{"gene":"HASPIN","run_date":"2026-06-10T01:55:21","timeline":{"discoveries":[{"year":2005,"finding":"Haspin phosphorylates histone H3 at Thr3 in vitro and is required for H3T3 phosphorylation in mitotic cells; depletion by RNAi causes metaphase chromosome misalignment and Haspin associates with chromosomes, centrosomes, and spindle during mitosis.","method":"In vitro kinase assay, RNAi depletion, immunostaining with phospho-specific antibodies","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — in vitro kinase assay establishing substrate, RNAi loss-of-function with clear cellular phenotype, replicated in subsequent studies","pmids":["15681610"],"is_preprint":false},{"year":2006,"finding":"Haspin depletion disrupts centromeric cohesin binding and sister chromatid cohesion in mitosis, causing chromosome misalignment and spindle assembly checkpoint activation; overexpression stabilizes arm cohesion, indicating Haspin is required to maintain centromeric cohesion.","method":"RNAi depletion, immunostaining for cohesin and H3T3ph, live-cell imaging","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — RNAi loss-of-function with specific cohesin phenotype, gain-of-function overexpression, replicated by subsequent studies","pmids":["17084365"],"is_preprint":false},{"year":2009,"finding":"Crystal structure of the human Haspin kinase domain reveals a constitutively active conformation stabilized by haspin-specific inserts, an atypical activation segment without phosphorylatable residues, and that His651 in the catalytic loop plays a direct role in ATP recognition (His651Ala mutant is inactive). Increasing H3K4 methylation strongly decreases substrate recognition.","method":"X-ray crystallography, active-site mutagenesis (His651Ala), enzyme kinetic assays","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — crystal structure with functional mutagenesis and enzyme kinetics, confirmed by a second independent structure paper (PMID:19918049) in the same year","pmids":["19918057","19918049"],"is_preprint":false},{"year":2009,"finding":"Crystal structure of human Haspin kinase domain (2.15 Å) shows the αC helix adopts its active conformation stabilized by a helical insertion and an unprecedented activation segment conformation, explaining constitutive activity without external activation loop phosphorylation.","method":"X-ray crystallography (2.15 Å resolution)","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 / Strong — high-resolution crystal structure, independently confirms findings of PMID:19918057","pmids":["19918049"],"is_preprint":false},{"year":2010,"finding":"Haspin-generated H3T3 phosphorylation provides a direct chromatin docking site for the CPC subunit Survivin at centromeres; Survivin binds directly to H3T3ph (pulldown), and a non-binding Survivin-D70A/D71A mutant fails to support centromeric CPC concentration, diminishing Aurora B centromere functions including MCAK localization and spindle checkpoint response.","method":"Direct binding assay (Survivin pulldown with H3T3ph peptide), Survivin point mutant rescue, antibody microinjection, RNAi","journal":"Science","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — direct binding assay identifying Survivin as H3T3ph reader, mutagenesis of binding interface, replicated across multiple labs","pmids":["20705812"],"is_preprint":false},{"year":2011,"finding":"Aurora B phosphorylates Haspin to promote H3T3ph generation, establishing a positive CPC–Haspin–H3T3ph feedback loop at centromeres; Aurora B kinase activity is required for normal chromosomal CPC localization. The Bub1–shugoshin–CPC pathway provides a centromere-specific boost to this feedback loop.","method":"Kinase assays, RNAi, Aurora B inhibitor treatment, live-cell imaging","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal kinase relationship demonstrated biochemically and in cells, multiple orthogonal methods, replicated by other studies","pmids":["21658950"],"is_preprint":false},{"year":2013,"finding":"A conserved basic segment in Haspin's N-terminus autoinhibits kinase activity during interphase; Cdk1 phosphorylation of the N-terminus recruits Plk1/Plx1, which further phosphorylates multiple N-terminal sites to neutralize autoinhibition and activate Haspin in M phase.","method":"In vitro kinase assays, Xenopus egg extract experiments, mutagenesis of autoinhibitory segment and phosphorylation sites","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 / Strong — biochemical reconstitution in egg extracts, mutagenesis of regulatory sites, multiple orthogonal methods","pmids":["24184212"],"is_preprint":false},{"year":2014,"finding":"Plk1 binds to Haspin in a Cdk1-phosphorylation-dependent manner and is required for initial Haspin phosphorylation and H3T3ph in prophase, positively regulating CPC recruitment at mitotic entry.","method":"Co-immunoprecipitation, Plk1 inhibitor treatment, phospho-Haspin immunostaining","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal interaction confirmed, consistent with Xenopus data from PMID:24184212, two independent studies","pmids":["24413556"],"is_preprint":false},{"year":2014,"finding":"Co-crystal structure of Haspin with the histone H3 tail reveals a unique bent substrate binding mode positioning H3 residues Arg2 and Lys4 into acidic binding pockets adjacent to the phosphorylated Thr3, explaining how H3K4 methylation reduces Haspin activity by steric/charge interference.","method":"X-ray co-crystallography, mass spectrometry-based phosphoproteomics, in vitro kinase assays","journal":"Molecular & cellular proteomics : MCP","confidence":"High","confidence_rationale":"Tier 1 / Moderate — co-crystal structure with substrate peptide, structural basis for methylation-dependent regulation, single lab but multiple orthogonal methods","pmids":["24732914"],"is_preprint":false},{"year":2014,"finding":"Haspin phosphorylates macroH2A at Ser137 in addition to H3T3; the Ser137 site is in a basic stretch required to stabilize extranucleosomal DNA, suggesting phosphorylation regulates macroH2A–DNA interactions.","method":"Mass spectrometry-based phosphoproteomics, in vitro kinase assays, consensus motif analysis","journal":"Molecular & cellular proteomics : MCP","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — identified by MS and validated in vitro, single lab, functional consequence inferred but not fully proven","pmids":["24732914"],"is_preprint":false},{"year":2016,"finding":"SUMOylation of DNA topoisomerase IIα CTD is required for Haspin localization at centromeres and H3T3ph; SUMOylated TOP2A CTD directly interacts with Haspin through SUMO-interaction motifs to regulate CPC recruitment in mitosis, established using Xenopus egg extracts.","method":"Xenopus egg extract biochemistry, SUMO site mutagenesis, co-immunoprecipitation","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — biochemical reconstitution in egg extracts, mutagenesis of SUMO sites, direct interaction demonstrated","pmids":["27325792"],"is_preprint":false},{"year":2017,"finding":"Haspin binds the cohesin regulatory subunit Pds5B through a conserved YGA/R motif in its non-catalytic N-terminus (similar to Wapl's YSR motif); disruption of this interaction causes weakened centromeric cohesion and premature chromatid separation, rescued by prevention of Wapl-mediated cohesin removal.","method":"Co-immunoprecipitation, CRISPR knockout, domain mutant rescue experiments","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 / Moderate — reciprocal co-IP, genetic rescue with specific motif mutant, multiple orthogonal approaches","pmids":["28343965"],"is_preprint":false},{"year":2017,"finding":"The Haspin kinase domain directly binds and phosphorylates the YSR motif of Wapl, inhibiting Wapl–Pds5B interaction and preventing Wapl-dependent cohesin release from centromeres; phospho-mimetic mutation in Wapl-YSR prevents Wapl from releasing cohesin.","method":"In vitro kinase assay, co-immunoprecipitation, phospho-mimetic mutagenesis, centromere targeting assays","journal":"EMBO reports","confidence":"High","confidence_rationale":"Tier 1–2 / Moderate — in vitro kinase assay showing direct phosphorylation of Wapl-YSR, phospho-mimetic mutagenesis demonstrating functional consequence, complementary to PMID:28343965","pmids":["29138236"],"is_preprint":false},{"year":2017,"finding":"Aurora-A phosphorylates Haspin at multiple N-terminal sites in late G2/nuclear phase to promote H3T3ph and CPC centromere recruitment before spindle assembly, independently of Plk1; Aurora-A also facilitates association of Aurora-B with Haspin and Plk1.","method":"Aurora-A knockout/inhibition/nuclear-import blockade, phosphorylation site mapping, centromere localization assays in cells","journal":"Cell discovery","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — single lab, multiple cell-based methods but limited in vitro reconstitution","pmids":["28101375"],"is_preprint":false},{"year":2009,"finding":"RNAi depletion of Haspin (along with Sgo1 and Scc1) causes generation of multiple acentriolar centrosome-like foci and spindle disruption only when chromatid cohesion is lost; these effects require kinesin-5 activity and microtubule dynamics, indicating spindle-pole integrity depends indirectly on chromosome cohesion.","method":"RNAi, live-cell imaging, topoisomerase II inhibition rescue, kinesin-5 inhibitor","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — live imaging with specific genetic and pharmacological perturbations, but spindle pole disruption is shown to be indirect","pmids":["19910498"],"is_preprint":false},{"year":2012,"finding":"Haspin inhibitors (and anti-H3T3ph antibody microinjection) decrease centromeric Aurora B and its substrates (CENP-A, Hec1), compromise metaphase chromosome alignment and spindle checkpoint signaling; retargeting Aurora B to centromeres bypasses the Haspin requirement, confirming the Haspin→H3T3ph→CPC centromere recruitment pathway.","method":"Small-molecule Haspin inhibitors, antibody microinjection, Aurora B retargeting, phosphosubstrate immunostaining","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple inhibitor tools plus antibody injection plus epistatic rescue (Aurora B retargeting), independently replicated in two simultaneous papers","pmids":["23071152","23071153"],"is_preprint":false},{"year":2011,"finding":"Methylation at H3K4 abolishes Haspin-mediated H3T3 phosphorylation; dimethylation at H3R2 also substantially reduces Haspin activity on H3T3, establishing combinatorial histone modification control of Haspin substrate recognition.","method":"In vitro kinase assays with variously modified H3 peptides","journal":"Bioorganic & medicinal chemistry","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — in vitro kinase assays with modified peptides, single lab, consistent with structural data","pmids":["21397507"],"is_preprint":false},{"year":1999,"finding":"Haspin (mouse) has intrinsic Ser/Thr kinase activity; it localizes exclusively to nuclei; autophosphorylation regulates its DNA-binding activity (deletion mutant lacking autophosphorylation binds DNA-cellulose constitutively); ectopic expression without kinase activity causes G1 cell cycle arrest.","method":"In vitro kinase assay, nuclear localization by immunostaining, DNA-cellulose binding assay, transfection with kinase-dead mutant","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — first identification paper with in vitro kinase assay and cell-based localization, single lab","pmids":["10358056"],"is_preprint":false},{"year":2016,"finding":"In mouse oocytes, Haspin is required for Aurora kinase C (AURKC) localization at both chromosomes and acentriolar MTOCs; Haspin inhibition perturbs MTOC clustering into two poles and spindle bipolarity, and AURKC overexpression rescues these defects.","method":"Haspin inhibitor treatment, overexpression rescue, live imaging of AURKC and MTOC markers in mouse oocytes","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — inhibitor with OE rescue, multiple imaging endpoints, single lab","pmids":["27562071"],"is_preprint":false},{"year":2014,"finding":"In mouse oocytes, Haspin-mediated H3T3ph recruits the CPC along chromosome arms (rather than kinetochores), and Haspin inhibition causes improper kinetochore-microtubule attachments and aneuploidy at metaphase II; AURKC rather than AURKB mediates correction of microtubule attachments during meiosis I.","method":"Haspin inhibitor and AURKC inhibitor treatment, chromosome alignment and aneuploidy assays in mouse oocytes","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological loss-of-function with specific phenotypic readouts, single lab","pmids":["25315835"],"is_preprint":false},{"year":2020,"finding":"Either Haspin or Bub1 activity alone is sufficient to recruit Aurora B to a centromeric locus; combined inhibition of both Haspin and Bub1 fully abolishes Aurora B centromere accumulation and impairs correction of erroneous KT-MT attachments but does not compromise the mitotic checkpoint or kinetochore Aurora B substrate phosphorylation, arguing against a purely centromere-based spatial model.","method":"Selective kinase inhibitors (Haspin + Bub1), Aurora B substrate phosphorylation assays, chromosome segregation analysis","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 / Moderate — dual inhibitor epistasis with multiple molecular readouts, single lab but rigorous experimental design","pmids":["32027339"],"is_preprint":false},{"year":2020,"finding":"Catalytic inhibition of Topo II leads to its SUMOylation, which triggers H3T3ph via Haspin and recruitment of Aurora B to ectopic chromosomal sites; both Haspin and Aurora B are required for this metaphase checkpoint response.","method":"Topo II catalytic inhibitor, SUMOylation site mutants, Haspin and Aurora B inhibitors, centromere localization assays","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic (SUMOylation mutants) and pharmacological epistasis, single lab","pmids":["31712254"],"is_preprint":false},{"year":2017,"finding":"In Drosophila, Haspin phosphorylates H3T3 both in mitosis and interphase; H3T3ph in interphase localizes to heterochromatin and lamin-enriched euchromatin; Haspin loss compromises insulator activity, reduces nuclear size, and is required for Polycomb-dependent homeotic gene silencing; Haspin associates with the cohesin complex in interphase and mediates Pds5 binding to chromatin.","method":"Haspin mutant Drosophila, ChIP, enhancer-blocking assays, live imaging, co-immunoprecipitation","journal":"PLoS genetics","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic loss-of-function with multiple phenotypic and molecular readouts, single lab","pmids":["32750047"],"is_preprint":false},{"year":2021,"finding":"Haspin phosphorylates H3T3 in quiescent cells and its activity is required for primary cilia length regulation and timely cilia resorption; Haspin activity promotes relocalization of Dido3-HDAC6 to the basal body, and this pathway was confirmed in zebrafish.","method":"Haspin inhibitor in quiescent cells, cilia length measurement, Dido3 overexpression rescue, zebrafish model","journal":"International journal of molecular sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — pharmacological inhibition with specific rescue (Dido3), in vivo zebrafish validation, single lab","pmids":["34299370"],"is_preprint":false},{"year":2022,"finding":"Cryo-EM structure of human Haspin kinase domain bound to a nucleosome shows Haspin contacts only nucleosomal DNA, inserting into a supergroove formed by apposing major grooves of two DNA gyres; key basic residues essential for this DNA binding are required for phosphorylation of nucleosomal H3 and binding to mitotic chromatin.","method":"Cryo-EM structure determination, mutagenesis of DNA-binding residues, nucleosomal H3 phosphorylation assay, mitotic chromatin binding assay","journal":"Nature structural & molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — cryo-EM structure combined with mutagenesis and functional phosphorylation assay, single lab but multiple orthogonal methods","pmids":["39979508"],"is_preprint":false},{"year":2022,"finding":"In vitro kinase assays and CRISPR/Cas9 approaches show VRK1 and VRK2 do not phosphorylate H3T3 or H3S10 in mitosis; Haspin is the kinase specifically required for H3T3ph during mitosis.","method":"In vitro kinase assays, KiPIK screening, RNAi, CRISPR/Cas9","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 1–2 / Moderate — multiple methods including in vitro assay and genetic KO, primarily a negative result for VRK1/2 that positively confirms Haspin's exclusive role","pmids":["35778595"],"is_preprint":false},{"year":2017,"finding":"Haspin phosphorylates TH2A (germ cell-specific H2A variant) at Thr127 at meiotic centromeres; identified by candidate kinase approach and confirmed in vivo using Haspin inhibitor.","method":"Haspin inhibitor in oocytes and spermatocytes, phospho-specific antibody, comparison with TH2A-T127A knock-in mice","journal":"Chromosoma","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — in vivo inhibitor confirms Haspin dependence, knock-in mouse controls, single lab","pmids":["28803373"],"is_preprint":false},{"year":2022,"finding":"Two-hybrid screening from mouse testis identified CENPJ/CPAP, KPNA6/importin alpha 6, and C1QBP/HABP1 as Haspin-interacting proteins; Haspin phosphorylates C1QBP in vitro, suggesting a role in spermatogenesis through C1QBP phosphorylation and potentially centrosome formation.","method":"Yeast two-hybrid, co-immunoprecipitation, in vitro kinase assay","journal":"International journal of molecular sciences","confidence":"Low","confidence_rationale":"Tier 3 / Weak — yeast two-hybrid plus in vitro kinase, single lab, limited functional validation","pmids":["36012324"],"is_preprint":false},{"year":2023,"finding":"During mitosis, Haspin regulates Rho-ROCK activity through the GAP protein ARHGAP11A, and ROCK in turn activates LIMK1 and stabilizes the actin cytoskeleton to support spindle orientation; this pathway is required for correct tissue morphology in 3D cultures.","method":"Haspin inhibition/KO, ARHGAP11A interaction studies, ROCK/LIMK1 activation assays, 3D cell culture morphology","journal":"iScience","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — epistasis established in cells with multiple molecular readouts, single lab, limited biochemical reconstitution","pmids":["37841592"],"is_preprint":false}],"current_model":"Haspin is an atypical, constitutively active Ser/Thr kinase that phosphorylates histone H3 at Thr3 (H3T3ph) during mitosis by binding nucleosomal DNA via a supergroove-insertion mode; this mark is read by the CPC subunit Survivin to recruit Aurora B to inner centromeres, where it regulates kinetochore-microtubule attachments and spindle checkpoint signaling. Haspin activity is cell-cycle controlled through Cdk1-dependent autoinhibition neutralization followed by Plk1- and Aurora A-mediated multi-site N-terminal phosphorylation; Aurora B in turn phosphorylates Haspin in a positive feedback loop. Beyond Aurora B positioning, Haspin protects centromeric cohesion by (1) binding cohesin subunit Pds5B via a YGA/R motif to compete with Wapl, and (2) directly phosphorylating Wapl's YSR motif to prevent Wapl–Pds5B interaction. Haspin also phosphorylates macroH2A at Ser137, TH2A at Thr127 in germ cells, and C1QBP in spermatids, and regulates primary cilia dynamics and spindle orientation through ARHGAP11A-Rho-ROCK-LIMK1 in interphase cells."},"narrative":{"mechanistic_narrative":"Haspin is an atypical Ser/Thr kinase that governs the spatial organization of mitotic chromatin signaling, principally by phosphorylating histone H3 at Thr3 (H3T3ph) during mitosis [PMID:15681610]. Crystal structures show that Haspin is constitutively active: haspin-specific helical inserts and an unprecedented activation-segment conformation hold the αC helix and catalytic apparatus in an active state without canonical activation-loop phosphorylation, with His651 directly mediating ATP recognition [PMID:19918057, PMID:19918049]. Substrate engagement is highly selective—co-crystal and peptide studies reveal a bent H3-tail binding mode that positions Arg2 and Lys4 in acidic pockets, so that H3K4 methylation and H3R2 dimethylation sterically and electrostatically suppress phosphorylation [PMID:24732914, PMID:21397507]—and a cryo-EM nucleosome complex shows Haspin contacts only nucleosomal DNA by inserting into a supergroove between two DNA gyres, a mode required for phosphorylation of chromatin-embedded H3 [PMID:39979508]. The H3T3ph mark is read directly by the chromosomal passenger complex (CPC) subunit Survivin, concentrating Aurora B at inner centromeres to control MCAK localization, kinetochore-microtubule attachments, and the spindle checkpoint [PMID:20705812, PMID:23071152, PMID:23071153]. Haspin activity is cell-cycle gated: an autoinhibitory N-terminal basic segment is relieved when Cdk1 phosphorylation recruits Plk1 for multi-site activating phosphorylation at mitotic entry, while Aurora-A primes the pathway in late G2 and Aurora B feeds back positively onto Haspin [PMID:21658950, PMID:24184212, PMID:24413556, PMID:28101375]. In parallel with Aurora B positioning, Haspin protects centromeric cohesion both by binding the cohesin regulator Pds5B through a conserved YGA/R motif and by directly phosphorylating the Wapl YSR motif to block Wapl-mediated cohesin release [PMID:17084365, PMID:28343965, PMID:29138236]. Haspin centromere recruitment depends on SUMOylated topoisomerase IIα, linking Topo II catalytic state to H3T3ph-based checkpoint responses [PMID:27325792, PMID:31712254]. Beyond mitosis, Haspin functions in meiosis to recruit Aurora kinase C in oocytes [PMID:27562071, PMID:25315835], in primary cilia length and resorption [PMID:34299370], and in spindle orientation through an ARHGAP11A-Rho-ROCK-LIMK1 axis [PMID:37841592].","teleology":[{"year":1999,"claim":"Establishing that Haspin was a genuine nuclear Ser/Thr kinase whose own autophosphorylation modulated DNA binding set the foundation for treating it as a chromatin-associated enzyme rather than an orphan sequence.","evidence":"In vitro kinase assay, nuclear immunostaining, DNA-cellulose binding, and kinase-dead transfection in mouse cells","pmids":["10358056"],"confidence":"Medium","gaps":["No physiological substrate identified at this stage","Mechanism linking autophosphorylation to DNA binding not structurally resolved","Cell-cycle regulation of activity unaddressed"]},{"year":2005,"claim":"Identifying H3 Thr3 as Haspin's mitotic substrate answered what the kinase does on chromatin and tied it to chromosome alignment.","evidence":"In vitro kinase assay, RNAi depletion, and phospho-specific immunostaining in mitotic cells","pmids":["15681610"],"confidence":"High","gaps":["Downstream reader of H3T3ph unknown","Structural basis of substrate specificity unresolved","How misalignment arises mechanistically not defined"]},{"year":2006,"claim":"Demonstrating that Haspin maintains centromeric cohesin and sister chromatid cohesion revealed a second, distinct mitotic role beyond histone marking.","evidence":"RNAi depletion, cohesin immunostaining, overexpression, and live-cell imaging","pmids":["17084365"],"confidence":"High","gaps":["Molecular link between Haspin and cohesin retention not defined","Whether this required kinase activity or H3T3ph unresolved"]},{"year":2009,"claim":"Crystal structures explained how Haspin is constitutively active without activation-loop phosphorylation, answering why an atypical kinase still adopts a productive conformation.","evidence":"X-ray crystallography of the kinase domain plus His651Ala mutagenesis and enzyme kinetics, with an independent confirmatory structure","pmids":["19918057","19918049"],"confidence":"High","gaps":["How cell-cycle regulation overrides constitutive activity unexplained","Nucleosomal substrate engagement not captured"]},{"year":2010,"claim":"Identifying Survivin as a direct reader of H3T3ph closed the loop between Haspin's mark and CPC/Aurora B centromere recruitment.","evidence":"Survivin pulldown with H3T3ph peptide, non-binding D70A/D71A mutant rescue, antibody microinjection, and RNAi","pmids":["20705812"],"confidence":"High","gaps":["Quantitative contribution relative to other CPC targeting pathways unresolved","Spatial dynamics of recruitment not addressed"]},{"year":2011,"claim":"Showing Aurora B phosphorylates Haspin and that H3K4/H3R2 methylation blocks H3T3ph defined both a positive feedback loop and combinatorial histone control of substrate recognition.","evidence":"Kinase assays, RNAi, Aurora B inhibition, and in vitro assays with modified H3 peptides","pmids":["21658950","21397507"],"confidence":"High","gaps":["Sites on Haspin phosphorylated by Aurora B not fully mapped","In vivo relevance of methylation crosstalk not quantified"]},{"year":2012,"claim":"Inhibitor and Aurora B retargeting experiments established the epistatic Haspin→H3T3ph→CPC pathway as causal for centromeric Aurora B function and checkpoint signaling.","evidence":"Small-molecule Haspin inhibitors, anti-H3T3ph antibody injection, Aurora B retargeting bypass, and substrate immunostaining across two simultaneous papers","pmids":["23071152","23071153"],"confidence":"High","gaps":["Relative weighting versus Bub1 pathway not yet tested","Off-target inhibitor effects not fully excluded"]},{"year":2013,"claim":"Defining N-terminal autoinhibition and its relief by Cdk1-primed Plk1 phosphorylation answered how a constitutively active kinase is restricted to M phase.","evidence":"In vitro kinase assays, Xenopus egg extracts, and mutagenesis of the autoinhibitory segment and phospho-sites","pmids":["24184212"],"confidence":"High","gaps":["Precise phospho-sites controlling autoinhibition relief partially defined","Structural basis of autoinhibition not visualized"]},{"year":2014,"claim":"Plk1 binding in a Cdk1-dependent manner and the H3-tail co-crystal structure together explained activation timing and the structural basis of methylation-sensitive substrate selection, while phosphoproteomics extended substrates to macroH2A Ser137.","evidence":"Co-IP and Plk1 inhibition in cells; X-ray co-crystallography with H3 peptide, mass spectrometry, and in vitro kinase assays","pmids":["24413556","24732914"],"confidence":"High","gaps":["Functional consequence of macroH2A S137 phosphorylation inferred, not proven","macroH2A finding from single lab"]},{"year":2016,"claim":"SUMOylated Topo IIα CTD was identified as the recruiter of Haspin to centromeres, linking chromosome topology to where H3T3ph and the CPC are deposited.","evidence":"Xenopus egg extract biochemistry, SUMO-site mutagenesis, and co-immunoprecipitation","pmids":["27325792"],"confidence":"High","gaps":["Stoichiometry and dynamics of the interaction unresolved","Conservation in somatic human cells not directly shown here"]},{"year":2017,"claim":"Discovery of the Pds5B-binding YGA/R motif and direct phosphorylation of the Wapl YSR motif provided two parallel kinase-dependent and -independent mechanisms by which Haspin protects centromeric cohesion, while Aurora-A was shown to prime activation before spindle assembly.","evidence":"Co-IP, CRISPR knockout, motif and phospho-mimetic mutant rescue, in vitro kinase assays, and Aurora-A knockout/inhibition with site mapping","pmids":["28343965","29138236","28101375"],"confidence":"High","gaps":["Relative contribution of Pds5B-binding versus Wapl phosphorylation not quantified","Aurora-A pathway evidence from single lab with limited reconstitution"]},{"year":2016,"claim":"Meiotic studies in mouse oocytes showed Haspin recruits Aurora kinase C to chromosomes and acentriolar MTOCs and is needed for spindle bipolarity and accurate kinetochore-microtubule attachments, extending the pathway to a meiosis-specific Aurora.","evidence":"Haspin and AURKC inhibitors, AURKC overexpression rescue, and live imaging in oocytes","pmids":["27562071","25315835"],"confidence":"Medium","gaps":["Mechanistic basis of MTOC clustering control unresolved","Findings from single lab in mouse oocytes"]},{"year":2020,"claim":"Dual Haspin/Bub1 inhibition showed that abolishing centromeric Aurora B does not block the checkpoint or kinetochore substrate phosphorylation, refining the spatial model of how Aurora B corrects attachments, and SUMO-Topo II coupling was extended to an ectopic-site checkpoint response.","evidence":"Selective Haspin and Bub1 inhibitors, Aurora B substrate phosphorylation and segregation analysis; Topo II catalytic inhibition with SUMO-site mutants","pmids":["32027339","31712254"],"confidence":"High","gaps":["How kinetochore Aurora B activity is sustained without centromeric pool unresolved","Single-lab dual-inhibitor design"]},{"year":2021,"claim":"Interphase and non-mitotic functions emerged, with Haspin controlling primary cilia length and resorption via Dido3-HDAC6 relocalization to the basal body, broadening its role beyond chromosome segregation.","evidence":"Haspin inhibition in quiescent cells, cilia length measurement, Dido3 overexpression rescue, and zebrafish validation","pmids":["34299370"],"confidence":"Medium","gaps":["Direct substrate at the basal body not defined","Connection to H3T3ph unclear"]},{"year":2022,"claim":"A cryo-EM nucleosome complex revealed the supergroove DNA-insertion binding mode, confirmation that Haspin alone supplies mitotic H3T3ph, and identification of testis interactors/substrates expanded the structural and tissue-specific picture.","evidence":"Cryo-EM with DNA-binding mutagenesis and nucleosomal phosphorylation assays; KiPIK/CRISPR exclusion of VRK1/2; yeast two-hybrid and in vitro kinase assays from testis","pmids":["39979508","35778595","36012324"],"confidence":"High","gaps":["Functional role of C1QBP, CENPJ, KPNA6 interactions largely uncharacterized (Low confidence)","How DNA-only contacts achieve centromere targeting in vivo not fully linked"]},{"year":2023,"claim":"Linking Haspin to ARHGAP11A-Rho-ROCK-LIMK1 signaling established a cytoskeletal role controlling spindle orientation and tissue morphology, distinct from chromatin marking.","evidence":"Haspin inhibition/KO, ARHGAP11A interaction, ROCK/LIMK1 activation assays, and 3D culture morphology","pmids":["37841592"],"confidence":"Medium","gaps":["Whether Haspin acts catalytically in this axis not reconstituted","Direct substrate in the Rho pathway unidentified"]},{"year":null,"claim":"How Haspin's many spatially and temporally distinct activities—centromeric H3T3ph, cohesion protection, ciliary control, and cytoskeletal/spindle-orientation signaling—are coordinated and prioritized within a single cell remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No integrated model reconciling kinase-dependent and kinase-independent functions","Physiological substrate spectrum beyond H3T3 incompletely defined","Disease relevance not established in the available corpus"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,9,12,27]},{"term_id":"GO:0016740","term_label":"transferase activity","supporting_discovery_ids":[0,2,17]},{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[17,24]},{"term_id":"GO:0042393","term_label":"histone binding","supporting_discovery_ids":[8,24]},{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[2]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[17]},{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0,24]},{"term_id":"GO:0000228","term_label":"nuclear chromosome","supporting_discovery_ids":[4,10]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,4,15,20]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[0,8,24]},{"term_id":"R-HSA-1474165","term_label":"Reproduction","supporting_discovery_ids":[19,26]}],"complexes":[],"partners":["SURVIVIN","PDS5B","WAPL","PLK1","AURB","AURA","TOP2A","ARHGAP11A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8TF76","full_name":"Serine/threonine-protein kinase haspin","aliases":["Germ cell-specific gene 2 protein","H-haspin","Haploid germ cell-specific nuclear protein kinase"],"length_aa":798,"mass_kda":88.5,"function":"Serine/threonine-protein kinase that phosphorylates histone H3 at 'Thr-3' (H3T3ph) during mitosis. 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LIMK1-Cofilin.","date":"2023","source":"iScience","url":"https://pubmed.ncbi.nlm.nih.gov/37841592","citation_count":4,"is_preprint":false},{"pmid":"37612272","id":"PMC_37612272","title":"Haspin balances the ratio of asymmetric cell division through Wnt5a and regulates cell fate decisions in mouse embryonic stem cells.","date":"2023","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/37612272","citation_count":4,"is_preprint":false},{"pmid":"34612381","id":"PMC_34612381","title":"Exploring the thermodynamic, kinetic and inhibitory mechanisms of 5-iTU targeting mitotic kinase haspin by integrated molecular dynamics.","date":"2021","source":"Physical chemistry chemical physics : PCCP","url":"https://pubmed.ncbi.nlm.nih.gov/34612381","citation_count":4,"is_preprint":false},{"pmid":"38441546","id":"PMC_38441546","title":"GSG2 promotes thyroid cancer via stabilizing AURKB and activating AKT 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Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/35972887","citation_count":2,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":42615,"output_tokens":6653,"usd":0.11382,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":15708,"output_tokens":5743,"usd":0.111058,"stage2_stop_reason":"end_turn"},"total_usd":0.224878,"stage1_batch_id":"msgbatch_01DUPACZi7AAWwuZiUjseQF4","stage2_batch_id":"msgbatch_018yt7chE3f48PEFtDwtj6AE","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"Haspin phosphorylates histone H3 at Thr3 in vitro and is required for H3T3 phosphorylation in mitotic cells; depletion by RNAi causes metaphase chromosome misalignment and Haspin associates with chromosomes, centrosomes, and spindle during mitosis.\",\n      \"method\": \"In vitro kinase assay, RNAi depletion, immunostaining with phospho-specific antibodies\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — in vitro kinase assay establishing substrate, RNAi loss-of-function with clear cellular phenotype, replicated in subsequent studies\",\n      \"pmids\": [\"15681610\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Haspin depletion disrupts centromeric cohesin binding and sister chromatid cohesion in mitosis, causing chromosome misalignment and spindle assembly checkpoint activation; overexpression stabilizes arm cohesion, indicating Haspin is required to maintain centromeric cohesion.\",\n      \"method\": \"RNAi depletion, immunostaining for cohesin and H3T3ph, live-cell imaging\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — RNAi loss-of-function with specific cohesin phenotype, gain-of-function overexpression, replicated by subsequent studies\",\n      \"pmids\": [\"17084365\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Crystal structure of the human Haspin kinase domain reveals a constitutively active conformation stabilized by haspin-specific inserts, an atypical activation segment without phosphorylatable residues, and that His651 in the catalytic loop plays a direct role in ATP recognition (His651Ala mutant is inactive). Increasing H3K4 methylation strongly decreases substrate recognition.\",\n      \"method\": \"X-ray crystallography, active-site mutagenesis (His651Ala), enzyme kinetic assays\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — crystal structure with functional mutagenesis and enzyme kinetics, confirmed by a second independent structure paper (PMID:19918049) in the same year\",\n      \"pmids\": [\"19918057\", \"19918049\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Crystal structure of human Haspin kinase domain (2.15 Å) shows the αC helix adopts its active conformation stabilized by a helical insertion and an unprecedented activation segment conformation, explaining constitutive activity without external activation loop phosphorylation.\",\n      \"method\": \"X-ray crystallography (2.15 Å resolution)\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — high-resolution crystal structure, independently confirms findings of PMID:19918057\",\n      \"pmids\": [\"19918049\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Haspin-generated H3T3 phosphorylation provides a direct chromatin docking site for the CPC subunit Survivin at centromeres; Survivin binds directly to H3T3ph (pulldown), and a non-binding Survivin-D70A/D71A mutant fails to support centromeric CPC concentration, diminishing Aurora B centromere functions including MCAK localization and spindle checkpoint response.\",\n      \"method\": \"Direct binding assay (Survivin pulldown with H3T3ph peptide), Survivin point mutant rescue, antibody microinjection, RNAi\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — direct binding assay identifying Survivin as H3T3ph reader, mutagenesis of binding interface, replicated across multiple labs\",\n      \"pmids\": [\"20705812\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Aurora B phosphorylates Haspin to promote H3T3ph generation, establishing a positive CPC–Haspin–H3T3ph feedback loop at centromeres; Aurora B kinase activity is required for normal chromosomal CPC localization. The Bub1–shugoshin–CPC pathway provides a centromere-specific boost to this feedback loop.\",\n      \"method\": \"Kinase assays, RNAi, Aurora B inhibitor treatment, live-cell imaging\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal kinase relationship demonstrated biochemically and in cells, multiple orthogonal methods, replicated by other studies\",\n      \"pmids\": [\"21658950\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"A conserved basic segment in Haspin's N-terminus autoinhibits kinase activity during interphase; Cdk1 phosphorylation of the N-terminus recruits Plk1/Plx1, which further phosphorylates multiple N-terminal sites to neutralize autoinhibition and activate Haspin in M phase.\",\n      \"method\": \"In vitro kinase assays, Xenopus egg extract experiments, mutagenesis of autoinhibitory segment and phosphorylation sites\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Strong — biochemical reconstitution in egg extracts, mutagenesis of regulatory sites, multiple orthogonal methods\",\n      \"pmids\": [\"24184212\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Plk1 binds to Haspin in a Cdk1-phosphorylation-dependent manner and is required for initial Haspin phosphorylation and H3T3ph in prophase, positively regulating CPC recruitment at mitotic entry.\",\n      \"method\": \"Co-immunoprecipitation, Plk1 inhibitor treatment, phospho-Haspin immunostaining\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal interaction confirmed, consistent with Xenopus data from PMID:24184212, two independent studies\",\n      \"pmids\": [\"24413556\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Co-crystal structure of Haspin with the histone H3 tail reveals a unique bent substrate binding mode positioning H3 residues Arg2 and Lys4 into acidic binding pockets adjacent to the phosphorylated Thr3, explaining how H3K4 methylation reduces Haspin activity by steric/charge interference.\",\n      \"method\": \"X-ray co-crystallography, mass spectrometry-based phosphoproteomics, in vitro kinase assays\",\n      \"journal\": \"Molecular & cellular proteomics : MCP\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — co-crystal structure with substrate peptide, structural basis for methylation-dependent regulation, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"24732914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Haspin phosphorylates macroH2A at Ser137 in addition to H3T3; the Ser137 site is in a basic stretch required to stabilize extranucleosomal DNA, suggesting phosphorylation regulates macroH2A–DNA interactions.\",\n      \"method\": \"Mass spectrometry-based phosphoproteomics, in vitro kinase assays, consensus motif analysis\",\n      \"journal\": \"Molecular & cellular proteomics : MCP\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — identified by MS and validated in vitro, single lab, functional consequence inferred but not fully proven\",\n      \"pmids\": [\"24732914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"SUMOylation of DNA topoisomerase IIα CTD is required for Haspin localization at centromeres and H3T3ph; SUMOylated TOP2A CTD directly interacts with Haspin through SUMO-interaction motifs to regulate CPC recruitment in mitosis, established using Xenopus egg extracts.\",\n      \"method\": \"Xenopus egg extract biochemistry, SUMO site mutagenesis, co-immunoprecipitation\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — biochemical reconstitution in egg extracts, mutagenesis of SUMO sites, direct interaction demonstrated\",\n      \"pmids\": [\"27325792\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Haspin binds the cohesin regulatory subunit Pds5B through a conserved YGA/R motif in its non-catalytic N-terminus (similar to Wapl's YSR motif); disruption of this interaction causes weakened centromeric cohesion and premature chromatid separation, rescued by prevention of Wapl-mediated cohesin removal.\",\n      \"method\": \"Co-immunoprecipitation, CRISPR knockout, domain mutant rescue experiments\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-IP, genetic rescue with specific motif mutant, multiple orthogonal approaches\",\n      \"pmids\": [\"28343965\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"The Haspin kinase domain directly binds and phosphorylates the YSR motif of Wapl, inhibiting Wapl–Pds5B interaction and preventing Wapl-dependent cohesin release from centromeres; phospho-mimetic mutation in Wapl-YSR prevents Wapl from releasing cohesin.\",\n      \"method\": \"In vitro kinase assay, co-immunoprecipitation, phospho-mimetic mutagenesis, centromere targeting assays\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — in vitro kinase assay showing direct phosphorylation of Wapl-YSR, phospho-mimetic mutagenesis demonstrating functional consequence, complementary to PMID:28343965\",\n      \"pmids\": [\"29138236\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Aurora-A phosphorylates Haspin at multiple N-terminal sites in late G2/nuclear phase to promote H3T3ph and CPC centromere recruitment before spindle assembly, independently of Plk1; Aurora-A also facilitates association of Aurora-B with Haspin and Plk1.\",\n      \"method\": \"Aurora-A knockout/inhibition/nuclear-import blockade, phosphorylation site mapping, centromere localization assays in cells\",\n      \"journal\": \"Cell discovery\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — single lab, multiple cell-based methods but limited in vitro reconstitution\",\n      \"pmids\": [\"28101375\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"RNAi depletion of Haspin (along with Sgo1 and Scc1) causes generation of multiple acentriolar centrosome-like foci and spindle disruption only when chromatid cohesion is lost; these effects require kinesin-5 activity and microtubule dynamics, indicating spindle-pole integrity depends indirectly on chromosome cohesion.\",\n      \"method\": \"RNAi, live-cell imaging, topoisomerase II inhibition rescue, kinesin-5 inhibitor\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — live imaging with specific genetic and pharmacological perturbations, but spindle pole disruption is shown to be indirect\",\n      \"pmids\": [\"19910498\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Haspin inhibitors (and anti-H3T3ph antibody microinjection) decrease centromeric Aurora B and its substrates (CENP-A, Hec1), compromise metaphase chromosome alignment and spindle checkpoint signaling; retargeting Aurora B to centromeres bypasses the Haspin requirement, confirming the Haspin→H3T3ph→CPC centromere recruitment pathway.\",\n      \"method\": \"Small-molecule Haspin inhibitors, antibody microinjection, Aurora B retargeting, phosphosubstrate immunostaining\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple inhibitor tools plus antibody injection plus epistatic rescue (Aurora B retargeting), independently replicated in two simultaneous papers\",\n      \"pmids\": [\"23071152\", \"23071153\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Methylation at H3K4 abolishes Haspin-mediated H3T3 phosphorylation; dimethylation at H3R2 also substantially reduces Haspin activity on H3T3, establishing combinatorial histone modification control of Haspin substrate recognition.\",\n      \"method\": \"In vitro kinase assays with variously modified H3 peptides\",\n      \"journal\": \"Bioorganic & medicinal chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — in vitro kinase assays with modified peptides, single lab, consistent with structural data\",\n      \"pmids\": [\"21397507\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Haspin (mouse) has intrinsic Ser/Thr kinase activity; it localizes exclusively to nuclei; autophosphorylation regulates its DNA-binding activity (deletion mutant lacking autophosphorylation binds DNA-cellulose constitutively); ectopic expression without kinase activity causes G1 cell cycle arrest.\",\n      \"method\": \"In vitro kinase assay, nuclear localization by immunostaining, DNA-cellulose binding assay, transfection with kinase-dead mutant\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — first identification paper with in vitro kinase assay and cell-based localization, single lab\",\n      \"pmids\": [\"10358056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"In mouse oocytes, Haspin is required for Aurora kinase C (AURKC) localization at both chromosomes and acentriolar MTOCs; Haspin inhibition perturbs MTOC clustering into two poles and spindle bipolarity, and AURKC overexpression rescues these defects.\",\n      \"method\": \"Haspin inhibitor treatment, overexpression rescue, live imaging of AURKC and MTOC markers in mouse oocytes\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — inhibitor with OE rescue, multiple imaging endpoints, single lab\",\n      \"pmids\": [\"27562071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"In mouse oocytes, Haspin-mediated H3T3ph recruits the CPC along chromosome arms (rather than kinetochores), and Haspin inhibition causes improper kinetochore-microtubule attachments and aneuploidy at metaphase II; AURKC rather than AURKB mediates correction of microtubule attachments during meiosis I.\",\n      \"method\": \"Haspin inhibitor and AURKC inhibitor treatment, chromosome alignment and aneuploidy assays in mouse oocytes\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological loss-of-function with specific phenotypic readouts, single lab\",\n      \"pmids\": [\"25315835\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Either Haspin or Bub1 activity alone is sufficient to recruit Aurora B to a centromeric locus; combined inhibition of both Haspin and Bub1 fully abolishes Aurora B centromere accumulation and impairs correction of erroneous KT-MT attachments but does not compromise the mitotic checkpoint or kinetochore Aurora B substrate phosphorylation, arguing against a purely centromere-based spatial model.\",\n      \"method\": \"Selective kinase inhibitors (Haspin + Bub1), Aurora B substrate phosphorylation assays, chromosome segregation analysis\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — dual inhibitor epistasis with multiple molecular readouts, single lab but rigorous experimental design\",\n      \"pmids\": [\"32027339\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Catalytic inhibition of Topo II leads to its SUMOylation, which triggers H3T3ph via Haspin and recruitment of Aurora B to ectopic chromosomal sites; both Haspin and Aurora B are required for this metaphase checkpoint response.\",\n      \"method\": \"Topo II catalytic inhibitor, SUMOylation site mutants, Haspin and Aurora B inhibitors, centromere localization assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic (SUMOylation mutants) and pharmacological epistasis, single lab\",\n      \"pmids\": [\"31712254\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"In Drosophila, Haspin phosphorylates H3T3 both in mitosis and interphase; H3T3ph in interphase localizes to heterochromatin and lamin-enriched euchromatin; Haspin loss compromises insulator activity, reduces nuclear size, and is required for Polycomb-dependent homeotic gene silencing; Haspin associates with the cohesin complex in interphase and mediates Pds5 binding to chromatin.\",\n      \"method\": \"Haspin mutant Drosophila, ChIP, enhancer-blocking assays, live imaging, co-immunoprecipitation\",\n      \"journal\": \"PLoS genetics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic loss-of-function with multiple phenotypic and molecular readouts, single lab\",\n      \"pmids\": [\"32750047\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Haspin phosphorylates H3T3 in quiescent cells and its activity is required for primary cilia length regulation and timely cilia resorption; Haspin activity promotes relocalization of Dido3-HDAC6 to the basal body, and this pathway was confirmed in zebrafish.\",\n      \"method\": \"Haspin inhibitor in quiescent cells, cilia length measurement, Dido3 overexpression rescue, zebrafish model\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — pharmacological inhibition with specific rescue (Dido3), in vivo zebrafish validation, single lab\",\n      \"pmids\": [\"34299370\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Cryo-EM structure of human Haspin kinase domain bound to a nucleosome shows Haspin contacts only nucleosomal DNA, inserting into a supergroove formed by apposing major grooves of two DNA gyres; key basic residues essential for this DNA binding are required for phosphorylation of nucleosomal H3 and binding to mitotic chromatin.\",\n      \"method\": \"Cryo-EM structure determination, mutagenesis of DNA-binding residues, nucleosomal H3 phosphorylation assay, mitotic chromatin binding assay\",\n      \"journal\": \"Nature structural & molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — cryo-EM structure combined with mutagenesis and functional phosphorylation assay, single lab but multiple orthogonal methods\",\n      \"pmids\": [\"39979508\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"In vitro kinase assays and CRISPR/Cas9 approaches show VRK1 and VRK2 do not phosphorylate H3T3 or H3S10 in mitosis; Haspin is the kinase specifically required for H3T3ph during mitosis.\",\n      \"method\": \"In vitro kinase assays, KiPIK screening, RNAi, CRISPR/Cas9\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 / Moderate — multiple methods including in vitro assay and genetic KO, primarily a negative result for VRK1/2 that positively confirms Haspin's exclusive role\",\n      \"pmids\": [\"35778595\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Haspin phosphorylates TH2A (germ cell-specific H2A variant) at Thr127 at meiotic centromeres; identified by candidate kinase approach and confirmed in vivo using Haspin inhibitor.\",\n      \"method\": \"Haspin inhibitor in oocytes and spermatocytes, phospho-specific antibody, comparison with TH2A-T127A knock-in mice\",\n      \"journal\": \"Chromosoma\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — in vivo inhibitor confirms Haspin dependence, knock-in mouse controls, single lab\",\n      \"pmids\": [\"28803373\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Two-hybrid screening from mouse testis identified CENPJ/CPAP, KPNA6/importin alpha 6, and C1QBP/HABP1 as Haspin-interacting proteins; Haspin phosphorylates C1QBP in vitro, suggesting a role in spermatogenesis through C1QBP phosphorylation and potentially centrosome formation.\",\n      \"method\": \"Yeast two-hybrid, co-immunoprecipitation, in vitro kinase assay\",\n      \"journal\": \"International journal of molecular sciences\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — yeast two-hybrid plus in vitro kinase, single lab, limited functional validation\",\n      \"pmids\": [\"36012324\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"During mitosis, Haspin regulates Rho-ROCK activity through the GAP protein ARHGAP11A, and ROCK in turn activates LIMK1 and stabilizes the actin cytoskeleton to support spindle orientation; this pathway is required for correct tissue morphology in 3D cultures.\",\n      \"method\": \"Haspin inhibition/KO, ARHGAP11A interaction studies, ROCK/LIMK1 activation assays, 3D cell culture morphology\",\n      \"journal\": \"iScience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — epistasis established in cells with multiple molecular readouts, single lab, limited biochemical reconstitution\",\n      \"pmids\": [\"37841592\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"Haspin is an atypical, constitutively active Ser/Thr kinase that phosphorylates histone H3 at Thr3 (H3T3ph) during mitosis by binding nucleosomal DNA via a supergroove-insertion mode; this mark is read by the CPC subunit Survivin to recruit Aurora B to inner centromeres, where it regulates kinetochore-microtubule attachments and spindle checkpoint signaling. Haspin activity is cell-cycle controlled through Cdk1-dependent autoinhibition neutralization followed by Plk1- and Aurora A-mediated multi-site N-terminal phosphorylation; Aurora B in turn phosphorylates Haspin in a positive feedback loop. Beyond Aurora B positioning, Haspin protects centromeric cohesion by (1) binding cohesin subunit Pds5B via a YGA/R motif to compete with Wapl, and (2) directly phosphorylating Wapl's YSR motif to prevent Wapl–Pds5B interaction. Haspin also phosphorylates macroH2A at Ser137, TH2A at Thr127 in germ cells, and C1QBP in spermatids, and regulates primary cilia dynamics and spindle orientation through ARHGAP11A-Rho-ROCK-LIMK1 in interphase cells.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"Haspin is an atypical Ser/Thr kinase that governs the spatial organization of mitotic chromatin signaling, principally by phosphorylating histone H3 at Thr3 (H3T3ph) during mitosis [#0]. Crystal structures show that Haspin is constitutively active: haspin-specific helical inserts and an unprecedented activation-segment conformation hold the αC helix and catalytic apparatus in an active state without canonical activation-loop phosphorylation, with His651 directly mediating ATP recognition [#2, #3]. Substrate engagement is highly selective—co-crystal and peptide studies reveal a bent H3-tail binding mode that positions Arg2 and Lys4 in acidic pockets, so that H3K4 methylation and H3R2 dimethylation sterically and electrostatically suppress phosphorylation [#8, #16]—and a cryo-EM nucleosome complex shows Haspin contacts only nucleosomal DNA by inserting into a supergroove between two DNA gyres, a mode required for phosphorylation of chromatin-embedded H3 [#24]. The H3T3ph mark is read directly by the chromosomal passenger complex (CPC) subunit Survivin, concentrating Aurora B at inner centromeres to control MCAK localization, kinetochore-microtubule attachments, and the spindle checkpoint [#4, #15]. Haspin activity is cell-cycle gated: an autoinhibitory N-terminal basic segment is relieved when Cdk1 phosphorylation recruits Plk1 for multi-site activating phosphorylation at mitotic entry, while Aurora-A primes the pathway in late G2 and Aurora B feeds back positively onto Haspin [#5, #6, #7, #13]. In parallel with Aurora B positioning, Haspin protects centromeric cohesion both by binding the cohesin regulator Pds5B through a conserved YGA/R motif and by directly phosphorylating the Wapl YSR motif to block Wapl-mediated cohesin release [#1, #11, #12]. Haspin centromere recruitment depends on SUMOylated topoisomerase IIα, linking Topo II catalytic state to H3T3ph-based checkpoint responses [#10, #21]. Beyond mitosis, Haspin functions in meiosis to recruit Aurora kinase C in oocytes [#18, #19], in primary cilia length and resorption [#23], and in spindle orientation through an ARHGAP11A-Rho-ROCK-LIMK1 axis [#28].\",\n  \"teleology\": [\n    {\n      \"year\": 1999,\n      \"claim\": \"Establishing that Haspin was a genuine nuclear Ser/Thr kinase whose own autophosphorylation modulated DNA binding set the foundation for treating it as a chromatin-associated enzyme rather than an orphan sequence.\",\n      \"evidence\": \"In vitro kinase assay, nuclear immunostaining, DNA-cellulose binding, and kinase-dead transfection in mouse cells\",\n      \"pmids\": [\"10358056\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No physiological substrate identified at this stage\", \"Mechanism linking autophosphorylation to DNA binding not structurally resolved\", \"Cell-cycle regulation of activity unaddressed\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Identifying H3 Thr3 as Haspin's mitotic substrate answered what the kinase does on chromatin and tied it to chromosome alignment.\",\n      \"evidence\": \"In vitro kinase assay, RNAi depletion, and phospho-specific immunostaining in mitotic cells\",\n      \"pmids\": [\"15681610\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Downstream reader of H3T3ph unknown\", \"Structural basis of substrate specificity unresolved\", \"How misalignment arises mechanistically not defined\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Demonstrating that Haspin maintains centromeric cohesin and sister chromatid cohesion revealed a second, distinct mitotic role beyond histone marking.\",\n      \"evidence\": \"RNAi depletion, cohesin immunostaining, overexpression, and live-cell imaging\",\n      \"pmids\": [\"17084365\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Molecular link between Haspin and cohesin retention not defined\", \"Whether this required kinase activity or H3T3ph unresolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Crystal structures explained how Haspin is constitutively active without activation-loop phosphorylation, answering why an atypical kinase still adopts a productive conformation.\",\n      \"evidence\": \"X-ray crystallography of the kinase domain plus His651Ala mutagenesis and enzyme kinetics, with an independent confirmatory structure\",\n      \"pmids\": [\"19918057\", \"19918049\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How cell-cycle regulation overrides constitutive activity unexplained\", \"Nucleosomal substrate engagement not captured\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identifying Survivin as a direct reader of H3T3ph closed the loop between Haspin's mark and CPC/Aurora B centromere recruitment.\",\n      \"evidence\": \"Survivin pulldown with H3T3ph peptide, non-binding D70A/D71A mutant rescue, antibody microinjection, and RNAi\",\n      \"pmids\": [\"20705812\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Quantitative contribution relative to other CPC targeting pathways unresolved\", \"Spatial dynamics of recruitment not addressed\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showing Aurora B phosphorylates Haspin and that H3K4/H3R2 methylation blocks H3T3ph defined both a positive feedback loop and combinatorial histone control of substrate recognition.\",\n      \"evidence\": \"Kinase assays, RNAi, Aurora B inhibition, and in vitro assays with modified H3 peptides\",\n      \"pmids\": [\"21658950\", \"21397507\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Sites on Haspin phosphorylated by Aurora B not fully mapped\", \"In vivo relevance of methylation crosstalk not quantified\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Inhibitor and Aurora B retargeting experiments established the epistatic Haspin→H3T3ph→CPC pathway as causal for centromeric Aurora B function and checkpoint signaling.\",\n      \"evidence\": \"Small-molecule Haspin inhibitors, anti-H3T3ph antibody injection, Aurora B retargeting bypass, and substrate immunostaining across two simultaneous papers\",\n      \"pmids\": [\"23071152\", \"23071153\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Relative weighting versus Bub1 pathway not yet tested\", \"Off-target inhibitor effects not fully excluded\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Defining N-terminal autoinhibition and its relief by Cdk1-primed Plk1 phosphorylation answered how a constitutively active kinase is restricted to M phase.\",\n      \"evidence\": \"In vitro kinase assays, Xenopus egg extracts, and mutagenesis of the autoinhibitory segment and phospho-sites\",\n      \"pmids\": [\"24184212\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Precise phospho-sites controlling autoinhibition relief partially defined\", \"Structural basis of autoinhibition not visualized\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Plk1 binding in a Cdk1-dependent manner and the H3-tail co-crystal structure together explained activation timing and the structural basis of methylation-sensitive substrate selection, while phosphoproteomics extended substrates to macroH2A Ser137.\",\n      \"evidence\": \"Co-IP and Plk1 inhibition in cells; X-ray co-crystallography with H3 peptide, mass spectrometry, and in vitro kinase assays\",\n      \"pmids\": [\"24413556\", \"24732914\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Functional consequence of macroH2A S137 phosphorylation inferred, not proven\", \"macroH2A finding from single lab\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"SUMOylated Topo IIα CTD was identified as the recruiter of Haspin to centromeres, linking chromosome topology to where H3T3ph and the CPC are deposited.\",\n      \"evidence\": \"Xenopus egg extract biochemistry, SUMO-site mutagenesis, and co-immunoprecipitation\",\n      \"pmids\": [\"27325792\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Stoichiometry and dynamics of the interaction unresolved\", \"Conservation in somatic human cells not directly shown here\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Discovery of the Pds5B-binding YGA/R motif and direct phosphorylation of the Wapl YSR motif provided two parallel kinase-dependent and -independent mechanisms by which Haspin protects centromeric cohesion, while Aurora-A was shown to prime activation before spindle assembly.\",\n      \"evidence\": \"Co-IP, CRISPR knockout, motif and phospho-mimetic mutant rescue, in vitro kinase assays, and Aurora-A knockout/inhibition with site mapping\",\n      \"pmids\": [\"28343965\", \"29138236\", \"28101375\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Relative contribution of Pds5B-binding versus Wapl phosphorylation not quantified\", \"Aurora-A pathway evidence from single lab with limited reconstitution\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Meiotic studies in mouse oocytes showed Haspin recruits Aurora kinase C to chromosomes and acentriolar MTOCs and is needed for spindle bipolarity and accurate kinetochore-microtubule attachments, extending the pathway to a meiosis-specific Aurora.\",\n      \"evidence\": \"Haspin and AURKC inhibitors, AURKC overexpression rescue, and live imaging in oocytes\",\n      \"pmids\": [\"27562071\", \"25315835\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Mechanistic basis of MTOC clustering control unresolved\", \"Findings from single lab in mouse oocytes\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Dual Haspin/Bub1 inhibition showed that abolishing centromeric Aurora B does not block the checkpoint or kinetochore substrate phosphorylation, refining the spatial model of how Aurora B corrects attachments, and SUMO-Topo II coupling was extended to an ectopic-site checkpoint response.\",\n      \"evidence\": \"Selective Haspin and Bub1 inhibitors, Aurora B substrate phosphorylation and segregation analysis; Topo II catalytic inhibition with SUMO-site mutants\",\n      \"pmids\": [\"32027339\", \"31712254\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"How kinetochore Aurora B activity is sustained without centromeric pool unresolved\", \"Single-lab dual-inhibitor design\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Interphase and non-mitotic functions emerged, with Haspin controlling primary cilia length and resorption via Dido3-HDAC6 relocalization to the basal body, broadening its role beyond chromosome segregation.\",\n      \"evidence\": \"Haspin inhibition in quiescent cells, cilia length measurement, Dido3 overexpression rescue, and zebrafish validation\",\n      \"pmids\": [\"34299370\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Direct substrate at the basal body not defined\", \"Connection to H3T3ph unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"A cryo-EM nucleosome complex revealed the supergroove DNA-insertion binding mode, confirmation that Haspin alone supplies mitotic H3T3ph, and identification of testis interactors/substrates expanded the structural and tissue-specific picture.\",\n      \"evidence\": \"Cryo-EM with DNA-binding mutagenesis and nucleosomal phosphorylation assays; KiPIK/CRISPR exclusion of VRK1/2; yeast two-hybrid and in vitro kinase assays from testis\",\n      \"pmids\": [\"39979508\", \"35778595\", \"36012324\"],\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Functional role of C1QBP, CENPJ, KPNA6 interactions largely uncharacterized (Low confidence)\", \"How DNA-only contacts achieve centromere targeting in vivo not fully linked\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Linking Haspin to ARHGAP11A-Rho-ROCK-LIMK1 signaling established a cytoskeletal role controlling spindle orientation and tissue morphology, distinct from chromatin marking.\",\n      \"evidence\": \"Haspin inhibition/KO, ARHGAP11A interaction, ROCK/LIMK1 activation assays, and 3D culture morphology\",\n      \"pmids\": [\"37841592\"],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"Whether Haspin acts catalytically in this axis not reconstituted\", \"Direct substrate in the Rho pathway unidentified\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How Haspin's many spatially and temporally distinct activities—centromeric H3T3ph, cohesion protection, ciliary control, and cytoskeletal/spindle-orientation signaling—are coordinated and prioritized within a single cell remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"\",\n      \"gaps\": [\"No integrated model reconciling kinase-dependent and kinase-independent functions\", \"Physiological substrate spectrum beyond H3T3 incompletely defined\", \"Disease relevance not established in the available corpus\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 9, 12, 27]},\n      {\"term_id\": \"GO:0016740\", \"supporting_discovery_ids\": [0, 2, 17]},\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [17, 24]},\n      {\"term_id\": \"GO:0042393\", \"supporting_discovery_ids\": [8, 24]},\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [17]},\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0, 24]},\n      {\"term_id\": \"GO:0000228\", \"supporting_discovery_ids\": [4, 10]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 4, 15, 20]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [0, 8, 24]},\n      {\"term_id\": \"R-HSA-1474165\", \"supporting_discovery_ids\": [19, 26]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"Survivin\", \"Pds5B\", \"Wapl\", \"Plk1\", \"AurB\", \"AurA\", \"TOP2A\", \"ARHGAP11A\"],\n    \"other_free_text\": []\n  }\n}\n```","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":8,"faith_total":8,"faith_pct":100.0}}