{"gene":"NUSAP1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2017,"finding":"NUSAP1 localizes to dynamic spindle microtubules in a chromosome-centric pattern near overlapping microtubules during metaphase and anaphase. Mass spectrometry identified a cell cycle-regulated interaction between NUSAP1 and the RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex. The SAP domain of NUSAP1 is implicated in substrate recognition consistent with a co-factor role in sumoylation during chromosome segregation.","method":"Endogenous Co-IP, mass spectrometry, live cell imaging, siRNA knockdown with Taxol-sensitivity phenotype","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal endogenous Co-IP with MS validation, localization by imaging, functional phenotype (Taxol response) with RanBP2 depletion, single lab but multiple orthogonal methods","pmids":["28900032"],"is_preprint":false},{"year":2023,"finding":"NUSAP1 contains two consensus SUMOylation sites within its DNA-binding and microtubule-binding domains, respectively. Site-specific mutagenesis revealed that each site selectively controls NUSAP1 localization and generates distinct mitotic defects and daughter-cell fates. NUSAP1 forms proximity ligation products with SUMO2/3 in a RANBP2-dependent manner at key mitotic sites.","method":"Site-directed mutagenesis, proximity ligation assay, cell line expression of NuSAP1 mutants, mitotic phenotyping","journal":"Cells","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — mutagenesis of defined sites with functional phenotyping and proximity ligation validation, single lab, multiple orthogonal methods","pmids":["37947624"],"is_preprint":false},{"year":2020,"finding":"NUSAP1 stabilizes ATR protein by promoting its sumoylation via the SAP domain, antagonizing ubiquitin-dependent proteolysis of ATR. The SAP domain of NUSAP1 was necessary for this sumoylation and for chemoresistance to TMZ and DOX in glioblastoma cells.","method":"Co-IP, sumoylation assay, ubiquitination assay, SAP domain deletion/mutation, siRNA knockdown, cell viability assay","journal":"Signal transduction and targeted therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — domain-specific mechanistic dissection with sumoylation/ubiquitination assays and functional readout, single lab","pmids":["32317623"],"is_preprint":false},{"year":2023,"finding":"NUSAP1 interacts with ILF2 and DHX9 (RNA-binding proteins). The microtubule-binding and charged helical domains of NUSAP1 are required for these protein-protein interactions. Depletion of NUSAP1 abolished ILF2-dependent suppression of R-loop accumulation and DNA damage in response to camptothecin, placing NUSAP1 upstream of ILF2 in DNA damage/R-loop regulation.","method":"Affinity purification-mass spectrometry, Co-IP, confocal colocalization, domain deletion analysis, siRNA knockdown with R-loop and DNA damage assays","journal":"International journal of molecular sciences","confidence":"High","confidence_rationale":"Tier 2 / Moderate — AP-MS interactome plus reciprocal Co-IP verified by confocal colocalization, domain mapping, and functional epistasis with DNA damage readout; single lab but multiple orthogonal methods","pmids":["37047232"],"is_preprint":false},{"year":2014,"finding":"NUSAP1 depletion suppresses double-strand DNA break repair via homologous recombination and single-strand annealing pathways, and causes centrosome duplication defects. These phenotypes are mediated through BRCA1 protein levels: NUSAP1 depletion decreases BRCA1 protein and reduces BRCA1 recruitment to ionizing radiation-induced foci; BRCA1 overexpression from a plasmid rescues the defects caused by NUSAP1 depletion.","method":"siRNA knockdown, DNA repair assays (HR, SSA), centrosome counting, immunofluorescence for BRCA1 foci, plasmid rescue experiment","journal":"Cancer biology & therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis rescue experiment plus multiple phenotypic assays, single lab","pmids":["24521615"],"is_preprint":false},{"year":2023,"finding":"NUSAP1 binds to c-Myc and HIF-1α, forming a transcription regulatory complex that localizes to the LDHA promoter region and enhances LDHA expression. Lactate stabilizes NUSAP1 protein by inducing lysine lactylation (Kla) modification, inhibiting NUSAP1 protein degradation, creating a feedforward loop.","method":"ChIP-seq, Co-IP, RNA-seq, xenograft mouse model, single-cell RNA-seq, CHIP-seq verification of complex at LDHA promoter","journal":"Cancer letters","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP-seq localization of complex at promoter plus Co-IP for binding partners, single lab, multiple methods","pmids":["37354982"],"is_preprint":false},{"year":2021,"finding":"NUSAP1 functions as a positive regulator of YAP1 protein stability in gastric cancer cells. NUSAP1 was identified as a novel binding partner of YAP1 by Co-IP, and NUSAP1 depletion reduces YAP1 levels and downstream Hippo target gene transcription (CTGF, CYR61). Cancer-promoting effects of NUSAP1 on cell growth, migration and invasion are largely mediated through YAP1.","method":"Co-IP, siRNA knockdown, xenograft model, epistasis rescue (YAP1-dependent phenotype)","journal":"Frontiers in oncology","confidence":"Medium","confidence_rationale":"Tier 2-3 / Moderate — Co-IP identifying binding, plus functional epistasis showing YAP1 mediates NUSAP1 oncogenic effects, single lab","pmids":["33489890"],"is_preprint":false},{"year":2021,"finding":"NUSAP1 interacts with CDC20 and CCNA2 (Cyclin A2) by co-immunoprecipitation in osteosarcoma cells. NUSAP1 accelerates cell proliferation and cell cycle progression, and silencing either CDC20 or CCNA2 inhibits NUSAP1-induced proliferation and cell cycle progression.","method":"Co-immunoprecipitation, siRNA knockdown, CCK-8 and EdU proliferation assays, flow cytometry, xenograft model","journal":"OncoTargets and therapy","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — single Co-IP with functional epistasis by downstream knockdown, single lab","pmids":["34079289"],"is_preprint":false},{"year":2022,"finding":"NUSAP1 depletion in glioblastoma cells leads to downregulation of TOP2A, and silencing either NUSAP1 or TOP2A inhibits GBM cell proliferation and invasion and induces apoptosis. NUSAP1 expression is strongly correlated with TOP2A in glioma datasets, and stable NUSAP1 knockdown reduces tumor growth in an orthotopic xenograft model.","method":"RNA sequencing of NUSAP1-stable knockdown cells, siRNA knockdown of NUSAP1 and TOP2A, proliferation/invasion assays, orthotopic xenograft","journal":"Cell cycle (Georgetown, Tex.)","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — RNA-seq plus functional epistasis (double KD) in multiple cell lines and in vivo model, but no direct binding shown; single lab","pmids":["35532155"],"is_preprint":false},{"year":2015,"finding":"NUSAP1 expression is regulated by the RB1/E2F1 axis: RB1 knockdown increases NUSAP1 expression, and E2F1 knockdown decreases it. NUSAP1 knockdown in RB1-null or RB1-depleted prostate cancer cells decreases cell proliferation and invasion.","method":"Lentiviral RB1 knockdown, transient E2F1 siRNA, RT-qPCR, Western blot, proliferation and invasion assays","journal":"The Prostate","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis with two upstream factors (RB1, E2F1) and functional KD phenotype; single lab, multiple methods","pmids":["25585568"],"is_preprint":false},{"year":2022,"finding":"HOXB2 transcriptionally activates NUSAP1; dual-luciferase reporter and chromatin immunoprecipitation assays confirmed that HOXB2 binds the NUSAP1 promoter. HOXB2 overexpression abrogated the inhibitory effects of NUSAP1 silencing on nephroblastoma cell proliferation and metastasis via the PI3K/Akt signaling pathway.","method":"Dual-luciferase reporter assay, ChIP assay, JASPAR bioinformatics, Western blot for PI3K/Akt pathway proteins, siRNA and overexpression transfection","journal":"Molecular medicine reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP plus luciferase reporter verifying direct transcriptional regulation; functional epistasis with pathway readout; single lab","pmids":["35485274"],"is_preprint":false},{"year":2023,"finding":"Transcription factor E2F8 directly binds the NUSAP1 promoter (confirmed by ChIP and dual-luciferase reporter) and regulates NUSAP1 transcription. The E2F8/NUSAP1 axis promotes cisplatin resistance in hepatocellular carcinoma by reducing DNA damage (γ-H2AX), with NUSAP1 knockdown sensitizing cells to cisplatin.","method":"ChIP assay, dual-luciferase reporter, siRNA knockdown, CCK-8, flow cytometry, comet assay, Western blot","journal":"International journal of toxicology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP and luciferase reporter confirm direct transcriptional regulation; functional downstream assays; single lab","pmids":["37331996"],"is_preprint":false},{"year":2025,"finding":"PRMT1 catalyzes asymmetric dimethylation of NUSAP1 at R418 and R422 (ADMA). The R422 methylation site is critical for NUSAP1 function: methylated NUSAP1 interacts with the PEST domain of Notch2, inhibiting Notch2 ubiquitination and stabilizing Notch2 protein to activate Notch2 signaling. Inhibition of PRMT1 or R422 mutation abolishes NUSAP1-mediated Notch2 stabilization and 5-FU resistance in gastric cancer.","method":"Proteomic analysis, site-directed mutagenesis, Co-IP, ubiquitination assay, Western blot, functional resistance assays","journal":"Cell death & disease","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — mutagenesis of specific methylation sites with Co-IP and ubiquitination assay demonstrating mechanism; single lab, multiple methods","pmids":["40393971"],"is_preprint":false},{"year":2021,"finding":"O-GlcNAcylation (mediated by OGT) enhances NUSAP1 protein stability in bladder cancer cells. Pharmacological upregulation of O-GlcNAc increases NUSAP1 protein levels and stability, and OGT knockdown reduces tumor growth, while NUSAP1 overexpression impairs this effect, placing NUSAP1 downstream of OGT-mediated O-GlcNAcylation.","method":"Western blot, pharmacological O-GlcNAc modulation (GlcNAc, PuGNAc), OGT siRNA, CCK-8, flow cytometry, xenograft","journal":"OncoTargets and therapy","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, no direct demonstration of NUSAP1 O-GlcNAcylation site; stability inferred from protein levels after OGT manipulation","pmids":["33488099"],"is_preprint":false},{"year":2022,"finding":"ANKRD22 regulates NUSAP1 expression, and NUSAP1 overexpression reverses the inhibitory effects of ANKRD22 knockdown on breast cancer cell proliferation, invasion, and EMT. NUSAP1 mediates ANKRD22 activation of the Wnt/β-catenin signaling pathway.","method":"siRNA knockdown, overexpression rescue assay, Western blot for β-catenin pathway, BrdU, colony formation, Transwell assays","journal":"Bosnian journal of basic medical sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — epistasis rescue experiment placing NUSAP1 downstream of ANKRD22, single lab, no direct binding shown","pmids":["32651974"],"is_preprint":false},{"year":2022,"finding":"NUSAP1 promotes early recurrence of hepatocellular carcinoma by stimulating STAT3 nuclear translocation and activation through RACK1. RNA sequencing showed enrichment of cancer stemness and STAT3 pathway gene sets upon NUSAP1 overexpression.","method":"RNA sequencing, gain/loss-of-function experiments, in vivo recurrence mouse model, pathway analysis","journal":"Cancer science","confidence":"Low","confidence_rationale":"Tier 3 / Weak — STAT3 pathway activation proposed via RACK1 linkage from RNA-seq; mechanistic detail sparse in abstract; single lab","pmids":["36106345"],"is_preprint":false},{"year":2025,"finding":"NUSAP1 functions as a tumor suppressor in triple-negative breast cancer by recruiting the transcriptional repressor DAXX via its microtubule-associated domain (MAD) to bridge HIFα. This complex recruits SETDB1 methyltransferase and deposits H3K9me3 repressive marks on hypoxia response elements (HREs), attenuating HIF transcriptional activity and expression of HIF target genes.","method":"NUSAP1 depletion/overexpression, in vitro and in vivo invasion/proliferation assays, Co-IP (NUSAP1-DAXX-HIFα), ChIP for H3K9me3 at HREs, engineered MAD domain rescue","journal":"Advanced science (Weinheim, Baden-Wurttemberg, Germany)","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating ternary complex plus ChIP showing H3K9me3 deposition at HREs; functional domain-engineering rescue; single lab, multiple orthogonal methods","pmids":["41178464"],"is_preprint":false},{"year":2024,"finding":"NUSAP1 interacts with ANXA2 (Annexin A2) and protects it from ubiquitin-dependent protein degradation, thereby increasing ANXA2 stability. NUSAP1 silencing increases GC radiosensitivity by impairing DNA damage repair after irradiation.","method":"Immunoprecipitation, mass spectrometry, ubiquitination assay, colony formation, flow cytometry, comet assay, xenograft","journal":"Journal of cancer research and clinical oncology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — IP-MS identifies binding partner, ubiquitination assay demonstrates mechanism of stabilization, functional radiosensitivity readout; single lab","pmids":["39212774"],"is_preprint":false},{"year":2025,"finding":"NUSAP1 interacts with SHCBP1 and inhibits JAK2/STAT3 phosphorylation by blocking SHCBP1 in hepatocellular carcinoma cells, promoting dendritic cell generation from co-cultured PBMCs.","method":"Co-immunoprecipitation, Western blot for phospho-JAK2/STAT3, PBMC co-culture with HCC cells, flow cytometry for CD1a/CD86","journal":"Journal of immunotherapy","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single Co-IP with pathway readout; mechanistic detail limited in abstract; single lab","pmids":["38980111"],"is_preprint":false},{"year":2025,"finding":"NUSAP1 directly interacts with KIF2C (a kinesin-13 microtubule depolymerase) and Aurora kinase A (AURKA). AURKA-mediated phosphorylation of NUSAP1 modulates NUSAP1's binding affinity to KIF2C, balancing spindle microtubule stability and depolymerization. Truncating NUSAP1 mutations (Y403* and Q405*) impair NUSAP1-KIF2C interaction. NUSAP1 rescues cell cycle arrest and apoptosis caused by KIF2C overexpression in NUSAP1-KO cells, but mutant truncated NUSAP1 fails this rescue.","method":"Co-IP (NUSAP1-KIF2C, NUSAP1-AURKA), phosphorylation assay, NUSAP1-KO HEK293T rescue experiment, mouse microcephaly model, live cell imaging","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating direct interactions, phosphorylation assay, domain-disrupting mutation rescue experiment; preprint, single lab, multiple methods","pmids":["bio_10.1101_2025.05.16.654427"],"is_preprint":true},{"year":2023,"finding":"A recurrent de novo heterozygous nonsense variant (p.Tyr403Ter) in NUSAP1 causes microcephaly, severe developmental delay, and epilepsy. The mutant transcript escapes nonsense-mediated decay, suggesting dominant-negative or gain-of-function mechanism rather than haploinsufficiency.","method":"Exome sequencing, NMD escape assay (transcript analysis), single-cell RNA-seq of post-mortem brain","journal":"Clinical genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — NMD escape experimentally demonstrated; mechanism (dominant-negative vs. GOF) not yet resolved; single lab","pmids":["37005340"],"is_preprint":false},{"year":2019,"finding":"In zebrafish, nusap1 morpholino knockdown impairs neural crest cell migration, alters expression of neural crest markers crestin and sox9b, and causes apoptosis in retina and hindbrain. These phenotypes are rescued by co-injection of nusap1 mRNA. This establishes an in vivo role for Nusap1 in cell migration and neural crest morphogenesis.","method":"Antisense morpholino knockdown in zebrafish, in situ hybridization for neural crest markers, mRNA rescue injection","journal":"Protein & cell","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — morpholino KD with mRNA rescue (gold standard in zebrafish), multiple marker readouts; single lab","pmids":["21203972"],"is_preprint":false},{"year":2023,"finding":"In mouse oocytes, NUSAP1 localizes to distinct granular aggregates near spindle poles during Pro-MI, metaphase I, and anaphase I. NUSAP1 depletion causes chromosome misalignment, increased aneuploidy, abnormal spindle assembly with decreased spindle pole width, and attenuation of cortical F-actin. Mass spectrometry of NUSAP1 interactomes showed significant enrichment for RNA-binding partners, suggesting NUSAP1 influences P-body dynamics and mRNA homeostasis.","method":"Protein expression profiling (proteomics), siRNA depletion, immunofluorescence for localization, aneuploidy measurement, RNA-seq, mass spectrometry interactome","journal":"Journal of cellular biochemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct localization by imaging with functional depletion phenotype and MS interactome; single lab, multiple methods","pmids":["37992207"],"is_preprint":false},{"year":2025,"finding":"NUSAP1 interacts with NUSAP1-IP3R (inositol 1,4,5-trisphosphate receptor); NUSAP1 governs IP3R phosphorylation status to regulate cytosolic Ca2+ homeostasis and ER stress. NUSAP1 knockdown increases cytosolic Ca2+, p-IP3R, and ER stress, promoting mitophagy and apoptosis in endometrial carcinoma cells. IP3R silencing reduces Ca2+ and ER stress without affecting NUSAP1 expression, but NUSAP1 overexpression combined with IP3R knockdown amplified these effects synergistically.","method":"Co-IP (NUSAP1-IP3R interaction), Fluo-4 AM Ca2+ measurement, ER fluorescence probe imaging, flow cytometry, Western blot, siRNA knockdown and overexpression","journal":"ACS omega","confidence":"Low","confidence_rationale":"Tier 3 / Weak — Co-IP demonstrates interaction; IP3R phosphorylation mechanism inferred from pharmacological and genetic epistasis; single lab, no in vitro kinase/phosphatase assay","pmids":["41585655"],"is_preprint":false},{"year":2025,"finding":"HBV core protein (HBC) enhances WDR46 protein stability by blocking TRIM25-mediated ubiquitination of WDR46. WDR46 then facilitates c-Myc recruitment to the NUSAP1 promoter by enhancing the WDR46-c-Myc interaction, thereby transcriptionally upregulating NUSAP1 to promote HCC cell growth and migration.","method":"Co-immunoprecipitation, ChIP, Western blot, ubiquitination assay, functional growth and migration assays, xenograft model","journal":"Hepatology communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — ChIP confirms c-Myc recruitment to NUSAP1 promoter; Co-IP and ubiquitination assay delineate upstream WDR46 stabilization mechanism; single lab, multiple methods","pmids":["40366140"],"is_preprint":false},{"year":2024,"finding":"NUSAP1 promotes lung adenocarcinoma cell proliferation, and its upregulation is mediated by estrogen via ERβ activation. Pharmacological targeting of the ERβ/NUSAP1 axis with fulvestrant (ERβ antagonist) or entinostat (identified as a novel NUSAP1 inhibitor) suppresses LUAD growth in vitro and in vivo.","method":"Bioinformatics, in vitro proliferation assays with estrogen treatment, ERβ activation/inhibition, entinostat treatment, in vivo xenograft","journal":"International journal of biological sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — regulatory mechanism (estrogen-ERβ-NUSAP1) demonstrated pharmacologically; direct binding/transcriptional mechanism not fully delineated in abstract; single lab","pmids":["39430250"],"is_preprint":false}],"current_model":"NUSAP1 is a microtubule-associated protein that localizes to the mitotic spindle and plays key roles in spindle assembly, chromosome segregation, and DNA damage response: it interacts with the RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex via its SAP domain (which contains two functional SUMOylation consensus sites regulated by RANBP2), promotes ATR stability via SAP domain-mediated sumoylation, regulates BRCA1 protein levels to support DSB repair and centrosome duplication, suppresses R-loop accumulation through interaction with RNA-binding protein ILF2, and is phosphorylated by AURKA to modulate its interaction with KIF2C for spindle microtubule balance; in cancer contexts NUSAP1 forms a transcriptional complex with c-Myc and HIF-1α to drive LDHA expression, stabilizes YAP1 and ANXA2 proteins by suppressing their ubiquitination, and is itself transcriptionally controlled by the RB1/E2F1 axis and by HOXB2/E2F8; PRMT1-mediated dimethylation at R422 enables NUSAP1 to stabilize Notch2 via PEST domain interaction, while in triple-negative breast cancer NUSAP1 acts as a tumor suppressor by recruiting DAXX to HIFα and depositing H3K9me3 repressive marks at hypoxia response elements."},"narrative":{"mechanistic_narrative":"NUSAP1 is a microtubule-associated protein that localizes to dynamic spindle microtubules in a chromosome-centric pattern during metaphase and anaphase, where it organizes spindle architecture and chromosome segregation through SUMO-pathway and kinesin interactions [PMID:28900032, PMID:bio_10.1101_2025.05.16.654427]. Through its SAP domain it engages the RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex, and two consensus SUMOylation sites within its DNA- and microtubule-binding domains are modified in a RANBP2-dependent manner, each site selectively controlling NUSAP1 localization and dictating distinct mitotic outcomes [PMID:28900032, PMID:37947624]. NUSAP1 directly binds KIF2C and Aurora kinase A; AURKA-mediated phosphorylation tunes NUSAP1-KIF2C affinity to balance spindle microtubule stability against depolymerization, and truncating mutations (Y403*, Q405*) that disrupt this interaction abolish its rescue activity [PMID:bio_10.1101_2025.05.16.654427]. Beyond mitosis, NUSAP1 supports genome stability: it sustains BRCA1 protein levels and BRCA1 focus formation to enable homologous-recombination and single-strand-annealing repair and proper centrosome duplication [PMID:24521615], and it acts upstream of the RNA-binding protein ILF2 (also interacting with DHX9) to suppress R-loop accumulation and DNA damage [PMID:37047232]. Across cancers, NUSAP1 is deployed as an oncogenic protein-stabilizing and transcription-regulating hub—forming a c-Myc/HIF-1α complex at the LDHA promoter to drive glycolysis [PMID:37354982], stabilizing partner proteins including ATR, YAP1, ANXA2 and Notch2 by antagonizing their ubiquitin-dependent degradation [PMID:32317623, PMID:33489890, PMID:39212774, PMID:40393971], and is itself transcriptionally controlled by the RB1/E2F1 and E2F8 axes [PMID:25585568, PMID:37331996]. In triple-negative breast cancer, by contrast, NUSAP1 acts as a tumor suppressor, recruiting DAXX via its microtubule-associated domain to bridge HIFα and deposit SETDB1-dependent H3K9me3 marks on hypoxia response elements to repress HIF target genes [PMID:41178464]. A recurrent de novo nonsense variant (p.Tyr403Ter) that escapes nonsense-mediated decay causes microcephaly, severe developmental delay, and epilepsy [PMID:37005340], consistent with conserved developmental roles seen in zebrafish neural crest migration [PMID:21203972] and mouse oocyte spindle assembly [PMID:37992207].","teleology":[{"year":2014,"claim":"Established that NUSAP1 contributes to genome maintenance beyond its mitotic role by linking it to double-strand break repair and centrosome integrity.","evidence":"siRNA depletion with HR/SSA repair assays, BRCA1 focus imaging, and plasmid rescue in human cells","pmids":["24521615"],"confidence":"Medium","gaps":["Mechanism by which NUSAP1 controls BRCA1 protein level not defined","No direct NUSAP1-BRCA1 binding shown"]},{"year":2015,"claim":"Placed NUSAP1 transcription under the RB1/E2F1 axis, explaining its cell-cycle-coupled expression and elevation in RB1-deficient cancer.","evidence":"Lentiviral RB1 and siRNA E2F1 knockdown with RT-qPCR, Western blot, and proliferation/invasion assays in prostate cancer cells","pmids":["25585568"],"confidence":"Medium","gaps":["Direct E2F1 binding to the NUSAP1 promoter not demonstrated here","Does not address post-translational regulation"]},{"year":2017,"claim":"Identified the molecular basis of NUSAP1's mitotic function as a SUMO-pathway cofactor by mapping its interaction with the RanBP2-RanGAP1-UBC9 complex.","evidence":"Endogenous reciprocal Co-IP with mass spectrometry, live-cell imaging, and Taxol-sensitivity phenotyping in human cells","pmids":["28900032"],"confidence":"High","gaps":["Specific SUMO substrates recognized via the SAP domain not identified","Single lab"]},{"year":2019,"claim":"Demonstrated an in vivo developmental requirement for NUSAP1 in cell migration and neural crest morphogenesis.","evidence":"Zebrafish morpholino knockdown with neural crest marker in situ hybridization and mRNA rescue","pmids":["21203972"],"confidence":"Medium","gaps":["Molecular mechanism linking NUSAP1 to neural crest migration unresolved","Morpholino specificity limitations"]},{"year":2020,"claim":"Provided a substrate-level mechanism for SAP-domain function by showing NUSAP1 promotes ATR sumoylation to antagonize its proteolysis and confer chemoresistance.","evidence":"Co-IP, sumoylation and ubiquitination assays, SAP-domain mutation, and viability assays in glioblastoma cells","pmids":["32317623"],"confidence":"Medium","gaps":["Direct SUMO E3 activity vs cofactor role not separated","Single lab"]},{"year":2021,"claim":"Extended NUSAP1's protein-stabilizing role to cancer signaling hubs YAP1 and cell-cycle regulators CDC20/CCNA2 driving proliferation.","evidence":"Co-IP, siRNA knockdown, epistasis rescue, and xenografts in gastric cancer and osteosarcoma cells","pmids":["33489890","34079289"],"confidence":"Medium","gaps":["Whether NUSAP1 stabilizes YAP1 by blocking ubiquitination not directly tested","Direct binding surfaces undefined"]},{"year":2022,"claim":"Mapped additional transcriptional inputs (HOXB2) and downstream effectors (TOP2A) that integrate NUSAP1 into proliferative cancer programs.","evidence":"ChIP and dual-luciferase reporter for HOXB2, RNA-seq plus double knockdown for TOP2A, with xenograft models","pmids":["35485274","35532155"],"confidence":"Medium","gaps":["TOP2A regulation shown without direct binding","Tissue specificity of these axes unclear"]},{"year":2023,"claim":"Resolved the site-specific logic of NUSAP1 SUMOylation, showing two RANBP2-dependent sites independently govern localization and daughter-cell fate.","evidence":"Site-directed mutagenesis, proximity ligation with SUMO2/3, and mitotic phenotyping in human cells","pmids":["37947624"],"confidence":"High","gaps":["Downstream effectors of each modified site not identified"]},{"year":2023,"claim":"Connected NUSAP1 to RNA metabolism and R-loop suppression by placing it upstream of ILF2/DHX9 in the DNA damage response.","evidence":"AP-MS interactome, reciprocal Co-IP, confocal colocalization, domain mapping, and R-loop/DNA damage assays after camptothecin","pmids":["37047232"],"confidence":"High","gaps":["How NUSAP1 mechanistically enables ILF2 R-loop suppression unclear","Single lab"]},{"year":2023,"claim":"Defined a metabolic-transcriptional role in which NUSAP1 joins c-Myc and HIF-1α at the LDHA promoter, reinforced by lactylation-driven stabilization.","evidence":"ChIP-seq, Co-IP, RNA-seq, single-cell RNA-seq, and xenografts","pmids":["37354982"],"confidence":"Medium","gaps":["Lactylation site on NUSAP1 not mapped","Direct DNA contact by NUSAP1 vs partners unclear"]},{"year":2023,"claim":"Identified E2F8 as an additional direct transcriptional activator of NUSAP1 mediating cisplatin resistance.","evidence":"ChIP, dual-luciferase reporter, knockdown, comet assay, and γ-H2AX analysis in hepatocellular carcinoma","pmids":["37331996"],"confidence":"Medium","gaps":["Relationship to RB1/E2F1 axis not integrated"]},{"year":2023,"claim":"Confirmed conserved meiotic spindle and RNA-associated functions in mammalian oocytes, reinforcing the link between NUSAP1 and RNA-binding partners.","evidence":"Proteomics, siRNA depletion, immunofluorescence localization, aneuploidy scoring, and MS interactome in mouse oocytes","pmids":["37992207"],"confidence":"Medium","gaps":["Direct role in P-body dynamics inferred, not demonstrated","Specific RNA targets unidentified"]},{"year":2023,"claim":"Linked NUSAP1 to human neurodevelopmental disease through a recurrent de novo nonsense variant acting via a non-haploinsufficiency mechanism.","evidence":"Exome sequencing, NMD escape transcript assay, and single-cell RNA-seq of post-mortem brain","pmids":["37005340"],"confidence":"Medium","gaps":["Dominant-negative vs gain-of-function mechanism unresolved","Single family/lab cohort"]},{"year":2024,"claim":"Extended the protein-stabilization paradigm to ANXA2, tying NUSAP1 to radioresistance via DNA-damage repair.","evidence":"IP-MS, ubiquitination assay, comet assay, and xenografts in gastric cancer","pmids":["39212774"],"confidence":"Medium","gaps":["E3 ligase antagonized by NUSAP1 not identified"]},{"year":2025,"claim":"Established the direct AURKA-NUSAP1-KIF2C axis that balances spindle microtubule dynamics and tied disease-associated truncations to its disruption.","evidence":"Co-IP, phosphorylation assay, NUSAP1-KO rescue, mouse microcephaly model, and live imaging (preprint)","pmids":["bio_10.1101_2025.05.16.654427"],"confidence":"Medium","gaps":["Preprint, single lab","Phosphosites on NUSAP1 not fully mapped"]},{"year":2025,"claim":"Revealed a context-dependent tumor-suppressor function in which NUSAP1 represses HIF transcriptional output via DAXX/SETDB1-mediated H3K9me3 deposition.","evidence":"Co-IP of NUSAP1-DAXX-HIFα, ChIP for H3K9me3 at HREs, and MAD-domain rescue in triple-negative breast cancer","pmids":["41178464"],"confidence":"Medium","gaps":["Determinants of oncogenic vs tumor-suppressor switching unknown","Single lab"]},{"year":2025,"claim":"Added a PRMT1-dependent methylation layer (R418/R422) enabling NUSAP1 to stabilize Notch2 and drive 5-FU resistance.","evidence":"Proteomics, site-directed mutagenesis, Co-IP, and ubiquitination assays in gastric cancer","pmids":["40393971"],"confidence":"Medium","gaps":["Structural basis of methyl-dependent PEST-domain binding undefined"]},{"year":null,"claim":"What governs the switch between NUSAP1's oncogenic protein-stabilizing/transcriptional roles and its tumor-suppressive HIF-repressive role, and how its mitotic, DNA-repair, and disease-causing functions are mechanistically unified, remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying biochemical model across contexts","Structural basis of SAP/MAD domain partner selection unknown","Direct enzymatic vs adaptor role in SUMO and ubiquitin pathways not separated"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,19]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,16,19]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[5,16]},{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[2,17,12]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[1]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,22]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[5,16]},{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1,19,7]},{"term_id":"R-HSA-73894","term_label":"DNA 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MicroRNA-769-5p suppresses cell growth and migration via targeting NUSAP1 in bladder cancer. J Clin Lab Anal. 2020; 34:e23193.","date":"2023","source":"Journal of clinical laboratory analysis","url":"https://pubmed.ncbi.nlm.nih.gov/37183304","citation_count":1,"is_preprint":false},{"pmid":"40909470","id":"PMC_40909470","title":"NuSAP1 promotes bipolar spindle assembly in Trypanosoma brucei by bundling spindle microtubules.","date":"2025","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/40909470","citation_count":0,"is_preprint":false},{"pmid":"41908258","id":"PMC_41908258","title":"CCNB2, NUSAP1, GTSE1, and TK1: potential dual biomarkers for pneumonia and lung adenocarcinoma.","date":"2026","source":"Cytotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/41908258","citation_count":0,"is_preprint":false},{"pmid":"39639090","id":"PMC_39639090","title":"Multi-omics analysis detail a submicroscopic inv(15)(q14q15) generating fusion transcripts and MEIS2 and NUSAP1 haploinsufficiency.","date":"2024","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/39639090","citation_count":0,"is_preprint":false},{"pmid":"41275481","id":"PMC_41275481","title":"Research Article: In Silico Identification of Potential Inhibitors Targeting NUSAP1, A Hub Gene in Hepatocellular Carcinoma.","date":"2025","source":"Critical reviews in oncogenesis","url":"https://pubmed.ncbi.nlm.nih.gov/41275481","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.08.25.672125","title":"NuSAP1 promotes bipolar spindle assembly in  <i>Trypanosoma brucei</i>  by bundling spindle microtubules","date":"2025-08-25","source":"bioRxiv","url":"https://doi.org/10.1101/2025.08.25.672125","citation_count":0,"is_preprint":true},{"pmid":null,"id":"bio_10.1101_2025.05.16.654427","title":"NUSAP1 regulates mitotic processes via KIF2C interaction and AURKA phosphorylation in primary microcephaly","date":"2025-05-16","source":"bioRxiv","url":"https://doi.org/10.1101/2025.05.16.654427","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":36676,"output_tokens":7123,"usd":0.108437,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":16334,"output_tokens":5392,"usd":0.108235,"stage2_stop_reason":"end_turn"},"total_usd":0.216672,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2017,\n      \"finding\": \"NUSAP1 localizes to dynamic spindle microtubules in a chromosome-centric pattern near overlapping microtubules during metaphase and anaphase. Mass spectrometry identified a cell cycle-regulated interaction between NUSAP1 and the RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex. The SAP domain of NUSAP1 is implicated in substrate recognition consistent with a co-factor role in sumoylation during chromosome segregation.\",\n      \"method\": \"Endogenous Co-IP, mass spectrometry, live cell imaging, siRNA knockdown with Taxol-sensitivity phenotype\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal endogenous Co-IP with MS validation, localization by imaging, functional phenotype (Taxol response) with RanBP2 depletion, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"28900032\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NUSAP1 contains two consensus SUMOylation sites within its DNA-binding and microtubule-binding domains, respectively. Site-specific mutagenesis revealed that each site selectively controls NUSAP1 localization and generates distinct mitotic defects and daughter-cell fates. NUSAP1 forms proximity ligation products with SUMO2/3 in a RANBP2-dependent manner at key mitotic sites.\",\n      \"method\": \"Site-directed mutagenesis, proximity ligation assay, cell line expression of NuSAP1 mutants, mitotic phenotyping\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — mutagenesis of defined sites with functional phenotyping and proximity ligation validation, single lab, multiple orthogonal methods\",\n      \"pmids\": [\"37947624\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"NUSAP1 stabilizes ATR protein by promoting its sumoylation via the SAP domain, antagonizing ubiquitin-dependent proteolysis of ATR. The SAP domain of NUSAP1 was necessary for this sumoylation and for chemoresistance to TMZ and DOX in glioblastoma cells.\",\n      \"method\": \"Co-IP, sumoylation assay, ubiquitination assay, SAP domain deletion/mutation, siRNA knockdown, cell viability assay\",\n      \"journal\": \"Signal transduction and targeted therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — domain-specific mechanistic dissection with sumoylation/ubiquitination assays and functional readout, single lab\",\n      \"pmids\": [\"32317623\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NUSAP1 interacts with ILF2 and DHX9 (RNA-binding proteins). The microtubule-binding and charged helical domains of NUSAP1 are required for these protein-protein interactions. Depletion of NUSAP1 abolished ILF2-dependent suppression of R-loop accumulation and DNA damage in response to camptothecin, placing NUSAP1 upstream of ILF2 in DNA damage/R-loop regulation.\",\n      \"method\": \"Affinity purification-mass spectrometry, Co-IP, confocal colocalization, domain deletion analysis, siRNA knockdown with R-loop and DNA damage assays\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — AP-MS interactome plus reciprocal Co-IP verified by confocal colocalization, domain mapping, and functional epistasis with DNA damage readout; single lab but multiple orthogonal methods\",\n      \"pmids\": [\"37047232\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"NUSAP1 depletion suppresses double-strand DNA break repair via homologous recombination and single-strand annealing pathways, and causes centrosome duplication defects. These phenotypes are mediated through BRCA1 protein levels: NUSAP1 depletion decreases BRCA1 protein and reduces BRCA1 recruitment to ionizing radiation-induced foci; BRCA1 overexpression from a plasmid rescues the defects caused by NUSAP1 depletion.\",\n      \"method\": \"siRNA knockdown, DNA repair assays (HR, SSA), centrosome counting, immunofluorescence for BRCA1 foci, plasmid rescue experiment\",\n      \"journal\": \"Cancer biology & therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis rescue experiment plus multiple phenotypic assays, single lab\",\n      \"pmids\": [\"24521615\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NUSAP1 binds to c-Myc and HIF-1α, forming a transcription regulatory complex that localizes to the LDHA promoter region and enhances LDHA expression. Lactate stabilizes NUSAP1 protein by inducing lysine lactylation (Kla) modification, inhibiting NUSAP1 protein degradation, creating a feedforward loop.\",\n      \"method\": \"ChIP-seq, Co-IP, RNA-seq, xenograft mouse model, single-cell RNA-seq, CHIP-seq verification of complex at LDHA promoter\",\n      \"journal\": \"Cancer letters\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP-seq localization of complex at promoter plus Co-IP for binding partners, single lab, multiple methods\",\n      \"pmids\": [\"37354982\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NUSAP1 functions as a positive regulator of YAP1 protein stability in gastric cancer cells. NUSAP1 was identified as a novel binding partner of YAP1 by Co-IP, and NUSAP1 depletion reduces YAP1 levels and downstream Hippo target gene transcription (CTGF, CYR61). Cancer-promoting effects of NUSAP1 on cell growth, migration and invasion are largely mediated through YAP1.\",\n      \"method\": \"Co-IP, siRNA knockdown, xenograft model, epistasis rescue (YAP1-dependent phenotype)\",\n      \"journal\": \"Frontiers in oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 / Moderate — Co-IP identifying binding, plus functional epistasis showing YAP1 mediates NUSAP1 oncogenic effects, single lab\",\n      \"pmids\": [\"33489890\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NUSAP1 interacts with CDC20 and CCNA2 (Cyclin A2) by co-immunoprecipitation in osteosarcoma cells. NUSAP1 accelerates cell proliferation and cell cycle progression, and silencing either CDC20 or CCNA2 inhibits NUSAP1-induced proliferation and cell cycle progression.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, CCK-8 and EdU proliferation assays, flow cytometry, xenograft model\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — single Co-IP with functional epistasis by downstream knockdown, single lab\",\n      \"pmids\": [\"34079289\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NUSAP1 depletion in glioblastoma cells leads to downregulation of TOP2A, and silencing either NUSAP1 or TOP2A inhibits GBM cell proliferation and invasion and induces apoptosis. NUSAP1 expression is strongly correlated with TOP2A in glioma datasets, and stable NUSAP1 knockdown reduces tumor growth in an orthotopic xenograft model.\",\n      \"method\": \"RNA sequencing of NUSAP1-stable knockdown cells, siRNA knockdown of NUSAP1 and TOP2A, proliferation/invasion assays, orthotopic xenograft\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — RNA-seq plus functional epistasis (double KD) in multiple cell lines and in vivo model, but no direct binding shown; single lab\",\n      \"pmids\": [\"35532155\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"NUSAP1 expression is regulated by the RB1/E2F1 axis: RB1 knockdown increases NUSAP1 expression, and E2F1 knockdown decreases it. NUSAP1 knockdown in RB1-null or RB1-depleted prostate cancer cells decreases cell proliferation and invasion.\",\n      \"method\": \"Lentiviral RB1 knockdown, transient E2F1 siRNA, RT-qPCR, Western blot, proliferation and invasion assays\",\n      \"journal\": \"The Prostate\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis with two upstream factors (RB1, E2F1) and functional KD phenotype; single lab, multiple methods\",\n      \"pmids\": [\"25585568\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"HOXB2 transcriptionally activates NUSAP1; dual-luciferase reporter and chromatin immunoprecipitation assays confirmed that HOXB2 binds the NUSAP1 promoter. HOXB2 overexpression abrogated the inhibitory effects of NUSAP1 silencing on nephroblastoma cell proliferation and metastasis via the PI3K/Akt signaling pathway.\",\n      \"method\": \"Dual-luciferase reporter assay, ChIP assay, JASPAR bioinformatics, Western blot for PI3K/Akt pathway proteins, siRNA and overexpression transfection\",\n      \"journal\": \"Molecular medicine reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP plus luciferase reporter verifying direct transcriptional regulation; functional epistasis with pathway readout; single lab\",\n      \"pmids\": [\"35485274\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Transcription factor E2F8 directly binds the NUSAP1 promoter (confirmed by ChIP and dual-luciferase reporter) and regulates NUSAP1 transcription. The E2F8/NUSAP1 axis promotes cisplatin resistance in hepatocellular carcinoma by reducing DNA damage (γ-H2AX), with NUSAP1 knockdown sensitizing cells to cisplatin.\",\n      \"method\": \"ChIP assay, dual-luciferase reporter, siRNA knockdown, CCK-8, flow cytometry, comet assay, Western blot\",\n      \"journal\": \"International journal of toxicology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP and luciferase reporter confirm direct transcriptional regulation; functional downstream assays; single lab\",\n      \"pmids\": [\"37331996\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"PRMT1 catalyzes asymmetric dimethylation of NUSAP1 at R418 and R422 (ADMA). The R422 methylation site is critical for NUSAP1 function: methylated NUSAP1 interacts with the PEST domain of Notch2, inhibiting Notch2 ubiquitination and stabilizing Notch2 protein to activate Notch2 signaling. Inhibition of PRMT1 or R422 mutation abolishes NUSAP1-mediated Notch2 stabilization and 5-FU resistance in gastric cancer.\",\n      \"method\": \"Proteomic analysis, site-directed mutagenesis, Co-IP, ubiquitination assay, Western blot, functional resistance assays\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — mutagenesis of specific methylation sites with Co-IP and ubiquitination assay demonstrating mechanism; single lab, multiple methods\",\n      \"pmids\": [\"40393971\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"O-GlcNAcylation (mediated by OGT) enhances NUSAP1 protein stability in bladder cancer cells. Pharmacological upregulation of O-GlcNAc increases NUSAP1 protein levels and stability, and OGT knockdown reduces tumor growth, while NUSAP1 overexpression impairs this effect, placing NUSAP1 downstream of OGT-mediated O-GlcNAcylation.\",\n      \"method\": \"Western blot, pharmacological O-GlcNAc modulation (GlcNAc, PuGNAc), OGT siRNA, CCK-8, flow cytometry, xenograft\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, no direct demonstration of NUSAP1 O-GlcNAcylation site; stability inferred from protein levels after OGT manipulation\",\n      \"pmids\": [\"33488099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ANKRD22 regulates NUSAP1 expression, and NUSAP1 overexpression reverses the inhibitory effects of ANKRD22 knockdown on breast cancer cell proliferation, invasion, and EMT. NUSAP1 mediates ANKRD22 activation of the Wnt/β-catenin signaling pathway.\",\n      \"method\": \"siRNA knockdown, overexpression rescue assay, Western blot for β-catenin pathway, BrdU, colony formation, Transwell assays\",\n      \"journal\": \"Bosnian journal of basic medical sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — epistasis rescue experiment placing NUSAP1 downstream of ANKRD22, single lab, no direct binding shown\",\n      \"pmids\": [\"32651974\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NUSAP1 promotes early recurrence of hepatocellular carcinoma by stimulating STAT3 nuclear translocation and activation through RACK1. RNA sequencing showed enrichment of cancer stemness and STAT3 pathway gene sets upon NUSAP1 overexpression.\",\n      \"method\": \"RNA sequencing, gain/loss-of-function experiments, in vivo recurrence mouse model, pathway analysis\",\n      \"journal\": \"Cancer science\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — STAT3 pathway activation proposed via RACK1 linkage from RNA-seq; mechanistic detail sparse in abstract; single lab\",\n      \"pmids\": [\"36106345\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NUSAP1 functions as a tumor suppressor in triple-negative breast cancer by recruiting the transcriptional repressor DAXX via its microtubule-associated domain (MAD) to bridge HIFα. This complex recruits SETDB1 methyltransferase and deposits H3K9me3 repressive marks on hypoxia response elements (HREs), attenuating HIF transcriptional activity and expression of HIF target genes.\",\n      \"method\": \"NUSAP1 depletion/overexpression, in vitro and in vivo invasion/proliferation assays, Co-IP (NUSAP1-DAXX-HIFα), ChIP for H3K9me3 at HREs, engineered MAD domain rescue\",\n      \"journal\": \"Advanced science (Weinheim, Baden-Wurttemberg, Germany)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating ternary complex plus ChIP showing H3K9me3 deposition at HREs; functional domain-engineering rescue; single lab, multiple orthogonal methods\",\n      \"pmids\": [\"41178464\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NUSAP1 interacts with ANXA2 (Annexin A2) and protects it from ubiquitin-dependent protein degradation, thereby increasing ANXA2 stability. NUSAP1 silencing increases GC radiosensitivity by impairing DNA damage repair after irradiation.\",\n      \"method\": \"Immunoprecipitation, mass spectrometry, ubiquitination assay, colony formation, flow cytometry, comet assay, xenograft\",\n      \"journal\": \"Journal of cancer research and clinical oncology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — IP-MS identifies binding partner, ubiquitination assay demonstrates mechanism of stabilization, functional radiosensitivity readout; single lab\",\n      \"pmids\": [\"39212774\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NUSAP1 interacts with SHCBP1 and inhibits JAK2/STAT3 phosphorylation by blocking SHCBP1 in hepatocellular carcinoma cells, promoting dendritic cell generation from co-cultured PBMCs.\",\n      \"method\": \"Co-immunoprecipitation, Western blot for phospho-JAK2/STAT3, PBMC co-culture with HCC cells, flow cytometry for CD1a/CD86\",\n      \"journal\": \"Journal of immunotherapy\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single Co-IP with pathway readout; mechanistic detail limited in abstract; single lab\",\n      \"pmids\": [\"38980111\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NUSAP1 directly interacts with KIF2C (a kinesin-13 microtubule depolymerase) and Aurora kinase A (AURKA). AURKA-mediated phosphorylation of NUSAP1 modulates NUSAP1's binding affinity to KIF2C, balancing spindle microtubule stability and depolymerization. Truncating NUSAP1 mutations (Y403* and Q405*) impair NUSAP1-KIF2C interaction. NUSAP1 rescues cell cycle arrest and apoptosis caused by KIF2C overexpression in NUSAP1-KO cells, but mutant truncated NUSAP1 fails this rescue.\",\n      \"method\": \"Co-IP (NUSAP1-KIF2C, NUSAP1-AURKA), phosphorylation assay, NUSAP1-KO HEK293T rescue experiment, mouse microcephaly model, live cell imaging\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating direct interactions, phosphorylation assay, domain-disrupting mutation rescue experiment; preprint, single lab, multiple methods\",\n      \"pmids\": [\"bio_10.1101_2025.05.16.654427\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"A recurrent de novo heterozygous nonsense variant (p.Tyr403Ter) in NUSAP1 causes microcephaly, severe developmental delay, and epilepsy. The mutant transcript escapes nonsense-mediated decay, suggesting dominant-negative or gain-of-function mechanism rather than haploinsufficiency.\",\n      \"method\": \"Exome sequencing, NMD escape assay (transcript analysis), single-cell RNA-seq of post-mortem brain\",\n      \"journal\": \"Clinical genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — NMD escape experimentally demonstrated; mechanism (dominant-negative vs. GOF) not yet resolved; single lab\",\n      \"pmids\": [\"37005340\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In zebrafish, nusap1 morpholino knockdown impairs neural crest cell migration, alters expression of neural crest markers crestin and sox9b, and causes apoptosis in retina and hindbrain. These phenotypes are rescued by co-injection of nusap1 mRNA. This establishes an in vivo role for Nusap1 in cell migration and neural crest morphogenesis.\",\n      \"method\": \"Antisense morpholino knockdown in zebrafish, in situ hybridization for neural crest markers, mRNA rescue injection\",\n      \"journal\": \"Protein & cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — morpholino KD with mRNA rescue (gold standard in zebrafish), multiple marker readouts; single lab\",\n      \"pmids\": [\"21203972\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"In mouse oocytes, NUSAP1 localizes to distinct granular aggregates near spindle poles during Pro-MI, metaphase I, and anaphase I. NUSAP1 depletion causes chromosome misalignment, increased aneuploidy, abnormal spindle assembly with decreased spindle pole width, and attenuation of cortical F-actin. Mass spectrometry of NUSAP1 interactomes showed significant enrichment for RNA-binding partners, suggesting NUSAP1 influences P-body dynamics and mRNA homeostasis.\",\n      \"method\": \"Protein expression profiling (proteomics), siRNA depletion, immunofluorescence for localization, aneuploidy measurement, RNA-seq, mass spectrometry interactome\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct localization by imaging with functional depletion phenotype and MS interactome; single lab, multiple methods\",\n      \"pmids\": [\"37992207\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NUSAP1 interacts with NUSAP1-IP3R (inositol 1,4,5-trisphosphate receptor); NUSAP1 governs IP3R phosphorylation status to regulate cytosolic Ca2+ homeostasis and ER stress. NUSAP1 knockdown increases cytosolic Ca2+, p-IP3R, and ER stress, promoting mitophagy and apoptosis in endometrial carcinoma cells. IP3R silencing reduces Ca2+ and ER stress without affecting NUSAP1 expression, but NUSAP1 overexpression combined with IP3R knockdown amplified these effects synergistically.\",\n      \"method\": \"Co-IP (NUSAP1-IP3R interaction), Fluo-4 AM Ca2+ measurement, ER fluorescence probe imaging, flow cytometry, Western blot, siRNA knockdown and overexpression\",\n      \"journal\": \"ACS omega\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — Co-IP demonstrates interaction; IP3R phosphorylation mechanism inferred from pharmacological and genetic epistasis; single lab, no in vitro kinase/phosphatase assay\",\n      \"pmids\": [\"41585655\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"HBV core protein (HBC) enhances WDR46 protein stability by blocking TRIM25-mediated ubiquitination of WDR46. WDR46 then facilitates c-Myc recruitment to the NUSAP1 promoter by enhancing the WDR46-c-Myc interaction, thereby transcriptionally upregulating NUSAP1 to promote HCC cell growth and migration.\",\n      \"method\": \"Co-immunoprecipitation, ChIP, Western blot, ubiquitination assay, functional growth and migration assays, xenograft model\",\n      \"journal\": \"Hepatology communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — ChIP confirms c-Myc recruitment to NUSAP1 promoter; Co-IP and ubiquitination assay delineate upstream WDR46 stabilization mechanism; single lab, multiple methods\",\n      \"pmids\": [\"40366140\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NUSAP1 promotes lung adenocarcinoma cell proliferation, and its upregulation is mediated by estrogen via ERβ activation. Pharmacological targeting of the ERβ/NUSAP1 axis with fulvestrant (ERβ antagonist) or entinostat (identified as a novel NUSAP1 inhibitor) suppresses LUAD growth in vitro and in vivo.\",\n      \"method\": \"Bioinformatics, in vitro proliferation assays with estrogen treatment, ERβ activation/inhibition, entinostat treatment, in vivo xenograft\",\n      \"journal\": \"International journal of biological sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — regulatory mechanism (estrogen-ERβ-NUSAP1) demonstrated pharmacologically; direct binding/transcriptional mechanism not fully delineated in abstract; single lab\",\n      \"pmids\": [\"39430250\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NUSAP1 is a microtubule-associated protein that localizes to the mitotic spindle and plays key roles in spindle assembly, chromosome segregation, and DNA damage response: it interacts with the RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex via its SAP domain (which contains two functional SUMOylation consensus sites regulated by RANBP2), promotes ATR stability via SAP domain-mediated sumoylation, regulates BRCA1 protein levels to support DSB repair and centrosome duplication, suppresses R-loop accumulation through interaction with RNA-binding protein ILF2, and is phosphorylated by AURKA to modulate its interaction with KIF2C for spindle microtubule balance; in cancer contexts NUSAP1 forms a transcriptional complex with c-Myc and HIF-1α to drive LDHA expression, stabilizes YAP1 and ANXA2 proteins by suppressing their ubiquitination, and is itself transcriptionally controlled by the RB1/E2F1 axis and by HOXB2/E2F8; PRMT1-mediated dimethylation at R422 enables NUSAP1 to stabilize Notch2 via PEST domain interaction, while in triple-negative breast cancer NUSAP1 acts as a tumor suppressor by recruiting DAXX to HIFα and depositing H3K9me3 repressive marks at hypoxia response elements.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NUSAP1 is a microtubule-associated protein that localizes to dynamic spindle microtubules in a chromosome-centric pattern during metaphase and anaphase, where it organizes spindle architecture and chromosome segregation through SUMO-pathway and kinesin interactions [#0, #19]. Through its SAP domain it engages the RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex, and two consensus SUMOylation sites within its DNA- and microtubule-binding domains are modified in a RANBP2-dependent manner, each site selectively controlling NUSAP1 localization and dictating distinct mitotic outcomes [#0, #1]. NUSAP1 directly binds KIF2C and Aurora kinase A; AURKA-mediated phosphorylation tunes NUSAP1-KIF2C affinity to balance spindle microtubule stability against depolymerization, and truncating mutations (Y403*, Q405*) that disrupt this interaction abolish its rescue activity [#19]. Beyond mitosis, NUSAP1 supports genome stability: it sustains BRCA1 protein levels and BRCA1 focus formation to enable homologous-recombination and single-strand-annealing repair and proper centrosome duplication [#4], and it acts upstream of the RNA-binding protein ILF2 (also interacting with DHX9) to suppress R-loop accumulation and DNA damage [#3]. Across cancers, NUSAP1 is deployed as an oncogenic protein-stabilizing and transcription-regulating hub—forming a c-Myc/HIF-1\\u03b1 complex at the LDHA promoter to drive glycolysis [#5], stabilizing partner proteins including ATR, YAP1, ANXA2 and Notch2 by antagonizing their ubiquitin-dependent degradation [#2, #6, #17, #12], and is itself transcriptionally controlled by the RB1/E2F1 and E2F8 axes [#9, #11]. In triple-negative breast cancer, by contrast, NUSAP1 acts as a tumor suppressor, recruiting DAXX via its microtubule-associated domain to bridge HIF\\u03b1 and deposit SETDB1-dependent H3K9me3 marks on hypoxia response elements to repress HIF target genes [#16]. A recurrent de novo nonsense variant (p.Tyr403Ter) that escapes nonsense-mediated decay causes microcephaly, severe developmental delay, and epilepsy [#20], consistent with conserved developmental roles seen in zebrafish neural crest migration [#21] and mouse oocyte spindle assembly [#22].\",\n  \"teleology\": [\n    {\n      \"year\": 2014,\n      \"claim\": \"Established that NUSAP1 contributes to genome maintenance beyond its mitotic role by linking it to double-strand break repair and centrosome integrity.\",\n      \"evidence\": \"siRNA depletion with HR/SSA repair assays, BRCA1 focus imaging, and plasmid rescue in human cells\",\n      \"pmids\": [\"24521615\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which NUSAP1 controls BRCA1 protein level not defined\", \"No direct NUSAP1-BRCA1 binding shown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Placed NUSAP1 transcription under the RB1/E2F1 axis, explaining its cell-cycle-coupled expression and elevation in RB1-deficient cancer.\",\n      \"evidence\": \"Lentiviral RB1 and siRNA E2F1 knockdown with RT-qPCR, Western blot, and proliferation/invasion assays in prostate cancer cells\",\n      \"pmids\": [\"25585568\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct E2F1 binding to the NUSAP1 promoter not demonstrated here\", \"Does not address post-translational regulation\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Identified the molecular basis of NUSAP1's mitotic function as a SUMO-pathway cofactor by mapping its interaction with the RanBP2-RanGAP1-UBC9 complex.\",\n      \"evidence\": \"Endogenous reciprocal Co-IP with mass spectrometry, live-cell imaging, and Taxol-sensitivity phenotyping in human cells\",\n      \"pmids\": [\"28900032\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific SUMO substrates recognized via the SAP domain not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Demonstrated an in vivo developmental requirement for NUSAP1 in cell migration and neural crest morphogenesis.\",\n      \"evidence\": \"Zebrafish morpholino knockdown with neural crest marker in situ hybridization and mRNA rescue\",\n      \"pmids\": [\"21203972\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism linking NUSAP1 to neural crest migration unresolved\", \"Morpholino specificity limitations\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Provided a substrate-level mechanism for SAP-domain function by showing NUSAP1 promotes ATR sumoylation to antagonize its proteolysis and confer chemoresistance.\",\n      \"evidence\": \"Co-IP, sumoylation and ubiquitination assays, SAP-domain mutation, and viability assays in glioblastoma cells\",\n      \"pmids\": [\"32317623\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct SUMO E3 activity vs cofactor role not separated\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Extended NUSAP1's protein-stabilizing role to cancer signaling hubs YAP1 and cell-cycle regulators CDC20/CCNA2 driving proliferation.\",\n      \"evidence\": \"Co-IP, siRNA knockdown, epistasis rescue, and xenografts in gastric cancer and osteosarcoma cells\",\n      \"pmids\": [\"33489890\", \"34079289\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether NUSAP1 stabilizes YAP1 by blocking ubiquitination not directly tested\", \"Direct binding surfaces undefined\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Mapped additional transcriptional inputs (HOXB2) and downstream effectors (TOP2A) that integrate NUSAP1 into proliferative cancer programs.\",\n      \"evidence\": \"ChIP and dual-luciferase reporter for HOXB2, RNA-seq plus double knockdown for TOP2A, with xenograft models\",\n      \"pmids\": [\"35485274\", \"35532155\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"TOP2A regulation shown without direct binding\", \"Tissue specificity of these axes unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Resolved the site-specific logic of NUSAP1 SUMOylation, showing two RANBP2-dependent sites independently govern localization and daughter-cell fate.\",\n      \"evidence\": \"Site-directed mutagenesis, proximity ligation with SUMO2/3, and mitotic phenotyping in human cells\",\n      \"pmids\": [\"37947624\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Downstream effectors of each modified site not identified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Connected NUSAP1 to RNA metabolism and R-loop suppression by placing it upstream of ILF2/DHX9 in the DNA damage response.\",\n      \"evidence\": \"AP-MS interactome, reciprocal Co-IP, confocal colocalization, domain mapping, and R-loop/DNA damage assays after camptothecin\",\n      \"pmids\": [\"37047232\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How NUSAP1 mechanistically enables ILF2 R-loop suppression unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Defined a metabolic-transcriptional role in which NUSAP1 joins c-Myc and HIF-1\\u03b1 at the LDHA promoter, reinforced by lactylation-driven stabilization.\",\n      \"evidence\": \"ChIP-seq, Co-IP, RNA-seq, single-cell RNA-seq, and xenografts\",\n      \"pmids\": [\"37354982\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Lactylation site on NUSAP1 not mapped\", \"Direct DNA contact by NUSAP1 vs partners unclear\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Identified E2F8 as an additional direct transcriptional activator of NUSAP1 mediating cisplatin resistance.\",\n      \"evidence\": \"ChIP, dual-luciferase reporter, knockdown, comet assay, and \\u03b3-H2AX analysis in hepatocellular carcinoma\",\n      \"pmids\": [\"37331996\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Relationship to RB1/E2F1 axis not integrated\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Confirmed conserved meiotic spindle and RNA-associated functions in mammalian oocytes, reinforcing the link between NUSAP1 and RNA-binding partners.\",\n      \"evidence\": \"Proteomics, siRNA depletion, immunofluorescence localization, aneuploidy scoring, and MS interactome in mouse oocytes\",\n      \"pmids\": [\"37992207\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct role in P-body dynamics inferred, not demonstrated\", \"Specific RNA targets unidentified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Linked NUSAP1 to human neurodevelopmental disease through a recurrent de novo nonsense variant acting via a non-haploinsufficiency mechanism.\",\n      \"evidence\": \"Exome sequencing, NMD escape transcript assay, and single-cell RNA-seq of post-mortem brain\",\n      \"pmids\": [\"37005340\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Dominant-negative vs gain-of-function mechanism unresolved\", \"Single family/lab cohort\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Extended the protein-stabilization paradigm to ANXA2, tying NUSAP1 to radioresistance via DNA-damage repair.\",\n      \"evidence\": \"IP-MS, ubiquitination assay, comet assay, and xenografts in gastric cancer\",\n      \"pmids\": [\"39212774\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"E3 ligase antagonized by NUSAP1 not identified\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Established the direct AURKA-NUSAP1-KIF2C axis that balances spindle microtubule dynamics and tied disease-associated truncations to its disruption.\",\n      \"evidence\": \"Co-IP, phosphorylation assay, NUSAP1-KO rescue, mouse microcephaly model, and live imaging (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.05.16.654427\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint, single lab\", \"Phosphosites on NUSAP1 not fully mapped\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Revealed a context-dependent tumor-suppressor function in which NUSAP1 represses HIF transcriptional output via DAXX/SETDB1-mediated H3K9me3 deposition.\",\n      \"evidence\": \"Co-IP of NUSAP1-DAXX-HIF\\u03b1, ChIP for H3K9me3 at HREs, and MAD-domain rescue in triple-negative breast cancer\",\n      \"pmids\": [\"41178464\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Determinants of oncogenic vs tumor-suppressor switching unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Added a PRMT1-dependent methylation layer (R418/R422) enabling NUSAP1 to stabilize Notch2 and drive 5-FU resistance.\",\n      \"evidence\": \"Proteomics, site-directed mutagenesis, Co-IP, and ubiquitination assays in gastric cancer\",\n      \"pmids\": [\"40393971\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of methyl-dependent PEST-domain binding undefined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"What governs the switch between NUSAP1's oncogenic protein-stabilizing/transcriptional roles and its tumor-suppressive HIF-repressive role, and how its mitotic, DNA-repair, and disease-causing functions are mechanistically unified, remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying biochemical model across contexts\", \"Structural basis of SAP/MAD domain partner selection unknown\", \"Direct enzymatic vs adaptor role in SUMO and ubiquitin pathways not separated\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 19]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 16, 19]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [5, 16]},\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [2, 17, 12]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [1]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 22]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [5, 16]},\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1, 19, 7]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [4, 3, 17]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [5, 16]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [2, 6, 17, 12]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [20]}\n    ],\n    \"complexes\": [\n      \"RanBP2-RanGAP1-UBC9 SUMO E3 ligase complex\",\n      \"c-Myc/HIF-1\\u03b1/NUSAP1 transcriptional complex at LDHA promoter\",\n      \"NUSAP1-DAXX-HIF\\u03b1 repressive complex\"\n    ],\n    \"partners\": [\n      \"RANBP2\",\n      \"KIF2C\",\n      \"AURKA\",\n      \"ILF2\",\n      \"DHX9\",\n      \"YAP1\",\n      \"ANXA2\",\n      \"DAXX\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}