordinated with PTB cargo unknown"]},{"year":2005,"claim":"Linked Mint3 to a small GTPase regulator of Golgi trafficking, providing a mechanism for how APP vesicles are organized at the Golgi.","evidence":"Yeast two-hybrid, GTP-dependency assay, confocal co-localization of Rab6A/Mint3/APP, later confirmed by live-cell FRET with PTB-domain mutants","pmids":["16207088","20447381"],"confidence":"Medium","gaps":["No reciprocal Co-IP in mammalian cells in original report","Directionality of Rab6A regulation of Mint3 versus the reverse not established"]},{"year":2007,"claim":"Defined Mint3 as a determinant of post-Golgi APP sorting fate, mechanistically tying the adaptor to amyloidogenic processing.","evidence":"APP-vesicle purification, siRNA knockdown rerouting APP, and Aβ(1-40) ELISA; parallel work mapped nucleo-cytoplasmic shuttling and a transcriptional activator activity","pmids":["17959829","18201694"],"confidence":"High","gaps":["Mechanism connecting the nuclear shuttling/transcriptional activity to cytoplasmic trafficking roles unclear","Endogenous transcriptional targets of nuclear Mint3 not identified"]},{"year":2008,"claim":"Generalized Mint3's PTB-domain adaptor function to TGN retention of Furin, showing it organizes the localization of multiple secretory-pathway proteins.","evidence":"Co-IP, immunofluorescence, siRNA knockdown with Furin redistribution, and domain mapping in HeLa cells","pmids":["18544638"],"confidence":"Medium","gaps":["Whether Furin and APP compete for the same PTB site unknown","Physiological consequence of Furin mislocalization not measured"]},{"year":2009,"claim":"Identified a metabolic function entirely separate from trafficking: constitutive, oxygen-independent inhibition of FIH-1 to sustain normoxic HIF-1 activity and glycolysis.","evidence":"Purified-protein FIH-1 inhibition assay, HIF-1 reporter, siRNA knockdown, and biochemical fractionation in macrophages","pmids":["19726677"],"confidence":"High","gaps":["Whether the same molecule performs trafficking and FIH-1 inhibition simultaneously unresolved","Structural basis of FIH-1 binding not yet defined at this stage"]},{"year":2011,"claim":"Validated the FIH-1–HIF-1 axis in vivo and assigned it a cell-type-specific role in macrophage energetics and inflammation.","evidence":"Germline and myeloid-specific Apba3 knockout mice with ATP, glycolysis, cytokine readouts and an LPS septic shock model","pmids":["21778228"],"confidence":"High","gaps":["Contribution of trafficking functions to the in vivo phenotype not dissected","Non-myeloid roles of the axis untested here"]},{"year":2013,"claim":"Pinpointed the APP motif and residue (Tyr-682) that recruits Mint3 to the Golgi for APP export, refining the trafficking mechanism.","evidence":"Systematic site-directed mutagenesis of APP sorting motifs and subcellular localization assays","pmids":["23965993"],"confidence":"Medium","gaps":["LAMP1+ destination characterization incomplete","Single-lab finding"]},{"year":2014,"claim":"Demonstrated cell-type dependence of the FIH-1–Mint3 axis, showing it does not govern endogenous HIF-1 activity in all tissues.","evidence":"HIF-1 reporter with Mint3 N-terminus domain mapping and microarray after FIH-1 silencing in nucleus pulposus cells","pmids":["24867948"],"confidence":"Medium","gaps":["Reason for cell-type specificity of the axis unexplained","Negative endogenous result based on transcript profiling only"]},{"year":2016,"claim":"Uncovered a ubiquitin-dependent immune signaling role, expanding Mint3 from metabolism into antiviral and inflammatory signaling.","evidence":"Mint3 KO/knockdown macrophages with K63-polyubiquitination assays on TRAF3, IRF3/IFN-β readouts, and NF-κB/AMPK pathway analysis in influenza models","pmids":["27698125","27883071"],"confidence":"Medium","gaps":["How Mint3 promotes K63 ubiquitination mechanistically (E3 partner) unknown","Relationship between immune signaling and metabolic functions of Mint3 unclear"]},{"year":2017,"claim":"Connected the macrophage glycolytic program to pathophysiology, showing host Mint3 drives metastatic niche formation.","evidence":"APBA3 KO mice, bone marrow transplant, chemotaxis assays, VEGFA/E-selectin measurement and E-selectin neutralizing-antibody rescue in metastasis models","pmids":["28507122"],"confidence":"High","gaps":["Whether tumor-cell-intrinsic Mint3 contributes alongside host Mint3 not fully separated here"]},{"year":2021,"claim":"Provided the structural mechanism for FIH-1 inhibition, defining a disorder-to-order coupled folding-and-binding core at residues 78–88.","evidence":"CD, NMR, HDX-MS and ITC with truncation mutants","pmids":["34655613"],"confidence":"High","gaps":["No co-crystal/complex structure of the bound state","How binding sterically blocks FIH-1 catalysis not directly visualized"]},{"year":2023,"claim":"Extended the Mint3–HIF-1 glycolytic axis to tumor-cell-intrinsic survival, linking it to AMPK/mTOR/HSF1 stress responses and chemoresistance.","evidence":"shRNA depletion, transcriptome and AMPK/mTOR/HSP70 pathway analysis in TNBC in vivo models; parallel SKP2 axis in pancreatic cancer","pmids":["38081808","32826949"],"confidence":"Medium","gaps":["Direct versus indirect effects on HSF1/SKP2 not fully separated","Single-lab tumor models"]},{"year":2025,"claim":"Added STING to the Mint3 ubiquitination/trafficking repertoire, unifying its trafficking and immune-signaling roles in cytosolic DNA sensing.","evidence":"Co-IP, K63-polyubiquitination assay, STING ER-to-Golgi translocation and TBK1 interaction assays, KO macrophages and in vivo HSV-1 challenge","pmids":["40254147"],"confidence":"Medium","gaps":["Not independently replicated","Whether Golgi-trafficking machinery (Rab6A) participates in STING relocation untested"]},{"year":null,"claim":"How a single adaptor partitions between its Golgi-trafficking, FIH-1/HIF-1 metabolic, and ubiquitin-dependent immune functions—and how these are coordinated within one cell—remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No integrated model linking PTB/PDZ trafficking to N-terminal FIH-1 inhibition","E3 ligase mediating Mint3-dependent K63 ubiquitination unidentified","Structure of Mint3 in complex with any partner unsolved"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,3,5,6]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[1,15]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[8]}],"localization":[{"term_id":"GO:0005794","term_label":"Golgi apparatus","supporting_discovery_ids":[4,5,6,9,16]},{"term_id":"GO:0031410","term_label":"cytoplasmic vesicle","supporting_discovery_ids":[2]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[1,8]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[8]}],"pathway":[{"term_id":"R-HSA-9609507","term_label":"Protein localization","supporting_discovery_ids":[2,3,5,6]},{"term_id":"R-HSA-5653656","term_label":"Vesicle-mediated transport","supporting_discovery_ids":[2,6,16]},{"term_id":"R-HSA-1430728","term_label":"Metabolism","supporting_discovery_ids":[1,7,18]},{"term_id":"R-HSA-168256","term_label":"Immune System","supporting_discovery_ids":[12,13,16]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[1,7]}],"complexes":[],"partners":["APP","FIH1","RAB6A","MMP24","FURIN","TRAF3","STING1","BCR"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"O96018","full_name":"Amyloid-beta A4 precursor protein-binding family A member 3","aliases":["Adapter protein X11gamma","Neuron-specific X11L2 protein","Neuronal Munc18-1-interacting protein 3","Mint-3"],"length_aa":575,"mass_kda":61.5,"function":"May modulate processing of the amyloid-beta precursor protein (APP) and hence formation of APP-beta. May enhance the activity of HIF1A in macrophages by inhibiting the activity of HIF1AN","subcellular_location":"Cytoplasm, perinuclear region","url":"https://www.uniprot.org/uniprotkb/O96018/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/APBA3","classification":"Not Classified","n_dependent_lines":0,"n_total_lines":1208,"dependency_fraction":0.0},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/APBA3","total_profiled":1310},"omim":[{"mim_id":"604262","title":"AMYLOID BETA A4 PRECURSOR PROTEIN-BINDING, FAMILY A, MEMBER 3; APBA3","url":"https://www.omim.org/entry/604262"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Vesicles","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in many","driving_tissues":[],"url":"https://www.proteinatlas.org/search/APBA3"},"hgnc":{"alias_symbol":["X11L2","mint3"],"prev_symbol":[]},"alphafold":{"accession":"O96018","domains":[{"cath_id":"2.30.29.30","chopping":"213-311_326-361","consensus_level":"high","plddt":86.8525,"start":213,"end":361},{"cath_id":"2.30.42.10","chopping":"389-479","consensus_level":"high","plddt":84.6825,"start":389,"end":479},{"cath_id":"2.30.42.10","chopping":"485-565","consensus_level":"high","plddt":86.0022,"start":485,"end":565}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/O96018","model_url":"https://alphafold.ebi.ac.uk/files/AF-O96018-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-O96018-F1-predicted_aligned_error_v6.png","plddt_mean":64.44},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=APBA3","jax_strain_url":"https://www.jax.org/strain/search?query=APBA3"},"sequence":{"accession":"O96018","fasta_url":"https://rest.uniprot.org/uniprotkb/O96018.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/O96018/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/O96018"}},"corpus_meta":[{"pmid":"10049767","id":"PMC_10049767","title":"X11L2, a new member of the X11 protein family, interacts with Alzheimer's beta-amyloid precursor protein.","date":"1999","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/10049767","citation_count":74,"is_preprint":false},{"pmid":"19726677","id":"PMC_19726677","title":"Mint3 enhances the activity of hypoxia-inducible factor-1 (HIF-1) in macrophages by suppressing the activity of factor inhibiting HIF-1.","date":"2009","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/19726677","citation_count":55,"is_preprint":false},{"pmid":"17959829","id":"PMC_17959829","title":"Mint3/X11gamma is an ADP-ribosylation factor-dependent adaptor that regulates the traffic of the Alzheimer's Precursor protein from the trans-Golgi network.","date":"2007","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/17959829","citation_count":49,"is_preprint":false},{"pmid":"14990567","id":"PMC_14990567","title":"Mint-3 regulates the retrieval of the internalized membrane-type matrix metalloproteinase, MT5-MMP, to the plasma membrane by binding to its carboxyl end motif EWV.","date":"2004","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/14990567","citation_count":46,"is_preprint":false},{"pmid":"16207088","id":"PMC_16207088","title":"Rab6 interacts with the mint3 adaptor protein.","date":"2005","source":"Biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/16207088","citation_count":35,"is_preprint":false},{"pmid":"23965993","id":"PMC_23965993","title":"Recruitment of the Mint3 adaptor is necessary for export of the amyloid precursor protein (APP) from the Golgi complex.","date":"2013","source":"The Journal of biological 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America","url":"https://pubmed.ncbi.nlm.nih.gov/28507122","citation_count":28,"is_preprint":false},{"pmid":"29274998","id":"PMC_29274998","title":"DNA methylation of APBA3 and MCF2 in borderline personality disorder: Potential biomarkers for response to psychotherapy.","date":"2017","source":"European neuropsychopharmacology : the journal of the European College of Neuropsychopharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/29274998","citation_count":27,"is_preprint":false},{"pmid":"27698125","id":"PMC_27698125","title":"Mint3 potentiates TLR3/4- and RIG-I-induced IFN-β expression and antiviral immune responses.","date":"2016","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/27698125","citation_count":25,"is_preprint":false},{"pmid":"28504692","id":"PMC_28504692","title":"Mint3-mediated L1CAM expression in fibroblasts promotes cancer cell proliferation via integrin α5β1 and tumour growth.","date":"2017","source":"Oncogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/28504692","citation_count":25,"is_preprint":false},{"pmid":"32826949","id":"PMC_32826949","title":"Mint3 depletion restricts tumor malignancy of pancreatic cancer cells by decreasing SKP2 expression via HIF-1.","date":"2020","source":"Oncogene","url":"https://pubmed.ncbi.nlm.nih.gov/32826949","citation_count":22,"is_preprint":false},{"pmid":"15494376","id":"PMC_15494376","title":"Bcr (breakpoint cluster region) protein binds to PDZ-domains of scaffold protein PDZK1 and vesicle coat protein Mint3.","date":"2004","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/15494376","citation_count":22,"is_preprint":false},{"pmid":"24867948","id":"PMC_24867948","title":"FIH-1-Mint3 axis does not control HIF-1 transcriptional activity in nucleus pulposus cells.","date":"2014","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/24867948","citation_count":19,"is_preprint":false},{"pmid":"27883071","id":"PMC_27883071","title":"Mint3/Apba3 depletion ameliorates severe murine influenza pneumonia and macrophage cytokine production in response to the influenza virus.","date":"2016","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/27883071","citation_count":17,"is_preprint":false},{"pmid":"10574372","id":"PMC_10574372","title":"Genomic organization of the human X11L2 gene (APBA3), a third member of the X11 protein family interacting with Alzheimer's beta-amyloid precursor protein.","date":"1999","source":"Neuroreport","url":"https://pubmed.ncbi.nlm.nih.gov/10574372","citation_count":15,"is_preprint":false},{"pmid":"18201694","id":"PMC_18201694","title":"The X11L/X11beta/MINT2 and X11L2/X11gamma/MINT3 scaffold proteins shuttle between the nucleus and cytoplasm.","date":"2007","source":"Experimental cell research","url":"https://pubmed.ncbi.nlm.nih.gov/18201694","citation_count":14,"is_preprint":false},{"pmid":"28634075","id":"PMC_28634075","title":"Mint3 in bone marrow-derived cells promotes lung metastasis in breast cancer model mice.","date":"2017","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/28634075","citation_count":10,"is_preprint":false},{"pmid":"36831085","id":"PMC_36831085","title":"Mint3 as a Potential Target for Cooling Down HIF-1α-Mediated Inflammation and Cancer Aggressiveness.","date":"2023","source":"Biomedicines","url":"https://pubmed.ncbi.nlm.nih.gov/36831085","citation_count":9,"is_preprint":false},{"pmid":"20447381","id":"PMC_20447381","title":"Characterizing the interaction between the Rab6 GTPase and Mint3 via flow cytometry based FRET analysis.","date":"2010","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/20447381","citation_count":9,"is_preprint":false},{"pmid":"38081808","id":"PMC_38081808","title":"Mint3-depletion-induced energy stress sensitizes triple-negative breast cancer to chemotherapy via HSF1 inactivation.","date":"2023","source":"Cell death & disease","url":"https://pubmed.ncbi.nlm.nih.gov/38081808","citation_count":8,"is_preprint":false},{"pmid":"34655613","id":"PMC_34655613","title":"Structural and thermodynamical insights into the binding and inhibition of FIH-1 by the N-terminal disordered region of Mint3.","date":"2021","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/34655613","citation_count":8,"is_preprint":false},{"pmid":"33114509","id":"PMC_33114509","title":"Association of an APBA3 Missense Variant with Risk of Premature Ovarian Failure in the Korean Female Population.","date":"2020","source":"Journal of personalized medicine","url":"https://pubmed.ncbi.nlm.nih.gov/33114509","citation_count":6,"is_preprint":false},{"pmid":"40254147","id":"PMC_40254147","title":"MINT3 promotes STING activation and facilitates antiviral immune responses.","date":"2025","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/40254147","citation_count":2,"is_preprint":false},{"pmid":"33319072","id":"PMC_33319072","title":"Mint3 is dispensable for pancreatic and kidney functions in mice.","date":"2020","source":"Biochemistry and biophysics reports","url":"https://pubmed.ncbi.nlm.nih.gov/33319072","citation_count":1,"is_preprint":false},{"pmid":"40003897","id":"PMC_40003897","title":"Mint3 as a Molecular Target Activated in the Early Stage of Hepatocarcinogenesis.","date":"2025","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40003897","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14517,"output_tokens":4804,"usd":0.057806,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":12992,"output_tokens":4740,"usd":0.09173,"stage2_stop_reason":"end_turn"},"total_usd":0.149536,"stage1_batch_id":"msgbatch_0179Z2pxRYBsXi2xADneafRw","stage2_batch_id":"msgbatch_01LaY9ZaVdgjWArcZo9eqQE5","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1999,\n      \"finding\": \"APBA3/X11L2 PID/PTB domain directly interacts with the intracellular domain of APP (amyloid precursor protein), as demonstrated by GST binding assay in vitro and confirmed by co-immunoprecipitation from cells overexpressing APP and HA-tagged X11L2.\",\n      \"method\": \"GST pull-down assay, co-immunoprecipitation\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP plus in vitro GST pull-down, single lab, two orthogonal methods\",\n      \"pmids\": [\"10049767\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Mint3/APBA3 binds directly to FIH-1 (factor inhibiting HIF-1) and inhibits FIH-1's ability to hydroxylate HIF-1α Asn803, thereby preventing FIH-1-mediated suppression of HIF-1 transcriptional activity in an oxygen-independent manner. In macrophages, this mechanism sustains HIF-1 activity during normoxia and supports glycolytic ATP production. Knockdown of Mint3 redistributes FIH-1 from the perinuclear region to the cytoplasm and decreases glycolysis and ATP production.\",\n      \"method\": \"Purified protein binding assay (in vitro), HIF-1 reporter assay, siRNA knockdown, immunofluorescence co-localization, biochemical fractionation\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro reconstituted inhibition assay plus reporter assay plus KD phenotype in macrophages, multiple orthogonal methods in one study\",\n      \"pmids\": [\"19726677\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Mint3 is specifically enriched in APP-containing vesicles purified from neuroblastoma cells, and depletion of Mint3 by siRNA redirects APP export from the basolateral/trans-Golgi route to the endosomal/lysosomal route; Mint3 overexpression decreases and siRNA knockdown increases Aβ(1-40) secretion, identifying Mint3 as a critical determinant of post-Golgi APP trafficking.\",\n      \"method\": \"Vesicle purification from cells, siRNA knockdown, subcellular localization/trafficking assay, Aβ ELISA\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — vesicle purification plus siRNA KD plus functional Aβ measurement, multiple orthogonal methods\",\n      \"pmids\": [\"17959829\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The PDZ domains of Mint-3 bind the carboxyl EWV motif of MT5-MMP (identified by yeast two-hybrid screening), and this interaction mediates retrieval of internalized MT5-MMP to the plasma membrane; deletion of EWV impairs recycling without affecting internalization, and siRNA-mediated knockdown of Mint-3 decreases MT5-MMP surface activity.\",\n      \"method\": \"Yeast two-hybrid, siRNA knockdown, cell surface activity assay, deletion mutagenesis\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — yeast two-hybrid interaction mapped to specific motif, mutagenesis of EWV, siRNA functional validation, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"14990567\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Rab6A GTPase interacts with Mint3 in a GTP-dependent manner, requiring the complete PTB domain of Mint3. Rab6A, Mint3, and APP co-localize at Golgi membranes in HeLa cells, suggesting Mint3 links Rab6A to APP trafficking.\",\n      \"method\": \"Yeast two-hybrid, confocal microscopy co-localization, density gradient centrifugation, GTP-dependency assay\",\n      \"journal\": \"Biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — yeast two-hybrid plus co-localization plus fractionation, single lab, no reciprocal Co-IP in mammalian cells\",\n      \"pmids\": [\"16207088\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Mint3 PTB domain interacts with the acidic peptide signal of Furin in HeLa cells (shown by co-immunoprecipitation and immunofluorescence), and Mint3 knockdown by RNAi disrupts TGN-specific localization of Furin, redistributing it to endosomes.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, siRNA knockdown, domain mapping/mutagenesis\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal Co-IP plus siRNA KD with clear localization phenotype plus mutagenesis, single lab\",\n      \"pmids\": [\"18544638\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"Mint3 is recruited to the Golgi by the YENPTY motif of APP specifically via Tyr-682, and this recruitment is required for APP export from the Golgi; after leaving the Golgi, APP is directed to LAMP1+ structures as the proximal destination.\",\n      \"method\": \"Site-directed mutagenesis of APP sorting motifs, subcellular localization assay, adaptor recruitment assay\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — systematic mutagenesis of all four APP sorting motifs, specific residue identified, single lab\",\n      \"pmids\": [\"23965993\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Genetic deletion of Apba3 in mice reduces ATP production in macrophages to ~60% of wild-type, decreases glycolysis, cytokine production, and motility, and confers resistance to LPS-induced septic shock; myeloid-specific deletion recapitulates the septic shock resistance, establishing a cell-type-specific role for APBA3 in macrophage energy metabolism via the FIH-1–HIF-1 pathway.\",\n      \"method\": \"Apba3 knockout mice, myeloid-specific conditional knockout, ATP measurement, cytokine assay, LPS-induced septic shock model\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — germline and conditional KO mice with multiple functional readouts, mechanistic pathway confirmed in vivo\",\n      \"pmids\": [\"21778228\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"X11L2/APBA3 shuttles between the cytoplasm and nucleus via a nuclear export signal (NES) in its N-terminus; mutation of the NES causes nuclear accumulation, and X11L2 tethered near a promoter (via Gal4-DBD fusion) displays transcriptional activator activity, which is attenuated by preventing nucleo-cytoplasmic shuttling.\",\n      \"method\": \"Leptomycin B treatment, FLIP (fluorescence loss in photobleaching), NES mutagenesis, Gal4-DBD transcription reporter assay, nuclear fractionation\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — FLIP live imaging plus NES mutagenesis plus functional reporter assay, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"18201694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"The C-terminal PDZ-binding motif of Bcr (breakpoint cluster region protein) specifically binds to the PDZ domains of Mint3, localizing Bcr to the Golgi compartment where Mint3 resides.\",\n      \"method\": \"PDZ domain binding assay, co-localization immunofluorescence\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — PDZ binding assay and co-localization, single lab, single method type, limited mechanistic follow-up for Mint3 specifically\",\n      \"pmids\": [\"15494376\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"The PTB domain of Mint3 (Mint3Δ6) is sufficient to interact with constitutively active (GTP-loaded) Rab6A in living mammalian cells, as confirmed by FACS-based FRET analysis; a mutant lacking part of the PTB domain (Mint3Δ4) fails to interact.\",\n      \"method\": \"FACS-based FRET analysis in live cells, GST pull-down\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — FRET in live cells plus GST pull-down with domain-mapping mutants, single lab, two orthogonal methods\",\n      \"pmids\": [\"20447381\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In nucleus pulposus (NP) cells, overexpressed FIH-1 suppresses HIF-1 activity, and co-transfection of full-length Mint3 or the Mint3 N-terminus abrogates this suppression under both normoxia and hypoxia. However, endogenous FIH-1 silencing does not significantly change classical HIF-1 target gene transcripts in NP cells, indicating the FIH-1–Mint3 axis does not control endogenous HIF-1 transcriptional activity in this cell type.\",\n      \"method\": \"HIF-1 reporter assay, co-transfection, nuclear import/export inhibitors, microarray after FIH-1 silencing\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reporter assay plus domain mapping plus microarray, single lab; notable that this is a negative result for NP cell physiology\",\n      \"pmids\": [\"24867948\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Mint3 promotes K63-linked polyubiquitination of TRAF3, thereby enhancing IRF3 activation and IFN-β production downstream of TLR3/4 and RIG-I signaling in macrophages; Mint3 deficiency greatly attenuates antiviral immune responses and increases viral replication.\",\n      \"method\": \"Mint3 KO/knockdown in macrophages, ubiquitination assay (K63-linked), IRF3 activation assay, IFN-β reporter/ELISA, viral replication assay\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mice and KD with mechanistic ubiquitination assay, single lab\",\n      \"pmids\": [\"27698125\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In macrophages, Mint3 depletion attenuates NF-κB signaling (by increasing IκBα) and activates AMPK, reducing cytokine/chemokine production in response to influenza virus; Mint3-deficient mice show reduced inflammatory cytokines and neutrophil infiltration during influenza pneumonia without altering viral burden.\",\n      \"method\": \"Mint3 KO mice, siRNA knockdown, NF-κB pathway analysis (IκBα levels), AMPK activation assay, cytokine ELISA, neutrophil infiltration histology\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — KO mouse model plus in vitro pathway analysis with two distinct signaling readouts, single lab\",\n      \"pmids\": [\"27883071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Mint3/APBA3 in inflammatory monocytes maintains glycolysis-dependent chemotaxis and VEGFA expression, enabling CCR2+ monocyte recruitment to metastatic sites; host APBA3 induces VEGFA-mediated E-selectin expression in endothelial cells of target organs, promoting cancer cell extravasation and micrometastasis. E-selectin-neutralizing antibody abolished host APBA3-mediated metastatic formation.\",\n      \"method\": \"APBA3-deficient mice, bone marrow transplant, chemotaxis assay, VEGFA/E-selectin measurement, neutralizing antibody experiment, experimental metastasis model\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — KO mice, conditional bone marrow transplant, neutralizing antibody rescue, multiple orthogonal in vivo and in vitro methods\",\n      \"pmids\": [\"28507122\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"The N-terminal region of Mint3 (approximately residues 78–88) is intrinsically disordered but undergoes a disorder-to-order transition upon binding FIH-1, with large enthalpy and entropy changes consistent with coupled folding-and-binding. Residues 78–88 constitute the core binding site for FIH-1, while flanking disordered regions contribute enthalpically without increasing affinity.\",\n      \"method\": \"Circular dichroism, NMR, hydrogen/deuterium exchange mass spectrometry, isothermal titration calorimetry with truncation mutants\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — multiple structural and biophysical methods (CD, NMR, HDX-MS, ITC with mutagenesis), single lab but orthogonal methods\",\n      \"pmids\": [\"34655613\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"Mint3 interacts directly with STING, selectively enhances K63-linked polyubiquitination of STING, and facilitates STING translocation from the ER to the Golgi, which enhances STING–TBK1 interaction and downstream IRF3 activation and type I IFN production in response to HSV-1 infection and cytosolic DNA stimulation.\",\n      \"method\": \"Co-immunoprecipitation (MINT3–STING interaction), K63-polyubiquitination assay, STING translocation assay by microscopy, TBK1 interaction assay, Mint3 KO/knockdown in macrophages and in vivo viral challenge\",\n      \"journal\": \"Cellular signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus ubiquitination assay plus localization assay plus KO mice, single lab, multiple orthogonal methods but not yet replicated\",\n      \"pmids\": [\"40254147\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Mint3 promotes transcription of the oncogenic ubiquitin ligase SKP2 via HIF-1, and this Mint3–HIF-1–SKP2 axis accumulates cell cycle inhibitors p21 and p27 when depleted, supporting cancer cell proliferation, EMT, stemness, and chemoresistance in pancreatic cancer cells.\",\n      \"method\": \"shRNA knockdown, gene expression analysis, HIF-1 reporter, cell proliferation and chemoresistance assays, orthotopic xenograft mouse model\",\n      \"journal\": \"Oncogene\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA depletion with multiple downstream readouts and in vivo xenograft, single lab\",\n      \"pmids\": [\"32826949\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Mint3 depletion reduces glycolytic adaptation of TNBC tumors to the microenvironment, causing energy stress that inactivates HSF1 via the AMPK/mTOR pathway, leading to decreased HSP70 expression and sensitization of TNBC tumors to chemotherapy in vivo.\",\n      \"method\": \"shRNA depletion, transcriptome analysis, AMPK/mTOR pathway assay, HSP70 inhibitor treatment, in vivo tumor model\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — shRNA plus transcriptome plus pathway rescue plus in vivo model, single lab\",\n      \"pmids\": [\"38081808\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"APBA3/Mint3 is a multifunctional adaptor protein that: (1) binds APP via its PTB domain and regulates APP export from the trans-Golgi network to the basolateral/cell-surface route, suppressing amyloidogenic processing; (2) constitutively inhibits FIH-1 (factor inhibiting HIF-1) through direct binding of its intrinsically disordered N-terminal region (core site residues 78–88), thereby sustaining normoxic HIF-1 transcriptional activity and driving the Warburg-type glycolytic metabolism in macrophages, cancer cells, and fibroblasts; (3) promotes K63-linked polyubiquitination of TRAF3 and STING, enhancing IRF3/IFN-β antiviral responses; (4) regulates TGN localization of Furin and surface recycling of MT5-MMP via PDZ domain interactions; and (5) links Rab6A GTPase to APP vesicle trafficking at the Golgi.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"APBA3/Mint3 (X11L2) is a multifunctional Golgi-associated adaptor protein that couples vesicular trafficking decisions to metabolic and immune signaling [#1, #2]. Through its PTB domain it binds the intracellular YENPTY motif of APP via Tyr-682, marking APP-containing vesicles for export from the trans-Golgi network along the basolateral/cell-surface route rather than the endosomal/lysosomal route, thereby suppressing amyloidogenic Aβ generation [#0, #2, #6]; this Golgi trafficking activity is linked to GTP-loaded Rab6A, which binds the complete PTB domain and co-localizes with Mint3 and APP at Golgi membranes [#4, #10]. Its tandem PDZ domains direct other cargoes, retrieving internalized MT5-MMP to the plasma membrane via its C-terminal EWV motif and retaining Furin at the TGN [#3, #5]. In parallel, the intrinsically disordered N-terminal region of Mint3 (core residues 78–88) directly binds and constitutively inhibits FIH-1 in an oxygen-independent manner through a coupled folding-and-binding mechanism, blocking FIH-1-mediated hydroxylation of HIF-1α Asn803 and sustaining normoxic HIF-1 transcriptional activity to drive Warburg-type glycolytic ATP production [#1, #15]. Genetic deletion in mice establishes this FIH-1–HIF-1 axis as the basis of a cell-type-specific role in macrophage energy metabolism, with knockout animals showing reduced glycolysis, cytokine production, and resistance to LPS-induced septic shock [#7]. This glycolytic program supports inflammatory monocyte chemotaxis and VEGFA-driven endothelial E-selectin expression that promotes cancer cell extravasation and metastasis [#14], and in tumor cells the Mint3–HIF-1 axis sustains proliferation through SKP2 induction and protects against chemotherapy-induced energy stress [#17, #18]. Independently of metabolism, Mint3 promotes K63-linked polyubiquitination of TRAF3 and STING to enhance IRF3 activation and type I IFN antiviral responses [#12, #16].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Established APBA3 as a direct binding partner of APP, defining its candidate role in APP biology before any trafficking function was known.\",\n      \"evidence\": \"GST pull-down and reciprocal Co-IP of the PTB domain with the APP intracellular domain\",\n      \"pmids\": [\"10049767\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional consequence of the interaction not addressed\", \"Domain mapping to specific APP motifs not yet resolved\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Showed the PDZ domains can engage cargo independent of the PTB/APP axis, expanding Mint3 to a multi-cargo adaptor that controls surface recycling.\",\n      \"evidence\": \"Yeast two-hybrid, EWV motif deletion, and siRNA knockdown measuring MT5-MMP surface activity; separate PDZ binding/co-localization for Bcr\",\n      \"pmids\": [\"14990567\", \"15494376\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Bcr interaction is Low-confidence with no functional follow-up\", \"How PDZ cargo selection is coordinated with PTB cargo unknown\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Linked Mint3 to a small GTPase regulator of Golgi trafficking, providing a mechanism for how APP vesicles are organized at the Golgi.\",\n      \"evidence\": \"Yeast two-hybrid, GTP-dependency assay, confocal co-localization of Rab6A/Mint3/APP, later confirmed by live-cell FRET with PTB-domain mutants\",\n      \"pmids\": [\"16207088\", \"20447381\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No reciprocal Co-IP in mammalian cells in original report\", \"Directionality of Rab6A regulation of Mint3 versus the reverse not established\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Defined Mint3 as a determinant of post-Golgi APP sorting fate, mechanistically tying the adaptor to amyloidogenic processing.\",\n      \"evidence\": \"APP-vesicle purification, siRNA knockdown rerouting APP, and Aβ(1-40) ELISA; parallel work mapped nucleo-cytoplasmic shuttling and a transcriptional activator activity\",\n      \"pmids\": [\"17959829\", \"18201694\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism connecting the nuclear shuttling/transcriptional activity to cytoplasmic trafficking roles unclear\", \"Endogenous transcriptional targets of nuclear Mint3 not identified\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Generalized Mint3's PTB-domain adaptor function to TGN retention of Furin, showing it organizes the localization of multiple secretory-pathway proteins.\",\n      \"evidence\": \"Co-IP, immunofluorescence, siRNA knockdown with Furin redistribution, and domain mapping in HeLa cells\",\n      \"pmids\": [\"18544638\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether Furin and APP compete for the same PTB site unknown\", \"Physiological consequence of Furin mislocalization not measured\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Identified a metabolic function entirely separate from trafficking: constitutive, oxygen-independent inhibition of FIH-1 to sustain normoxic HIF-1 activity and glycolysis.\",\n      \"evidence\": \"Purified-protein FIH-1 inhibition assay, HIF-1 reporter, siRNA knockdown, and biochemical fractionation in macrophages\",\n      \"pmids\": [\"19726677\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the same molecule performs trafficking and FIH-1 inhibition simultaneously unresolved\", \"Structural basis of FIH-1 binding not yet defined at this stage\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Validated the FIH-1–HIF-1 axis in vivo and assigned it a cell-type-specific role in macrophage energetics and inflammation.\",\n      \"evidence\": \"Germline and myeloid-specific Apba3 knockout mice with ATP, glycolysis, cytokine readouts and an LPS septic shock model\",\n      \"pmids\": [\"21778228\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Contribution of trafficking functions to the in vivo phenotype not dissected\", \"Non-myeloid roles of the axis untested here\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Pinpointed the APP motif and residue (Tyr-682) that recruits Mint3 to the Golgi for APP export, refining the trafficking mechanism.\",\n      \"evidence\": \"Systematic site-directed mutagenesis of APP sorting motifs and subcellular localization assays\",\n      \"pmids\": [\"23965993\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"LAMP1+ destination characterization incomplete\", \"Single-lab finding\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Demonstrated cell-type dependence of the FIH-1–Mint3 axis, showing it does not govern endogenous HIF-1 activity in all tissues.\",\n      \"evidence\": \"HIF-1 reporter with Mint3 N-terminus domain mapping and microarray after FIH-1 silencing in nucleus pulposus cells\",\n      \"pmids\": [\"24867948\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reason for cell-type specificity of the axis unexplained\", \"Negative endogenous result based on transcript profiling only\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Uncovered a ubiquitin-dependent immune signaling role, expanding Mint3 from metabolism into antiviral and inflammatory signaling.\",\n      \"evidence\": \"Mint3 KO/knockdown macrophages with K63-polyubiquitination assays on TRAF3, IRF3/IFN-β readouts, and NF-κB/AMPK pathway analysis in influenza models\",\n      \"pmids\": [\"27698125\", \"27883071\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How Mint3 promotes K63 ubiquitination mechanistically (E3 partner) unknown\", \"Relationship between immune signaling and metabolic functions of Mint3 unclear\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Connected the macrophage glycolytic program to pathophysiology, showing host Mint3 drives metastatic niche formation.\",\n      \"evidence\": \"APBA3 KO mice, bone marrow transplant, chemotaxis assays, VEGFA/E-selectin measurement and E-selectin neutralizing-antibody rescue in metastasis models\",\n      \"pmids\": [\"28507122\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether tumor-cell-intrinsic Mint3 contributes alongside host Mint3 not fully separated here\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Provided the structural mechanism for FIH-1 inhibition, defining a disorder-to-order coupled folding-and-binding core at residues 78–88.\",\n      \"evidence\": \"CD, NMR, HDX-MS and ITC with truncation mutants\",\n      \"pmids\": [\"34655613\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No co-crystal/complex structure of the bound state\", \"How binding sterically blocks FIH-1 catalysis not directly visualized\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Extended the Mint3–HIF-1 glycolytic axis to tumor-cell-intrinsic survival, linking it to AMPK/mTOR/HSF1 stress responses and chemoresistance.\",\n      \"evidence\": \"shRNA depletion, transcriptome and AMPK/mTOR/HSP70 pathway analysis in TNBC in vivo models; parallel SKP2 axis in pancreatic cancer\",\n      \"pmids\": [\"38081808\", \"32826949\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct versus indirect effects on HSF1/SKP2 not fully separated\", \"Single-lab tumor models\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Added STING to the Mint3 ubiquitination/trafficking repertoire, unifying its trafficking and immune-signaling roles in cytosolic DNA sensing.\",\n      \"evidence\": \"Co-IP, K63-polyubiquitination assay, STING ER-to-Golgi translocation and TBK1 interaction assays, KO macrophages and in vivo HSV-1 challenge\",\n      \"pmids\": [\"40254147\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Not independently replicated\", \"Whether Golgi-trafficking machinery (Rab6A) participates in STING relocation untested\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How a single adaptor partitions between its Golgi-trafficking, FIH-1/HIF-1 metabolic, and ubiquitin-dependent immune functions—and how these are coordinated within one cell—remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No integrated model linking PTB/PDZ trafficking to N-terminal FIH-1 inhibition\", \"E3 ligase mediating Mint3-dependent K63 ubiquitination unidentified\", \"Structure of Mint3 in complex with any partner unsolved\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 3, 5, 6]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [1, 15]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005794\", \"supporting_discovery_ids\": [4, 5, 6, 9, 16]},\n      {\"term_id\": \"GO:0031410\", \"supporting_discovery_ids\": [2]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [1, 8]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-9609507\", \"supporting_discovery_ids\": [2, 3, 5, 6]},\n      {\"term_id\": \"R-HSA-5653656\", \"supporting_discovery_ids\": [2, 6, 16]},\n      {\"term_id\": \"R-HSA-1430728\", \"supporting_discovery_ids\": [1, 7, 18]},\n      {\"term_id\": \"R-HSA-168256\", \"supporting_discovery_ids\": [12, 13, 16]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [1, 7]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"APP\", \"FIH1\", \"RAB6A\", \"MMP24\", \"FURIN\", \"TRAF3\", \"STING1\", \"BCR\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":7,"faith_pct":85.71428571428571}}