{"gene":"KIF11","run_date":"2026-04-28T18:30:27","timeline":{"discoveries":[{"year":1995,"finding":"Eg5 (KIF11) spindle localization is cell-cycle-regulated and requires phosphorylation at a conserved threonine (T937) in the C-terminal domain; mutation of T937 to non-phosphorylatable alanine abolishes spindle localization, while T937S preserves it, implicating p34cdc2 phosphorylation as the regulatory mechanism.","method":"Transient transfection of myc-tagged Eg5 deletion/point mutants in Xenopus A6 cells; immunofluorescence localization","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — site-directed mutagenesis of candidate phosphorylation site with clear localization phenotype, replicated with multiple mutant constructs","pmids":["7753799"],"is_preprint":false},{"year":1994,"finding":"Eg5 protein associates with both spindle and interphase microtubules during Xenopus early development, concentrating toward spindle poles during meiosis and mitosis; copellets with microtubules from egg homogenates, demonstrating direct microtubule association.","method":"Immunofluorescence and microtubule co-pelleting from Xenopus egg extracts; Western blotting across developmental stages","journal":"Developmental biology","confidence":"High","confidence_rationale":"Tier 2 — biochemical co-sedimentation plus immunofluorescence in multiple developmental contexts","pmids":["8026619"],"is_preprint":false},{"year":1998,"finding":"Eg5 (KIF11) continues to be expressed in postmitotic rodent neurons, where it associates directly with the microtubule array and is enriched in distal regions of developing axons and dendrites, suggesting a role in regulating microtubule organization in neuronal processes.","method":"Immunofluorescence and subcellular fractionation in rodent neurons; expression profiling across developmental stages","journal":"The Journal of neuroscience","confidence":"Medium","confidence_rationale":"Tier 2 — direct localization by immunofluorescence in postmitotic neurons with functional implication, single lab","pmids":["9742151"],"is_preprint":false},{"year":1999,"finding":"HSET (minus-end motor) and Eg5 (KIF11, plus-end motor) act antagonistically in spindle assembly: simultaneous inhibition of both motors in cells restores centrosome separation and bipolar spindle formation that is blocked by Eg5 inhibition alone, establishing a balance-of-forces model for spindle bipolarity.","method":"Antibody microinjection in murine oocytes and cultured cells; immunoEM localization; in vitro aster assembly assays","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 — reconstitution/in vitro assays combined with microinjection epistasis, replicated across multiple cell systems","pmids":["10525540"],"is_preprint":false},{"year":2004,"finding":"Monomeric human Eg5 is a slow plus-end-directed ATPase motor; kinetic analysis shows mantATP binds at 2–3 µM⁻¹s⁻¹, ATP hydrolysis occurs at 15 s⁻¹, and ATP-dependent microtubule dissociation is rate-limiting at ~8 s⁻¹, with mantADP release at 40 s⁻¹, distinguishing Eg5 mechanism from minus-end motors where ADP release is rate-limiting.","method":"In vitro ATPase kinetics (steady-state and transient), analytical ultracentrifugation, gel filtration, fluorescent nucleotide (mantATP/mantADP) binding assays with purified Eg5-367 and Eg5-437 constructs","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — rigorous in vitro reconstitution with multiple orthogonal kinetic methods on purified protein","pmids":["15247293"],"is_preprint":false},{"year":2008,"finding":"TPX2 interacts with Eg5 via its C-terminal domain; this interaction is essential for Eg5-dependent spindle pole segregation, as TPX2-CT injection arrests spindle poles and Eg5 co-injection rescues cleavage, defining a discrete Eg5-interacting domain in TPX2.","method":"Blastomere microinjection in Xenopus embryos; cell transfection; co-immunoprecipitation; deletion analysis of TPX2","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 1–2 — domain mapping combined with functional rescue experiments in two independent systems","pmids":["18372177"],"is_preprint":false},{"year":2008,"finding":"Parkin (E3 ubiquitin ligase) represses Eg5 transcription by causing multiple monoubiquitination of Hsp70, which inactivates JNK, reducing c-Jun phosphorylation and thereby blocking c-Jun binding to the AP-1 site in the Eg5 promoter; this is not via proteasomal Eg5 degradation.","method":"Co-immunoprecipitation, ubiquitination assays, Eg5 promoter-reporter assay, ChIP, siRNA knockdown, Western blotting","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods (ChIP, reporter assay, ubiquitination assay) establishing transcriptional regulatory mechanism","pmids":["18845538"],"is_preprint":false},{"year":2009,"finding":"Kif15 (kinesin-12) cooperates with Eg5 to promote bipolar spindle assembly; Kif15 can fully replace Eg5 for centrosome separation when overexpressed, and this activity requires both its motor domain and interaction with the microtubule-associated protein TPX2, suggesting a Kif15–TPX2 complex slides antiparallel microtubules apart.","method":"siRNA depletion, ectopic overexpression, live-cell imaging, co-immunoprecipitation in human cell lines","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 — epistasis plus interaction mapping, multiple orthogonal methods, replicated across conditions","pmids":["19818618"],"is_preprint":false},{"year":2010,"finding":"Tiam1–Rac1 signaling at centrosomes during prophase functions as the first identified antagonist of Eg5-driven centrosome separation; Tiam1/Rac1 depletion allows cells to escape Eg5-inhibitor-induced mitotic arrest, and re-inhibition of Eg5 in Tiam1-depleted cells rescues chromosome congression errors, placing Tiam1–Rac upstream of Eg5 forces in bipolar spindle assembly.","method":"siRNA depletion in culture cells, live-cell imaging, Rac1-deficient epithelial cells in vivo, epistasis double-inhibition experiments","journal":"Current biology : CB","confidence":"High","confidence_rationale":"Tier 2 — genetic epistasis in vitro and in vivo with multiple orthogonal approaches","pmids":["20346677"],"is_preprint":false},{"year":2011,"finding":"CDK1 phosphorylates Eg5 at Thr926 (T927), and Plk1 controls centrosome separation additionally through activation of the Nek9–Nek6/7 kinase cascade, which phosphorylates Eg5 at Ser1033; both CDK1 (Thr926) and Nek6/7 (Ser1033) phosphorylation sites contribute to Eg5 accumulation at centrosomes and are necessary for centrosome separation.","method":"In vitro kinase assays, phospho-specific antibodies, siRNA/inhibitor epistasis, immunofluorescence in human cell lines","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro kinase assays plus cellular epistasis, two independent sites characterized","pmids":["21642957"],"is_preprint":false},{"year":2011,"finding":"Plk1 controls Eg5 localization to the centrosome in G2 phase and triggers centrosome separation independently of CDK1; CDK1 promotes fast centrosome separation while Plk1-driven separation is slow and opposed by actin-dependent forces; Cdk2 can also phosphorylate Eg5 at Thr927.","method":"Plk1/CDK1/CDK2 inhibitors, RNAi, phospho-specific antibodies, live-cell imaging, pharmacological manipulation of microtubule and actin dynamics","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 2 — multiple kinase inhibitors/knockdowns with live imaging and phospho-antibody validation","pmids":["21522128"],"is_preprint":false},{"year":2012,"finding":"Dynein does not directly antagonize Eg5 via a simple push-pull mechanism at the spindle equator; instead, live-cell imaging shows dynein and Eg5 activities cannot be titrated against each other, indicating indirect antagonism at different spindle locations.","method":"Live-cell imaging, endpoint quantification controls, mathematical modeling, siRNA depletion of dynein/Lis1","journal":"Cell reports","confidence":"Medium","confidence_rationale":"Tier 2 — live imaging plus mathematical modeling, single study","pmids":["22832270"],"is_preprint":false},{"year":2014,"finding":"Kinetochore-microtubule stability governs the requirement for Eg5 in metaphase bipolar spindle maintenance; cells with more stable K-fibers can maintain bipolar spindles without Eg5, while artificial destabilization of K-MTs promotes spindle collapse without Eg5, and stabilization rescues bipolarity.","method":"Sibling cell line comparison, pharmacological K-MT stabilization/destabilization, Eg5 inhibitor treatment, immunofluorescence quantification","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 — pharmacological epistasis with quantitative phenotypic readouts, single lab","pmids":["24807901"],"is_preprint":false},{"year":2015,"finding":"KIF11 physically interacts with ZBP1 (IGF2BP1) and is a component of β-actin mRNP complexes; KIF11 colocalizes with β-actin mRNA and its ability to transport β-actin mRNA is dependent on ZBP1; disruption of the KIF11–ZBP1 interaction in vivo delocalizes β-actin mRNA and impairs cell migration.","method":"Co-immunoprecipitation, RNA immunoprecipitation, in vitro binding assay with domain mapping, smFISH colocalization, shRNA knockdown with rescue, migration assay","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-IP, domain mapping, functional rescue, multiple orthogonal methods","pmids":["25588836"],"is_preprint":false},{"year":2015,"finding":"TPX2 inhibits Eg5 motor activity through two mechanisms: direct interaction with Eg5 (requiring dimerization or the neck region) and binding to microtubules; full-length TPX2 dramatically reduces Eg5 velocity at single-molecule level, while truncated TPX2 lacking the Eg5-interaction domain is less inhibitory.","method":"TIRF single-molecule motility assays with mammalian cell extracts expressing Eg5-EGFP; microtubule gliding assays; truncation analysis of TPX2","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 1 — single-molecule reconstitution with domain mutants, two orthogonal in vitro assays","pmids":["26018074"],"is_preprint":false},{"year":2016,"finding":"The RNF20/40 ubiquitin ligase complex interacts with Eg5 during mitosis and monoubiquitinates Eg5, stabilizing it; loss of RNF20/40 causes spindle assembly defects, cell cycle arrest and apoptosis, and depletes Eg5 protein levels.","method":"Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, Western blotting, spindle phenotype analysis","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro ubiquitination reconstitution plus cellular co-IP and functional phenotype, multiple methods","pmids":["27557628"],"is_preprint":false},{"year":2016,"finding":"PTEN associates and co-localizes with EG5 during mitosis; PTEN deficiency causes aberrant EG5 phosphorylation and abrogates EG5 recruitment to the mitotic spindle, leading to shorter spindles and chromosome misalignment.","method":"Co-immunoprecipitation, immunofluorescence co-localization, PTEN siRNA knockdown, phospho-EG5 Western blotting","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-IP, functional KD phenotype, phospho-specific antibodies, multiple methods","pmids":["27492783"],"is_preprint":false},{"year":2017,"finding":"HDAC1 deacetylates Eg5 (KIF11); HDAC1 co-localizes with Eg5 during mitosis and influences its ATPase activity; HDAC1/2-selective inhibition causes mitotic arrest and monopolar spindle formation consistent with Eg5 acetylation impairing function, identifying Eg5 as a non-histone HDAC1 substrate regulating mitotic progression.","method":"HDAC1 trapping mutant strategy to identify substrates, co-localization by immunofluorescence, ATPase activity assay, HDAC inhibitor treatment with spindle phenotype analysis","journal":"Cell chemical biology","confidence":"High","confidence_rationale":"Tier 1–2 — substrate trapping, enzymatic assay, and functional cellular phenotype in one study","pmids":["28392145"],"is_preprint":false},{"year":2017,"finding":"PP2A/B55α complex physically associates with the EG5 C-terminal tail domain and dephosphorylates EG5 at Thr926, enabling mitotic exit; PP2A knockdown leads to elevated phospho-EG5 in late metaphase and delayed mitotic exit, similar to EG5/T926D phosphomimetic mutant.","method":"Co-immunoprecipitation, in vitro phosphatase assay, phospho-specific antibodies, siRNA knockdown, EG5 phosphomimetic mutant transfection, live-cell imaging","journal":"Scientific reports","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro phosphatase assay plus cellular genetics with phosphomimetic validation","pmids":["28487562"],"is_preprint":false},{"year":2018,"finding":"NEK7 phosphorylates Eg5/KIF11, promoting its accumulation on microtubules in distal dendrites; in this postmitotic context, Eg5 limits retrograde microtubule polymerization independently of its motor activity, thereby promoting dendrite growth and branching.","method":"In vitro kinase assay, RNAi knockdown in neurons (in vitro and in vivo), NEK7 kinase-dead mutant, pharmacological Eg5 inhibition, live imaging of microtubule dynamics","journal":"Nature communications","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro kinase assay plus neuronal KD/KO with functional phenotype, in vivo validation","pmids":["29899413"],"is_preprint":false},{"year":2018,"finding":"HMMR dampens Eg5-mediated forces by localizing TPX2 and promoting formation of inhibitory TPX2–Eg5 complexes; HMMR silencing disturbs spindle microtubule organization and causes elevated aneuploidy, and Eg5 chemical inhibition rescues spindle defects in HMMR-depleted cells.","method":"siRNA silencing, co-immunoprecipitation, Eg5 inhibitor rescue, immunofluorescence, CRISPR deletion","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 2 — co-IP combined with pharmacological rescue epistasis and CRISPR validation","pmids":["29386294"],"is_preprint":false},{"year":2014,"finding":"HIV-1 Tat protein interacts with Eg5 and allosterically modulates its ATPase activity by affecting ADP release; lysine 85 in the Tat C-terminus is critical for this interaction and for Tat-induced mitotic arrest and apoptosis in CD4+ T lymphocytes.","method":"Co-immunoprecipitation, ATPase activity assay (ADP release kinetics), site-directed mutagenesis of Tat K85, siRNA, cell cycle/apoptosis analysis","journal":"The Journal of pathology","confidence":"High","confidence_rationale":"Tier 1–2 — in vitro ATPase assay with mutagenesis plus cellular functional phenotype","pmids":["24488929"],"is_preprint":false},{"year":2021,"finding":"EG5/kinesin-5 together with PRC1-dependent KIF4A/kinesin-4 constitute two mechanistically distinct microtubule-sliding modules that drive spindle elongation in human cells; disruption of both EG5 and KIF4A blocks midzone microtubule sliding and causes total failure of chromosome segregation despite poleward chromosome motion.","method":"Combined siRNA depletion and pharmacological inactivation, CRISPR, tubulin photoactivation, STED microscopy, expansion microscopy","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 1–2 — multiple redundancy-testing approaches including CRISPR plus live imaging and super-resolution, rigorous controls","pmids":["33910056"],"is_preprint":false},{"year":2018,"finding":"KIF11 knockdown in hippocampal neurons increases excitatory post-synaptic current frequency and amplitude, enhances dendritic arborization and synapse number, and requires pre-synaptic NMDAR activity for its effect; expression of Piccolo (active zone protein) constrains KIF11 function in synaptic transmission.","method":"RNAi-mediated knockdown, electrophysiological recordings (EPSCs) in hippocampal neurons, immunofluorescence morphometry, NMDAR pharmacological blockade","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — electrophysiology plus KD with pharmacological pathway dissection, single lab","pmids":["30479371"],"is_preprint":false},{"year":2007,"finding":"Homozygous knockout of Eg5 (Knsl1) causes early embryonic lethality prior to implantation in mice, with reduced cell numbers and failure of compaction; heterozygous mice are normal and fertile, establishing that Eg5 is essential for early mammalian cell division.","method":"Gene targeting (knockout mouse), embryo recovery at E2.5–3.5, morphological analysis","journal":"Biochemical and biophysical research communications","confidence":"High","confidence_rationale":"Tier 2 — clean genetic knockout with defined developmental phenotype, confirmed by independent study (PMID 18474226)","pmids":["17449012","18474226"],"is_preprint":false},{"year":2016,"finding":"Aurora A promotes bipolar spindle assembly in Eg5-independent cells by phosphorylating Kif15, enhancing Kif15 localization to the spindle; MCAK and Kif18b promote bipolar spindle assembly by destabilizing astral microtubules, placing Aurora A and Kif15 in a parallel pathway to Eg5 for centrosome separation.","method":"Genome-wide siRNA screen in Eg5-independent cells, Aurora A kinase assays, live-cell imaging, phospho-Kif15 antibodies","journal":"Chromosoma","confidence":"High","confidence_rationale":"Tier 2 — genome-wide screen validated with kinase assay and live imaging, mechanistic pathway placement","pmids":["27354041"],"is_preprint":false},{"year":2020,"finding":"Postnatal endothelial cell-specific conditional knockout of Kif11 in mice leads to severely stunted retinal vasculature growth and mildly stunted cerebellar vasculature, demonstrating that Kif11 function in mitotically active endothelial cells is most critical in rapidly growing CNS regions and that retinal hypovascularization can arise from impaired mitotic spindle function independently of Wnt/β-catenin signaling.","method":"Conditional knockout mouse (Cre-lox), retinal vascular whole-mount imaging, β-catenin signaling analysis","journal":"Human molecular genetics","confidence":"High","confidence_rationale":"Tier 2 — clean conditional KO in defined cell type with mechanistic pathway exclusion","pmids":["31993640"],"is_preprint":false},{"year":2023,"finding":"NEAT1 lncRNA, when it translocates to the cytosol during senescence, interacts with KIF11 motor protein and promotes KIF11 protein degradation; KIF11 degradation leads to increased CDKN2A (p16/p14ARF) expression, inducing cellular senescence in hepatoma cells.","method":"RNA pulldown, mass spectrometry, co-immunoprecipitation, RNA FISH/immunofluorescence co-localization, ChIP, luciferase reporter assay, siRNA/shRNA knockdown, mouse xenograft model","journal":"Clinical and translational medicine","confidence":"High","confidence_rationale":"Tier 1–2 — RNA pulldown + MS identification + co-IP validation + functional rescue with multiple orthogonal methods","pmids":["37752791"],"is_preprint":false},{"year":2020,"finding":"PAK6 specifically co-localizes with Eg5 at the centrosome; PAK6 depletion upregulates Eg5 and causes multipolar spindles, while simultaneous Eg5 knockdown rescues this phenotype, establishing that PAK6 and Eg5 negatively inter-regulate each other to control spindle formation.","method":"Co-localization by immunofluorescence, siRNA knockdown, genetic epistasis (double knockdown rescue), Western blotting","journal":"Biochimica et biophysica acta. Molecular cell research","confidence":"Medium","confidence_rationale":"Tier 2 — co-localization and epistasis, single lab","pmids":["33098954"],"is_preprint":false},{"year":2018,"finding":"Death receptor 6 (DR6) physically interacts with KIF11, as confirmed by mass spectrometry co-purification, co-immunoprecipitation, and GST pull-down; overexpression of KIF11 eliminates suppression of ovarian cancer cell migration by DR6 knockdown, placing KIF11 downstream of DR6 in a pro-migratory signaling complex.","method":"Mass spectrometry, co-immunoprecipitation, GST pull-down, overexpression rescue assay, migration assay","journal":"FEBS open bio","confidence":"Medium","confidence_rationale":"Tier 2–3 — MS identification confirmed by co-IP and pulldown plus functional rescue, single lab","pmids":["30186750"],"is_preprint":false}],"current_model":"KIF11/Eg5 is a homotetrameric, plus-end-directed kinesin-5 motor that crosslinks and slides antiparallel microtubules to drive centrosome separation and bipolar mitotic spindle assembly; its activity is tightly regulated by CDK1-mediated phosphorylation at Thr926/927 (which promotes spindle recruitment), Plk1/Nek9/Nek6-7 phosphorylation at Ser1033 (centrosome targeting), PP2A/B55α dephosphorylation (mitotic exit), HDAC1 deacetylation (ATPase regulation), and RNF20/40-dependent monoubiquitination (protein stabilization); it is opposed by HSET/minus-end motors, inhibited by TPX2 binding, and balanced by Kif15–TPX2 and other redundant sliding modules during spindle elongation; beyond mitosis, KIF11 mediates ZBP1-dependent β-actin mRNA transport to regulate cell polarity and migration, promotes dendrite morphogenesis via NEK7 phosphorylation, and functions as an inhibitory constraint on synaptic transmission in neurons."},"narrative":{"teleology":[{"year":1994,"claim":"Establishing that Eg5 is a bona fide microtubule-associated motor present on spindles and interphase microtubules resolved the question of whether kinesin-5 family members physically engage the mitotic apparatus.","evidence":"Immunofluorescence and microtubule co-pelleting from Xenopus egg extracts across developmental stages","pmids":["8026619"],"confidence":"High","gaps":["Motor directionality and enzymatic mechanism not yet characterized","No functional loss-of-function data"]},{"year":1995,"claim":"Demonstrating that p34cdc2 (CDK1) phosphorylation of Thr937 is required for Eg5 spindle localization established the first regulatory switch controlling motor targeting to the mitotic apparatus.","evidence":"Site-directed mutagenesis of T937A/T937S in Xenopus A6 cells with immunofluorescence localization","pmids":["7753799"],"confidence":"High","gaps":["Whether additional kinases contribute to centrosome-specific targeting","Phosphatase responsible for dephosphorylation unknown"]},{"year":1999,"claim":"Showing that simultaneous inhibition of HSET and Eg5 restores bipolarity established the balance-of-forces model, answering how opposing motors maintain spindle architecture.","evidence":"Antibody microinjection epistasis in murine oocytes and cultured cells combined with in vitro aster assembly","pmids":["10525540"],"confidence":"High","gaps":["Whether dynein contributes as a direct Eg5 antagonist","Quantitative force contributions of each motor unknown"]},{"year":2004,"claim":"Kinetic dissection of the Eg5 ATPase cycle revealed that ATP-dependent microtubule dissociation is rate-limiting, distinguishing Eg5 from conventional kinesin and explaining its slow processivity.","evidence":"Transient-state fluorescent nucleotide kinetics with purified monomeric Eg5 constructs","pmids":["15247293"],"confidence":"High","gaps":["How tetramerization alters the kinetic cycle","No structural explanation for the rate-limiting step"]},{"year":2007,"claim":"Homozygous Eg5 knockout causing pre-implantation lethality in mice definitively proved that Eg5 is essential for early mammalian cell division and cannot be compensated by other motors at this stage.","evidence":"Gene targeting (knockout mouse) with embryo recovery at E2.5–3.5","pmids":["17449012","18474226"],"confidence":"High","gaps":["Cell-type-specific requirements in later development unexplored","Whether Kif15 can partially rescue in specific lineages"]},{"year":2008,"claim":"Identification of the TPX2 C-terminal domain as an Eg5-interacting element required for spindle pole separation, and of Parkin-mediated transcriptional repression of Eg5 via Hsp70 ubiquitination, defined two new layers of Eg5 regulation — one physical, one transcriptional.","evidence":"TPX2: co-IP and domain deletion with blastomere rescue in Xenopus; Parkin: ChIP, promoter-reporter, ubiquitination assays","pmids":["18372177","18845538"],"confidence":"High","gaps":["Whether TPX2 inhibits Eg5 enzymatic activity directly (resolved later)","Physiological contexts where Parkin regulation of Eg5 is relevant in vivo"]},{"year":2009,"claim":"Discovery that Kif15 can fully substitute for Eg5 in centrosome separation when overexpressed revealed a redundant TPX2-dependent sliding pathway, explaining how some cancer cells escape Eg5 inhibitors.","evidence":"siRNA depletion and ectopic overexpression with live-cell imaging and co-IP in human cells","pmids":["19818618"],"confidence":"High","gaps":["How Kif15 is regulated to become Eg5-independent","Whether endogenous Kif15 levels ever suffice without overexpression"]},{"year":2011,"claim":"Mapping dual phosphorylation inputs — CDK1 at Thr926 for spindle recruitment and Plk1→Nek9→Nek6/7 at Ser1033 for centrosome targeting — resolved how two independent kinase cascades converge to control Eg5 localization at distinct mitotic stages.","evidence":"In vitro kinase assays, phospho-specific antibodies, and siRNA/inhibitor epistasis in human cell lines","pmids":["21642957","21522128"],"confidence":"High","gaps":["Whether other phosphorylation sites contribute","How actin-dependent forces oppose Plk1-driven slow separation"]},{"year":2015,"claim":"Demonstrating that KIF11 physically binds ZBP1 and transports β-actin mRNA to cell protrusions established a non-mitotic, mRNA-transport function for this motor, linking it to cell polarity and migration.","evidence":"Reciprocal co-IP, RNA immunoprecipitation, smFISH colocalization, shRNA knockdown with rescue, migration assays","pmids":["25588836"],"confidence":"High","gaps":["Whether other mRNAs are KIF11 cargo","Structural basis for KIF11–ZBP1 interaction"]},{"year":2015,"claim":"Single-molecule reconstitution showed TPX2 directly inhibits Eg5 velocity through both motor interaction and microtubule binding, resolving the mechanism by which TPX2 dampens Eg5-dependent forces in the spindle.","evidence":"TIRF single-molecule motility and microtubule gliding assays with full-length and truncated TPX2","pmids":["26018074"],"confidence":"High","gaps":["Quantitative contribution of TPX2 inhibition to spindle force balance in vivo","Which TPX2 residues mediate the interaction"]},{"year":2016,"claim":"Identification of RNF20/40-mediated monoubiquitination as a stabilizing modification for Eg5 during mitosis, and Aurora A–Kif15 as a parallel spindle pathway, expanded the regulatory landscape beyond phosphorylation and defined genetic determinants of Eg5-independent spindle assembly.","evidence":"In vitro ubiquitination assays, co-IP, and genome-wide siRNA screen in Eg5-independent cells with Aurora A kinase validation","pmids":["27557628","27354041"],"confidence":"High","gaps":["Ubiquitination site(s) on Eg5 not mapped","How cells switch between Eg5-dependent and Kif15-dependent modes"]},{"year":2017,"claim":"Discovery that HDAC1 deacetylates Eg5 to regulate its ATPase activity, and that PP2A/B55α dephosphorylates Thr926 to enable mitotic exit, defined the enzymatic erasers that reset Eg5 post-translational modifications after mitotic entry.","evidence":"HDAC1 substrate trapping, ATPase assays, phosphatase assays, phosphomimetic mutant live imaging","pmids":["28392145","28487562"],"confidence":"High","gaps":["Specific acetylation site(s) on Eg5 not identified","Whether other phosphatases contribute at different mitotic stages"]},{"year":2018,"claim":"Multiple studies extended KIF11 function to postmitotic neurons: NEK7 phosphorylation promotes Eg5-dependent dendrite morphogenesis by restraining retrograde microtubule polymerization, and KIF11 knockdown increases excitatory synaptic transmission, revealing a synaptic inhibitory role.","evidence":"In vitro kinase assays, neuronal RNAi/KO in vivo, electrophysiology in hippocampal neurons","pmids":["29899413","30479371"],"confidence":"High","gaps":["Mechanism linking KIF11 to synaptic vesicle release or receptor density","Whether motor activity or microtubule-organizing function mediates the synaptic phenotype"]},{"year":2021,"claim":"Demonstration that Eg5 and PRC1-dependent KIF4A constitute two mechanistically distinct antiparallel microtubule sliding modules during anaphase B resolved how chromosome segregation proceeds through redundant force generators.","evidence":"Combined siRNA/pharmacological/CRISPR disruption with tubulin photoactivation and STED/expansion microscopy","pmids":["33910056"],"confidence":"High","gaps":["How the two modules are temporally coordinated","Whether additional sliding motors contribute"]},{"year":2023,"claim":"Finding that cytoplasmic NEAT1 lncRNA binds KIF11 and promotes its degradation to induce senescence via CDKN2A upregulation identified a non-coding RNA-based mechanism for KIF11 turnover outside of mitosis.","evidence":"RNA pulldown, mass spectrometry, co-IP, RNA FISH, ChIP, mouse xenograft model","pmids":["37752791"],"confidence":"High","gaps":["Degradation pathway (proteasomal vs lysosomal) not defined","Whether NEAT1-mediated KIF11 degradation occurs in non-tumor cell senescence"]},{"year":null,"claim":"Key unresolved questions include the structural basis for KIF11 tetramer-mediated antiparallel microtubule sliding at atomic resolution, the identity of specific acetylation sites modulated by HDAC1, the full cargo repertoire of KIF11 in mRNA transport, and how motor versus non-motor activities of KIF11 are partitioned in postmitotic neurons.","evidence":"","pmids":[],"confidence":"Low","gaps":["No high-resolution cryo-EM structure of full-length tetrameric KIF11 on microtubules","Acetylation site mapping incomplete","Complete mRNA cargo inventory lacking"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140657","term_label":"ATP-dependent activity","supporting_discovery_ids":[4]},{"term_id":"GO:0003774","term_label":"cytoskeletal motor activity","supporting_discovery_ids":[4,14]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[1,2]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[13]}],"localization":[{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[1,3,14]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,9,10,28]},{"term_id":"GO:0005829","term_label":"cytosol","supporting_discovery_ids":[13,27]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,3,9,10,15,22]},{"term_id":"R-HSA-112316","term_label":"Neuronal System","supporting_discovery_ids":[19,23]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[15,17]}],"complexes":["Eg5 homotetramer","TPX2–Eg5 inhibitory complex","ZBP1–β-actin mRNP"],"partners":["TPX2","KIF15","ZBP1","HMMR","RNF20","PTEN","HDAC1","NEK7"],"other_free_text":[]},"mechanistic_narrative":"KIF11 (Eg5) is a plus-end-directed kinesin-5 motor essential for centrosome separation, bipolar mitotic spindle assembly, and spindle elongation during cell division. Its spindle recruitment is controlled by CDK1 phosphorylation at Thr926/927 and Nek6/7 phosphorylation at Ser1033 downstream of a Plk1–Nek9 cascade, while PP2A/B55α dephosphorylates Thr926 to promote mitotic exit; additional post-translational regulation includes HDAC1-mediated deacetylation that modulates ATPase activity and RNF20/40-dependent monoubiquitination that stabilizes the protein [PMID:7753799, PMID:21642957, PMID:21522128, PMID:28487562, PMID:28392145, PMID:27557628]. KIF11 sliding forces are functionally balanced by the minus-end motor HSET, opposed by Tiam1–Rac1 signaling, dampened by HMMR-mediated TPX2 inhibitory complexes, and supplemented by a parallel Kif15–TPX2 sliding module and a PRC1–KIF4A module during anaphase [PMID:10525540, PMID:20346677, PMID:29386294, PMID:19818618, PMID:33910056]. Beyond mitosis, KIF11 transports β-actin mRNA in a ZBP1-dependent manner to regulate cell polarity and migration, promotes dendrite morphogenesis via NEK7 phosphorylation-dependent microtubule regulation, and acts as an inhibitory constraint on excitatory synaptic transmission in hippocampal neurons [PMID:25588836, PMID:29899413, PMID:30479371]."},"prefetch_data":{"uniprot":{"accession":"P52732","full_name":"Kinesin-like protein KIF11","aliases":["Kinesin-like protein 1","Kinesin-like spindle protein HKSP","Kinesin-related motor protein Eg5","Thyroid receptor-interacting protein 5","TR-interacting protein 5","TRIP-5"],"length_aa":1056,"mass_kda":119.2,"function":"Motor protein required for establishing a bipolar spindle and thus contributing to chromosome congression during mitosis (PubMed:19001501, PubMed:37728657). Required in non-mitotic cells for transport of secretory proteins from the Golgi complex to the cell surface (PubMed:23857769)","subcellular_location":"Cytoplasm; Cytoplasm, cytoskeleton, spindle pole","url":"https://www.uniprot.org/uniprotkb/P52732/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/KIF11","classification":"Common Essential","n_dependent_lines":1205,"n_total_lines":1208,"dependency_fraction":0.9975165562913907},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000138160","cell_line_id":"CID001413","localizations":[{"compartment":"cytoplasmic","grade":3},{"compartment":"cytoskeleton","grade":3}],"interactors":[{"gene":"PDZD8","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID001413","total_profiled":1310},"omim":[{"mim_id":"617569","title":"KINESIN FAMILY, MEMBER 15; KIF15","url":"https://www.omim.org/entry/617569"},{"mim_id":"614642","title":"START 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Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/33995648","citation_count":17,"is_preprint":false},{"pmid":"39806429","id":"PMC_39806429","title":"Identification and validation of the important role of KIF11 in the development and progression of endometrial cancer.","date":"2025","source":"Journal of translational medicine","url":"https://pubmed.ncbi.nlm.nih.gov/39806429","citation_count":16,"is_preprint":false},{"pmid":"33554761","id":"PMC_33554761","title":"High KIF11 expression is associated with poor outcome of NSCLC.","date":"2021","source":"Tumori","url":"https://pubmed.ncbi.nlm.nih.gov/33554761","citation_count":16,"is_preprint":false},{"pmid":"36513626","id":"PMC_36513626","title":"Kinesin-5 Eg5 is essential for spindle assembly, chromosome stability and organogenesis in development.","date":"2022","source":"Cell death discovery","url":"https://pubmed.ncbi.nlm.nih.gov/36513626","citation_count":16,"is_preprint":false},{"pmid":"28487562","id":"PMC_28487562","title":"Protein Phosphatase 2A (PP2A) Regulates EG5 to Control Mitotic Progression.","date":"2017","source":"Scientific reports","url":"https://pubmed.ncbi.nlm.nih.gov/28487562","citation_count":16,"is_preprint":false},{"pmid":"36051882","id":"PMC_36051882","title":"Using a comprehensive approach to investigate the interaction between Kinesin-5/Eg5 and the microtubule.","date":"2022","source":"Computational and structural biotechnology journal","url":"https://pubmed.ncbi.nlm.nih.gov/36051882","citation_count":16,"is_preprint":false},{"pmid":"34504599","id":"PMC_34504599","title":"ASPM combined with KIF11 promotes the malignant progression of hepatocellular carcinoma via the Wnt/β-catenin signaling pathway.","date":"2021","source":"Experimental and therapeutic medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34504599","citation_count":16,"is_preprint":false},{"pmid":"35140744","id":"PMC_35140744","title":"LncRNA VPS9D1-AS1 Promotes Malignant Progression of Lung Adenocarcinoma by Targeting miRNA-30a-5p/KIF11 Axis.","date":"2022","source":"Frontiers in genetics","url":"https://pubmed.ncbi.nlm.nih.gov/35140744","citation_count":16,"is_preprint":false},{"pmid":"18591782","id":"PMC_18591782","title":"Cloning, enzyme characterization of recombinant human Eg5 and the development of a new inhibitor.","date":"2008","source":"Biological & pharmaceutical bulletin","url":"https://pubmed.ncbi.nlm.nih.gov/18591782","citation_count":16,"is_preprint":false},{"pmid":"27146033","id":"PMC_27146033","title":"In Vitro Maturation of Mouse Oocytes Increases the Level of Kif11/Eg5 on Meiosis II Spindles.","date":"2016","source":"Biology of reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/27146033","citation_count":16,"is_preprint":false},{"pmid":"28965307","id":"PMC_28965307","title":"The kinesin Eg5 inhibitor K858 induces apoptosis and reverses the malignant invasive phenotype in human glioblastoma cells.","date":"2017","source":"Investigational new 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Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/28625631","citation_count":14,"is_preprint":false},{"pmid":"31428438","id":"PMC_31428438","title":"KIF11 microdeletion is associated with microcephaly, chorioretinopathy and intellectual disability.","date":"2018","source":"Human genome variation","url":"https://pubmed.ncbi.nlm.nih.gov/31428438","citation_count":14,"is_preprint":false},{"pmid":"32165913","id":"PMC_32165913","title":"Kinesin-5 Eg5 is essential for spindle assembly and chromosome alignment of mouse spermatocytes.","date":"2020","source":"Cell division","url":"https://pubmed.ncbi.nlm.nih.gov/32165913","citation_count":14,"is_preprint":false},{"pmid":"36929571","id":"PMC_36929571","title":"Morphological Profiling Identifies the Motor Protein Eg5 as Cellular Target of Spirooxindoles.","date":"2023","source":"Angewandte Chemie (International ed. in English)","url":"https://pubmed.ncbi.nlm.nih.gov/36929571","citation_count":14,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":46337,"output_tokens":7062,"usd":0.12247},"stage2":{"model":"claude-opus-4-6","input_tokens":10841,"output_tokens":3713,"usd":0.220545},"total_usd":0.343015,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 1995,\n      \"finding\": \"Eg5 (KIF11) spindle localization is cell-cycle-regulated and requires phosphorylation at a conserved threonine (T937) in the C-terminal domain; mutation of T937 to non-phosphorylatable alanine abolishes spindle localization, while T937S preserves it, implicating p34cdc2 phosphorylation as the regulatory mechanism.\",\n      \"method\": \"Transient transfection of myc-tagged Eg5 deletion/point mutants in Xenopus A6 cells; immunofluorescence localization\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — site-directed mutagenesis of candidate phosphorylation site with clear localization phenotype, replicated with multiple mutant constructs\",\n      \"pmids\": [\"7753799\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"Eg5 protein associates with both spindle and interphase microtubules during Xenopus early development, concentrating toward spindle poles during meiosis and mitosis; copellets with microtubules from egg homogenates, demonstrating direct microtubule association.\",\n      \"method\": \"Immunofluorescence and microtubule co-pelleting from Xenopus egg extracts; Western blotting across developmental stages\",\n      \"journal\": \"Developmental biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — biochemical co-sedimentation plus immunofluorescence in multiple developmental contexts\",\n      \"pmids\": [\"8026619\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1998,\n      \"finding\": \"Eg5 (KIF11) continues to be expressed in postmitotic rodent neurons, where it associates directly with the microtubule array and is enriched in distal regions of developing axons and dendrites, suggesting a role in regulating microtubule organization in neuronal processes.\",\n      \"method\": \"Immunofluorescence and subcellular fractionation in rodent neurons; expression profiling across developmental stages\",\n      \"journal\": \"The Journal of neuroscience\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct localization by immunofluorescence in postmitotic neurons with functional implication, single lab\",\n      \"pmids\": [\"9742151\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"HSET (minus-end motor) and Eg5 (KIF11, plus-end motor) act antagonistically in spindle assembly: simultaneous inhibition of both motors in cells restores centrosome separation and bipolar spindle formation that is blocked by Eg5 inhibition alone, establishing a balance-of-forces model for spindle bipolarity.\",\n      \"method\": \"Antibody microinjection in murine oocytes and cultured cells; immunoEM localization; in vitro aster assembly assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — reconstitution/in vitro assays combined with microinjection epistasis, replicated across multiple cell systems\",\n      \"pmids\": [\"10525540\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Monomeric human Eg5 is a slow plus-end-directed ATPase motor; kinetic analysis shows mantATP binds at 2–3 µM⁻¹s⁻¹, ATP hydrolysis occurs at 15 s⁻¹, and ATP-dependent microtubule dissociation is rate-limiting at ~8 s⁻¹, with mantADP release at 40 s⁻¹, distinguishing Eg5 mechanism from minus-end motors where ADP release is rate-limiting.\",\n      \"method\": \"In vitro ATPase kinetics (steady-state and transient), analytical ultracentrifugation, gel filtration, fluorescent nucleotide (mantATP/mantADP) binding assays with purified Eg5-367 and Eg5-437 constructs\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — rigorous in vitro reconstitution with multiple orthogonal kinetic methods on purified protein\",\n      \"pmids\": [\"15247293\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"TPX2 interacts with Eg5 via its C-terminal domain; this interaction is essential for Eg5-dependent spindle pole segregation, as TPX2-CT injection arrests spindle poles and Eg5 co-injection rescues cleavage, defining a discrete Eg5-interacting domain in TPX2.\",\n      \"method\": \"Blastomere microinjection in Xenopus embryos; cell transfection; co-immunoprecipitation; deletion analysis of TPX2\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — domain mapping combined with functional rescue experiments in two independent systems\",\n      \"pmids\": [\"18372177\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Parkin (E3 ubiquitin ligase) represses Eg5 transcription by causing multiple monoubiquitination of Hsp70, which inactivates JNK, reducing c-Jun phosphorylation and thereby blocking c-Jun binding to the AP-1 site in the Eg5 promoter; this is not via proteasomal Eg5 degradation.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assays, Eg5 promoter-reporter assay, ChIP, siRNA knockdown, Western blotting\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods (ChIP, reporter assay, ubiquitination assay) establishing transcriptional regulatory mechanism\",\n      \"pmids\": [\"18845538\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Kif15 (kinesin-12) cooperates with Eg5 to promote bipolar spindle assembly; Kif15 can fully replace Eg5 for centrosome separation when overexpressed, and this activity requires both its motor domain and interaction with the microtubule-associated protein TPX2, suggesting a Kif15–TPX2 complex slides antiparallel microtubules apart.\",\n      \"method\": \"siRNA depletion, ectopic overexpression, live-cell imaging, co-immunoprecipitation in human cell lines\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — epistasis plus interaction mapping, multiple orthogonal methods, replicated across conditions\",\n      \"pmids\": [\"19818618\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Tiam1–Rac1 signaling at centrosomes during prophase functions as the first identified antagonist of Eg5-driven centrosome separation; Tiam1/Rac1 depletion allows cells to escape Eg5-inhibitor-induced mitotic arrest, and re-inhibition of Eg5 in Tiam1-depleted cells rescues chromosome congression errors, placing Tiam1–Rac upstream of Eg5 forces in bipolar spindle assembly.\",\n      \"method\": \"siRNA depletion in culture cells, live-cell imaging, Rac1-deficient epithelial cells in vivo, epistasis double-inhibition experiments\",\n      \"journal\": \"Current biology : CB\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis in vitro and in vivo with multiple orthogonal approaches\",\n      \"pmids\": [\"20346677\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"CDK1 phosphorylates Eg5 at Thr926 (T927), and Plk1 controls centrosome separation additionally through activation of the Nek9–Nek6/7 kinase cascade, which phosphorylates Eg5 at Ser1033; both CDK1 (Thr926) and Nek6/7 (Ser1033) phosphorylation sites contribute to Eg5 accumulation at centrosomes and are necessary for centrosome separation.\",\n      \"method\": \"In vitro kinase assays, phospho-specific antibodies, siRNA/inhibitor epistasis, immunofluorescence in human cell lines\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro kinase assays plus cellular epistasis, two independent sites characterized\",\n      \"pmids\": [\"21642957\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Plk1 controls Eg5 localization to the centrosome in G2 phase and triggers centrosome separation independently of CDK1; CDK1 promotes fast centrosome separation while Plk1-driven separation is slow and opposed by actin-dependent forces; Cdk2 can also phosphorylate Eg5 at Thr927.\",\n      \"method\": \"Plk1/CDK1/CDK2 inhibitors, RNAi, phospho-specific antibodies, live-cell imaging, pharmacological manipulation of microtubule and actin dynamics\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple kinase inhibitors/knockdowns with live imaging and phospho-antibody validation\",\n      \"pmids\": [\"21522128\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Dynein does not directly antagonize Eg5 via a simple push-pull mechanism at the spindle equator; instead, live-cell imaging shows dynein and Eg5 activities cannot be titrated against each other, indicating indirect antagonism at different spindle locations.\",\n      \"method\": \"Live-cell imaging, endpoint quantification controls, mathematical modeling, siRNA depletion of dynein/Lis1\",\n      \"journal\": \"Cell reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — live imaging plus mathematical modeling, single study\",\n      \"pmids\": [\"22832270\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Kinetochore-microtubule stability governs the requirement for Eg5 in metaphase bipolar spindle maintenance; cells with more stable K-fibers can maintain bipolar spindles without Eg5, while artificial destabilization of K-MTs promotes spindle collapse without Eg5, and stabilization rescues bipolarity.\",\n      \"method\": \"Sibling cell line comparison, pharmacological K-MT stabilization/destabilization, Eg5 inhibitor treatment, immunofluorescence quantification\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — pharmacological epistasis with quantitative phenotypic readouts, single lab\",\n      \"pmids\": [\"24807901\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"KIF11 physically interacts with ZBP1 (IGF2BP1) and is a component of β-actin mRNP complexes; KIF11 colocalizes with β-actin mRNA and its ability to transport β-actin mRNA is dependent on ZBP1; disruption of the KIF11–ZBP1 interaction in vivo delocalizes β-actin mRNA and impairs cell migration.\",\n      \"method\": \"Co-immunoprecipitation, RNA immunoprecipitation, in vitro binding assay with domain mapping, smFISH colocalization, shRNA knockdown with rescue, migration assay\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP, domain mapping, functional rescue, multiple orthogonal methods\",\n      \"pmids\": [\"25588836\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"TPX2 inhibits Eg5 motor activity through two mechanisms: direct interaction with Eg5 (requiring dimerization or the neck region) and binding to microtubules; full-length TPX2 dramatically reduces Eg5 velocity at single-molecule level, while truncated TPX2 lacking the Eg5-interaction domain is less inhibitory.\",\n      \"method\": \"TIRF single-molecule motility assays with mammalian cell extracts expressing Eg5-EGFP; microtubule gliding assays; truncation analysis of TPX2\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — single-molecule reconstitution with domain mutants, two orthogonal in vitro assays\",\n      \"pmids\": [\"26018074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The RNF20/40 ubiquitin ligase complex interacts with Eg5 during mitosis and monoubiquitinates Eg5, stabilizing it; loss of RNF20/40 causes spindle assembly defects, cell cycle arrest and apoptosis, and depletes Eg5 protein levels.\",\n      \"method\": \"Co-immunoprecipitation, in vitro ubiquitination assay, siRNA knockdown, Western blotting, spindle phenotype analysis\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro ubiquitination reconstitution plus cellular co-IP and functional phenotype, multiple methods\",\n      \"pmids\": [\"27557628\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"PTEN associates and co-localizes with EG5 during mitosis; PTEN deficiency causes aberrant EG5 phosphorylation and abrogates EG5 recruitment to the mitotic spindle, leading to shorter spindles and chromosome misalignment.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence co-localization, PTEN siRNA knockdown, phospho-EG5 Western blotting\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP, functional KD phenotype, phospho-specific antibodies, multiple methods\",\n      \"pmids\": [\"27492783\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"HDAC1 deacetylates Eg5 (KIF11); HDAC1 co-localizes with Eg5 during mitosis and influences its ATPase activity; HDAC1/2-selective inhibition causes mitotic arrest and monopolar spindle formation consistent with Eg5 acetylation impairing function, identifying Eg5 as a non-histone HDAC1 substrate regulating mitotic progression.\",\n      \"method\": \"HDAC1 trapping mutant strategy to identify substrates, co-localization by immunofluorescence, ATPase activity assay, HDAC inhibitor treatment with spindle phenotype analysis\",\n      \"journal\": \"Cell chemical biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — substrate trapping, enzymatic assay, and functional cellular phenotype in one study\",\n      \"pmids\": [\"28392145\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"PP2A/B55α complex physically associates with the EG5 C-terminal tail domain and dephosphorylates EG5 at Thr926, enabling mitotic exit; PP2A knockdown leads to elevated phospho-EG5 in late metaphase and delayed mitotic exit, similar to EG5/T926D phosphomimetic mutant.\",\n      \"method\": \"Co-immunoprecipitation, in vitro phosphatase assay, phospho-specific antibodies, siRNA knockdown, EG5 phosphomimetic mutant transfection, live-cell imaging\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro phosphatase assay plus cellular genetics with phosphomimetic validation\",\n      \"pmids\": [\"28487562\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NEK7 phosphorylates Eg5/KIF11, promoting its accumulation on microtubules in distal dendrites; in this postmitotic context, Eg5 limits retrograde microtubule polymerization independently of its motor activity, thereby promoting dendrite growth and branching.\",\n      \"method\": \"In vitro kinase assay, RNAi knockdown in neurons (in vitro and in vivo), NEK7 kinase-dead mutant, pharmacological Eg5 inhibition, live imaging of microtubule dynamics\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro kinase assay plus neuronal KD/KO with functional phenotype, in vivo validation\",\n      \"pmids\": [\"29899413\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"HMMR dampens Eg5-mediated forces by localizing TPX2 and promoting formation of inhibitory TPX2–Eg5 complexes; HMMR silencing disturbs spindle microtubule organization and causes elevated aneuploidy, and Eg5 chemical inhibition rescues spindle defects in HMMR-depleted cells.\",\n      \"method\": \"siRNA silencing, co-immunoprecipitation, Eg5 inhibitor rescue, immunofluorescence, CRISPR deletion\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — co-IP combined with pharmacological rescue epistasis and CRISPR validation\",\n      \"pmids\": [\"29386294\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"HIV-1 Tat protein interacts with Eg5 and allosterically modulates its ATPase activity by affecting ADP release; lysine 85 in the Tat C-terminus is critical for this interaction and for Tat-induced mitotic arrest and apoptosis in CD4+ T lymphocytes.\",\n      \"method\": \"Co-immunoprecipitation, ATPase activity assay (ADP release kinetics), site-directed mutagenesis of Tat K85, siRNA, cell cycle/apoptosis analysis\",\n      \"journal\": \"The Journal of pathology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro ATPase assay with mutagenesis plus cellular functional phenotype\",\n      \"pmids\": [\"24488929\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"EG5/kinesin-5 together with PRC1-dependent KIF4A/kinesin-4 constitute two mechanistically distinct microtubule-sliding modules that drive spindle elongation in human cells; disruption of both EG5 and KIF4A blocks midzone microtubule sliding and causes total failure of chromosome segregation despite poleward chromosome motion.\",\n      \"method\": \"Combined siRNA depletion and pharmacological inactivation, CRISPR, tubulin photoactivation, STED microscopy, expansion microscopy\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple redundancy-testing approaches including CRISPR plus live imaging and super-resolution, rigorous controls\",\n      \"pmids\": [\"33910056\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"KIF11 knockdown in hippocampal neurons increases excitatory post-synaptic current frequency and amplitude, enhances dendritic arborization and synapse number, and requires pre-synaptic NMDAR activity for its effect; expression of Piccolo (active zone protein) constrains KIF11 function in synaptic transmission.\",\n      \"method\": \"RNAi-mediated knockdown, electrophysiological recordings (EPSCs) in hippocampal neurons, immunofluorescence morphometry, NMDAR pharmacological blockade\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — electrophysiology plus KD with pharmacological pathway dissection, single lab\",\n      \"pmids\": [\"30479371\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2007,\n      \"finding\": \"Homozygous knockout of Eg5 (Knsl1) causes early embryonic lethality prior to implantation in mice, with reduced cell numbers and failure of compaction; heterozygous mice are normal and fertile, establishing that Eg5 is essential for early mammalian cell division.\",\n      \"method\": \"Gene targeting (knockout mouse), embryo recovery at E2.5–3.5, morphological analysis\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean genetic knockout with defined developmental phenotype, confirmed by independent study (PMID 18474226)\",\n      \"pmids\": [\"17449012\", \"18474226\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Aurora A promotes bipolar spindle assembly in Eg5-independent cells by phosphorylating Kif15, enhancing Kif15 localization to the spindle; MCAK and Kif18b promote bipolar spindle assembly by destabilizing astral microtubules, placing Aurora A and Kif15 in a parallel pathway to Eg5 for centrosome separation.\",\n      \"method\": \"Genome-wide siRNA screen in Eg5-independent cells, Aurora A kinase assays, live-cell imaging, phospho-Kif15 antibodies\",\n      \"journal\": \"Chromosoma\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — genome-wide screen validated with kinase assay and live imaging, mechanistic pathway placement\",\n      \"pmids\": [\"27354041\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"Postnatal endothelial cell-specific conditional knockout of Kif11 in mice leads to severely stunted retinal vasculature growth and mildly stunted cerebellar vasculature, demonstrating that Kif11 function in mitotically active endothelial cells is most critical in rapidly growing CNS regions and that retinal hypovascularization can arise from impaired mitotic spindle function independently of Wnt/β-catenin signaling.\",\n      \"method\": \"Conditional knockout mouse (Cre-lox), retinal vascular whole-mount imaging, β-catenin signaling analysis\",\n      \"journal\": \"Human molecular genetics\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional KO in defined cell type with mechanistic pathway exclusion\",\n      \"pmids\": [\"31993640\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"NEAT1 lncRNA, when it translocates to the cytosol during senescence, interacts with KIF11 motor protein and promotes KIF11 protein degradation; KIF11 degradation leads to increased CDKN2A (p16/p14ARF) expression, inducing cellular senescence in hepatoma cells.\",\n      \"method\": \"RNA pulldown, mass spectrometry, co-immunoprecipitation, RNA FISH/immunofluorescence co-localization, ChIP, luciferase reporter assay, siRNA/shRNA knockdown, mouse xenograft model\",\n      \"journal\": \"Clinical and translational medicine\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — RNA pulldown + MS identification + co-IP validation + functional rescue with multiple orthogonal methods\",\n      \"pmids\": [\"37752791\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"PAK6 specifically co-localizes with Eg5 at the centrosome; PAK6 depletion upregulates Eg5 and causes multipolar spindles, while simultaneous Eg5 knockdown rescues this phenotype, establishing that PAK6 and Eg5 negatively inter-regulate each other to control spindle formation.\",\n      \"method\": \"Co-localization by immunofluorescence, siRNA knockdown, genetic epistasis (double knockdown rescue), Western blotting\",\n      \"journal\": \"Biochimica et biophysica acta. Molecular cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-localization and epistasis, single lab\",\n      \"pmids\": [\"33098954\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Death receptor 6 (DR6) physically interacts with KIF11, as confirmed by mass spectrometry co-purification, co-immunoprecipitation, and GST pull-down; overexpression of KIF11 eliminates suppression of ovarian cancer cell migration by DR6 knockdown, placing KIF11 downstream of DR6 in a pro-migratory signaling complex.\",\n      \"method\": \"Mass spectrometry, co-immunoprecipitation, GST pull-down, overexpression rescue assay, migration assay\",\n      \"journal\": \"FEBS open bio\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2–3 — MS identification confirmed by co-IP and pulldown plus functional rescue, single lab\",\n      \"pmids\": [\"30186750\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"KIF11/Eg5 is a homotetrameric, plus-end-directed kinesin-5 motor that crosslinks and slides antiparallel microtubules to drive centrosome separation and bipolar mitotic spindle assembly; its activity is tightly regulated by CDK1-mediated phosphorylation at Thr926/927 (which promotes spindle recruitment), Plk1/Nek9/Nek6-7 phosphorylation at Ser1033 (centrosome targeting), PP2A/B55α dephosphorylation (mitotic exit), HDAC1 deacetylation (ATPase regulation), and RNF20/40-dependent monoubiquitination (protein stabilization); it is opposed by HSET/minus-end motors, inhibited by TPX2 binding, and balanced by Kif15–TPX2 and other redundant sliding modules during spindle elongation; beyond mitosis, KIF11 mediates ZBP1-dependent β-actin mRNA transport to regulate cell polarity and migration, promotes dendrite morphogenesis via NEK7 phosphorylation, and functions as an inhibitory constraint on synaptic transmission in neurons.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"KIF11 (Eg5) is a plus-end-directed kinesin-5 motor essential for centrosome separation, bipolar mitotic spindle assembly, and spindle elongation during cell division. Its spindle recruitment is controlled by CDK1 phosphorylation at Thr926/927 and Nek6/7 phosphorylation at Ser1033 downstream of a Plk1–Nek9 cascade, while PP2A/B55α dephosphorylates Thr926 to promote mitotic exit; additional post-translational regulation includes HDAC1-mediated deacetylation that modulates ATPase activity and RNF20/40-dependent monoubiquitination that stabilizes the protein [PMID:7753799, PMID:21642957, PMID:21522128, PMID:28487562, PMID:28392145, PMID:27557628]. KIF11 sliding forces are functionally balanced by the minus-end motor HSET, opposed by Tiam1–Rac1 signaling, dampened by HMMR-mediated TPX2 inhibitory complexes, and supplemented by a parallel Kif15–TPX2 sliding module and a PRC1–KIF4A module during anaphase [PMID:10525540, PMID:20346677, PMID:29386294, PMID:19818618, PMID:33910056]. Beyond mitosis, KIF11 transports β-actin mRNA in a ZBP1-dependent manner to regulate cell polarity and migration, promotes dendrite morphogenesis via NEK7 phosphorylation-dependent microtubule regulation, and acts as an inhibitory constraint on excitatory synaptic transmission in hippocampal neurons [PMID:25588836, PMID:29899413, PMID:30479371].\",\n  \"teleology\": [\n    {\n      \"year\": 1994,\n      \"claim\": \"Establishing that Eg5 is a bona fide microtubule-associated motor present on spindles and interphase microtubules resolved the question of whether kinesin-5 family members physically engage the mitotic apparatus.\",\n      \"evidence\": \"Immunofluorescence and microtubule co-pelleting from Xenopus egg extracts across developmental stages\",\n      \"pmids\": [\"8026619\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Motor directionality and enzymatic mechanism not yet characterized\", \"No functional loss-of-function data\"]\n    },\n    {\n      \"year\": 1995,\n      \"claim\": \"Demonstrating that p34cdc2 (CDK1) phosphorylation of Thr937 is required for Eg5 spindle localization established the first regulatory switch controlling motor targeting to the mitotic apparatus.\",\n      \"evidence\": \"Site-directed mutagenesis of T937A/T937S in Xenopus A6 cells with immunofluorescence localization\",\n      \"pmids\": [\"7753799\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether additional kinases contribute to centrosome-specific targeting\", \"Phosphatase responsible for dephosphorylation unknown\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"Showing that simultaneous inhibition of HSET and Eg5 restores bipolarity established the balance-of-forces model, answering how opposing motors maintain spindle architecture.\",\n      \"evidence\": \"Antibody microinjection epistasis in murine oocytes and cultured cells combined with in vitro aster assembly\",\n      \"pmids\": [\"10525540\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether dynein contributes as a direct Eg5 antagonist\", \"Quantitative force contributions of each motor unknown\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Kinetic dissection of the Eg5 ATPase cycle revealed that ATP-dependent microtubule dissociation is rate-limiting, distinguishing Eg5 from conventional kinesin and explaining its slow processivity.\",\n      \"evidence\": \"Transient-state fluorescent nucleotide kinetics with purified monomeric Eg5 constructs\",\n      \"pmids\": [\"15247293\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How tetramerization alters the kinetic cycle\", \"No structural explanation for the rate-limiting step\"]\n    },\n    {\n      \"year\": 2007,\n      \"claim\": \"Homozygous Eg5 knockout causing pre-implantation lethality in mice definitively proved that Eg5 is essential for early mammalian cell division and cannot be compensated by other motors at this stage.\",\n      \"evidence\": \"Gene targeting (knockout mouse) with embryo recovery at E2.5–3.5\",\n      \"pmids\": [\"17449012\", \"18474226\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Cell-type-specific requirements in later development unexplored\", \"Whether Kif15 can partially rescue in specific lineages\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Identification of the TPX2 C-terminal domain as an Eg5-interacting element required for spindle pole separation, and of Parkin-mediated transcriptional repression of Eg5 via Hsp70 ubiquitination, defined two new layers of Eg5 regulation — one physical, one transcriptional.\",\n      \"evidence\": \"TPX2: co-IP and domain deletion with blastomere rescue in Xenopus; Parkin: ChIP, promoter-reporter, ubiquitination assays\",\n      \"pmids\": [\"18372177\", \"18845538\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether TPX2 inhibits Eg5 enzymatic activity directly (resolved later)\", \"Physiological contexts where Parkin regulation of Eg5 is relevant in vivo\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Discovery that Kif15 can fully substitute for Eg5 in centrosome separation when overexpressed revealed a redundant TPX2-dependent sliding pathway, explaining how some cancer cells escape Eg5 inhibitors.\",\n      \"evidence\": \"siRNA depletion and ectopic overexpression with live-cell imaging and co-IP in human cells\",\n      \"pmids\": [\"19818618\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Kif15 is regulated to become Eg5-independent\", \"Whether endogenous Kif15 levels ever suffice without overexpression\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Mapping dual phosphorylation inputs — CDK1 at Thr926 for spindle recruitment and Plk1→Nek9→Nek6/7 at Ser1033 for centrosome targeting — resolved how two independent kinase cascades converge to control Eg5 localization at distinct mitotic stages.\",\n      \"evidence\": \"In vitro kinase assays, phospho-specific antibodies, and siRNA/inhibitor epistasis in human cell lines\",\n      \"pmids\": [\"21642957\", \"21522128\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other phosphorylation sites contribute\", \"How actin-dependent forces oppose Plk1-driven slow separation\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Demonstrating that KIF11 physically binds ZBP1 and transports β-actin mRNA to cell protrusions established a non-mitotic, mRNA-transport function for this motor, linking it to cell polarity and migration.\",\n      \"evidence\": \"Reciprocal co-IP, RNA immunoprecipitation, smFISH colocalization, shRNA knockdown with rescue, migration assays\",\n      \"pmids\": [\"25588836\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether other mRNAs are KIF11 cargo\", \"Structural basis for KIF11–ZBP1 interaction\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Single-molecule reconstitution showed TPX2 directly inhibits Eg5 velocity through both motor interaction and microtubule binding, resolving the mechanism by which TPX2 dampens Eg5-dependent forces in the spindle.\",\n      \"evidence\": \"TIRF single-molecule motility and microtubule gliding assays with full-length and truncated TPX2\",\n      \"pmids\": [\"26018074\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quantitative contribution of TPX2 inhibition to spindle force balance in vivo\", \"Which TPX2 residues mediate the interaction\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identification of RNF20/40-mediated monoubiquitination as a stabilizing modification for Eg5 during mitosis, and Aurora A–Kif15 as a parallel spindle pathway, expanded the regulatory landscape beyond phosphorylation and defined genetic determinants of Eg5-independent spindle assembly.\",\n      \"evidence\": \"In vitro ubiquitination assays, co-IP, and genome-wide siRNA screen in Eg5-independent cells with Aurora A kinase validation\",\n      \"pmids\": [\"27557628\", \"27354041\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ubiquitination site(s) on Eg5 not mapped\", \"How cells switch between Eg5-dependent and Kif15-dependent modes\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Discovery that HDAC1 deacetylates Eg5 to regulate its ATPase activity, and that PP2A/B55α dephosphorylates Thr926 to enable mitotic exit, defined the enzymatic erasers that reset Eg5 post-translational modifications after mitotic entry.\",\n      \"evidence\": \"HDAC1 substrate trapping, ATPase assays, phosphatase assays, phosphomimetic mutant live imaging\",\n      \"pmids\": [\"28392145\", \"28487562\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Specific acetylation site(s) on Eg5 not identified\", \"Whether other phosphatases contribute at different mitotic stages\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Multiple studies extended KIF11 function to postmitotic neurons: NEK7 phosphorylation promotes Eg5-dependent dendrite morphogenesis by restraining retrograde microtubule polymerization, and KIF11 knockdown increases excitatory synaptic transmission, revealing a synaptic inhibitory role.\",\n      \"evidence\": \"In vitro kinase assays, neuronal RNAi/KO in vivo, electrophysiology in hippocampal neurons\",\n      \"pmids\": [\"29899413\", \"30479371\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism linking KIF11 to synaptic vesicle release or receptor density\", \"Whether motor activity or microtubule-organizing function mediates the synaptic phenotype\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Demonstration that Eg5 and PRC1-dependent KIF4A constitute two mechanistically distinct antiparallel microtubule sliding modules during anaphase B resolved how chromosome segregation proceeds through redundant force generators.\",\n      \"evidence\": \"Combined siRNA/pharmacological/CRISPR disruption with tubulin photoactivation and STED/expansion microscopy\",\n      \"pmids\": [\"33910056\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How the two modules are temporally coordinated\", \"Whether additional sliding motors contribute\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Finding that cytoplasmic NEAT1 lncRNA binds KIF11 and promotes its degradation to induce senescence via CDKN2A upregulation identified a non-coding RNA-based mechanism for KIF11 turnover outside of mitosis.\",\n      \"evidence\": \"RNA pulldown, mass spectrometry, co-IP, RNA FISH, ChIP, mouse xenograft model\",\n      \"pmids\": [\"37752791\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Degradation pathway (proteasomal vs lysosomal) not defined\", \"Whether NEAT1-mediated KIF11 degradation occurs in non-tumor cell senescence\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include the structural basis for KIF11 tetramer-mediated antiparallel microtubule sliding at atomic resolution, the identity of specific acetylation sites modulated by HDAC1, the full cargo repertoire of KIF11 in mRNA transport, and how motor versus non-motor activities of KIF11 are partitioned in postmitotic neurons.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No high-resolution cryo-EM structure of full-length tetrameric KIF11 on microtubules\", \"Acetylation site mapping incomplete\", \"Complete mRNA cargo inventory lacking\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140657\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0003774\", \"supporting_discovery_ids\": [4, 14]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [1, 2]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [13]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [1, 3, 14]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 9, 10, 28]},\n      {\"term_id\": \"GO:0005829\", \"supporting_discovery_ids\": [13, 27]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 3, 9, 10, 15, 22]},\n      {\"term_id\": \"R-HSA-112316\", \"supporting_discovery_ids\": [19, 23]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [15, 17]}\n    ],\n    \"complexes\": [\n      \"Eg5 homotetramer\",\n      \"TPX2–Eg5 inhibitory complex\",\n      \"ZBP1–β-actin mRNP\"\n    ],\n    \"partners\": [\n      \"TPX2\",\n      \"KIF15\",\n      \"ZBP1\",\n      \"HMMR\",\n      \"RNF20\",\n      \"PTEN\",\n      \"HDAC1\",\n      \"NEK7\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}