{"gene":"NUMA1","run_date":"2026-04-29T11:37:57","timeline":{"discoveries":[{"year":1996,"finding":"NuMA forms a complex with cytoplasmic dynein and dynactin; immunodepletion of NuMA from frog egg extracts abolishes normal spindle pole assembly, producing chromatin-associated irregular microtubule arrays; a NuMA tail subdomain induces microtubule aster formation by mediating microtubule bundling.","method":"Immunodepletion from Xenopus egg extracts, Co-immunoprecipitation, in vitro aster assembly assay","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1 — reconstitution in egg extracts with immunodepletion and rescue, replicated by multiple subsequent studies","pmids":["8898198"],"is_preprint":false},{"year":2000,"finding":"NuMA transport to spindle poles upon nuclear envelope breakdown is powered by cytoplasmic dynein and dynactin along microtubule minus ends; large cytoplasmic NuMA aggregates stream poleward in association with dynactin (Arp1) and dynein; immunoprecipitation and gel filtration demonstrate a reversible, mitosis-specific NuMA–dynein–dynactin complex; disruption of dynactin or dynein blocks NuMA translocation and spindle pole assembly.","method":"Live GFP imaging, immunoprecipitation, gel filtration, dynamitin overexpression, dynein antibody inhibition","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1–2 — multiple orthogonal methods in one study, consistent with prior immunodepletion data","pmids":["10811826"],"is_preprint":false},{"year":1992,"finding":"NuMA is a 236–238 kDa nuclear protein with a long central coiled-coil domain (~1,485 aa) flanked by globular N- and C-terminal domains; it dissociates from condensing chromosomes early in prophase before lamina disintegration and reassociates with telophase chromosomes before lamin accumulation, indicating a role in nuclear reformation.","method":"cDNA cloning, sequence analysis, double immunofluorescence with anti-NuMA and anti-lamin antibodies","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1 — primary structure determination with domain mapping, independently confirmed by Compton et al. 1992","pmids":["1541630","1541636"],"is_preprint":false},{"year":1995,"finding":"NuMA is required for organizing microtubules into aster-like mitotic arrays; immunodepletion of NuMA from mitotic cell extracts prevents aster formation, and addition of purified recombinant NuMA fully rescues aster assembly; NuMA is phosphorylated upon aster assembly and is only required in late stages of aster formation.","method":"Cell-free mitotic aster assembly assay, immunodepletion, recombinant protein rescue","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1 — reconstitution with immunodepletion and rescue by purified protein","pmids":["7593190"],"is_preprint":false},{"year":1995,"finding":"Mutation of the predicted CDK1 (p34cdc2) phosphorylation site at Thr2040 of NuMA (alone or combined with other site mutations) abolishes NuMA's ability to associate with spindle microtubules; instead the mutant concentrates at the plasma membrane, causing disorganized spindles, cytokinesis failure, and micronucleation. This demonstrates that mitosis-specific phosphorylation controls NuMA's spindle interaction.","method":"Site-directed mutagenesis, transient expression, immunofluorescence, flow cytometry","journal":"Journal of cell science","confidence":"High","confidence_rationale":"Tier 1 — mutagenesis with defined phosphorylation site linked to specific phenotype","pmids":["7769006"],"is_preprint":false},{"year":1993,"finding":"Expression of NuMA lacking its globular head domain results in cytokinesis failure and micronuclei formation; expression of NuMA lacking the globular tail domain prevents nuclear targeting and spindle binding, causing micronucleation in daughter cells. Wild-type NuMA overexpression rescues nuclear assembly defects in RCC1 temperature-sensitive cells. NuMA function is required for terminal phases of chromosome separation and/or nuclear reassembly.","method":"Dominant-negative expression, temperature-sensitive mutant cell rescue, immunofluorescence","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 — domain deletion mutants with well-defined phenotypes, rescue experiment","pmids":["8432734"],"is_preprint":false},{"year":2004,"finding":"LGN (mammalian Pins homolog) directly binds NuMA through its N-terminal domain and recruits NuMA to the cell cortex during mitosis; LGN behaves as a conformational switch—its closed state has N- and C-termini interacting, but NuMA or Gαi can open the switch, allowing simultaneous binding to both, resulting in their cortical localization. LGN–NuMA interaction is required for spindle oscillation.","method":"FRET biosensor, yeast two-hybrid, overexpression, RNAi, live imaging","journal":"Cell","confidence":"High","confidence_rationale":"Tier 1–2 — FRET structural insight plus functional rescue, multiple orthogonal approaches","pmids":["15537540"],"is_preprint":false},{"year":2001,"finding":"LGN (human Pins-related protein) binds the C-terminal tail of NuMA; LGN blocks NuMA-dependent stabilization and bundling of microtubules. In vitro assays show NuMA binds MTs directly, and the MT-binding domain overlaps by 10 aa with the LGN-binding domain, indicating steric exclusion explains LGN's inhibitory effect on NuMA spindle function.","method":"In vitro MT binding/stabilization assays, Xenopus egg extract aster assay, domain mapping","journal":"Nature cell biology / Current biology","confidence":"High","confidence_rationale":"Tier 1 — in vitro binding and MT stabilization assays with domain mutagenesis, two papers","pmids":["11781568","12445386"],"is_preprint":false},{"year":1999,"finding":"A 135 kDa nonerythroid isoform of protein 4.1R directly interacts with NuMA; minimal binding involves 4.1R exons 20–21 and NuMA residues 1788–1810; 4.1R and NuMA co-localize in interphase nuclei and redistribute to spindle poles, where 4.1R is part of a NuMA–dynein–dynactin complex during mitosis.","method":"Yeast two-hybrid, in vitro binding assay, co-immunoprecipitation, co-immunolocalization","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal binding methods with domain mapping, single lab","pmids":["10189366"],"is_preprint":false},{"year":2002,"finding":"NuMA contains an RXXPDG motif that mediates direct binding to the ankyrin-repeat domain of tankyrase-1 and tankyrase-2 (poly-ADP-ribose polymerases); this interaction contributes to the known co-localization of tankyrase and NuMA at mitotic spindle poles.","method":"Yeast two-hybrid, in vitro binding, co-localization","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 3 — yeast two-hybrid plus in vitro binding, single lab","pmids":["12080061"],"is_preprint":false},{"year":2005,"finding":"NuMA is a major acceptor of poly(ADP-ribosyl)ation (PARsylation) by tankyrase 1 in mitosis; tankyrase 1 and NuMA association increases at mitotic onset concomitant with NuMA PARsylation; siRNA knockdown of tankyrase 1 eliminates NuMA PARsylation; conversely, NuMA knockdown causes complete loss of tankyrase 1 from spindle poles.","method":"Co-immunoprecipitation, immunofluorescence, siRNA knockdown","journal":"The Biochemical journal","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP with siRNA rescue, single lab, single paper","pmids":["16076287"],"is_preprint":false},{"year":2009,"finding":"Poly(ADP-ribose) (pADPr) made by PARP-5a/tankyrase-1 localizes to spindle poles; pADPr-coated beads trigger microtubule aster assembly in mitotic HeLa lysate; the rod domain of NuMA binds directly to pADPr chains, suggesting pADPr provides dynamic cross-linking at spindle poles by binding NuMA.","method":"In vitro pADPr bead aster assay, recombinant domain binding, immuno-EM","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 1–2 — in vitro aster reconstitution plus direct binding, single lab","pmids":["19759176"],"is_preprint":false},{"year":1997,"finding":"NuMA is phosphorylated by CDK1 (Cdc2 kinase) in a mitosis-specific manner as shown in vivo with 32P-labeling; phosphorylation shifts NuMA to a higher apparent molecular weight form; this modification coincides with NuMA's release from the nucleus and redistribution to the spindle.","method":"32P metabolic labeling, phosphatase treatment, immunoblot in synchronized cells","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — biochemical characterization in synchronized cells with phosphatase controls, single lab","pmids":["9202389"],"is_preprint":false},{"year":2004,"finding":"Cyclin B degradation at anaphase onset leads to NuMA dephosphorylation and its release from dynein, dynactin, and spindle poles; a non-degradable cyclin B (Δ90) keeps NuMA phosphorylated and stably associated with dynein/dynactin at persistent spindle poles that fail to disassemble.","method":"Xenopus egg extract, non-degradable cyclin B expression, co-immunoprecipitation, immunofluorescence","journal":"EMBO reports","confidence":"Medium","confidence_rationale":"Tier 2 — biochemical rescue experiment with defined cyclin B mutant, single lab","pmids":["14710193"],"is_preprint":false},{"year":2013,"finding":"CDK1 phosphorylates NuMA at T2055 to negatively regulate its cortical localization during metaphase; PPP2CA (PP2A) phosphatase counteracts this; CDK1 inactivation in anaphase increases dephosphorylated cortical NuMA, enriching cortical dynein and driving spindle elongation. Thus, NuMA phosphorylation status by CDK1 couples mitotic progression with spindle behavior.","method":"In vitro kinase assay, phosphorylation-site mutagenesis, immunofluorescence, siRNA knockdown in human cells","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 — direct kinase assay with mutagenesis, functional rescue, single lab with multiple orthogonal methods","pmids":["23921553"],"is_preprint":false},{"year":2013,"finding":"A lipid/membrane-binding domain at the C-terminus of NuMA mediates LGN/Gαi-independent cortical association during anaphase; this binding is inhibited by CDK1 phosphorylation during prophase and metaphase and activated upon CDK1 inactivation at anaphase; replacing endogenous NuMA with membrane-binding-deficient NuMA reduces anaphase cortical dynein and impairs chromosome separation.","method":"Domain mapping, phosphorylation-site mutagenesis, live cell imaging, siRNA rescue experiments, lipid-binding assays","journal":"Molecular biology of the cell","confidence":"High","confidence_rationale":"Tier 1–2 — domain-function dissection with phosphomutants and rescue, multiple methods","pmids":["24371089"],"is_preprint":false},{"year":2014,"finding":"NuMA directly associates with phosphoinositides PtdInsP (PIP) and PtdInsP2 (PIP2) in vitro; depletion of PIP/PIP2 by chemical or enzymatic means prevents NuMA cortical localization during mitosis; increasing PIP2 augments cortical NuMA. During anaphase, LGN/Gαi are dispensable for NuMA cortical enrichment but PIP/PIP2 are required.","method":"In vitro lipid-binding assay, chemical/enzymatic PIP depletion, immunofluorescence, siRNA knockdown","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 — direct biochemical binding plus functional depletion with multiple rescue strategies","pmids":["24996901"],"is_preprint":false},{"year":2016,"finding":"Aurora-A directly phosphorylates the C-terminus of NuMA on three serine residues, including Ser1969; this phosphorylation governs the dynamic exchange of NuMA between cytoplasm/spindle poles and cortex; partial Aurora-A inhibition traps NuMA at spindle poles and prevents cortical enrichment. Aurora-A phosphorylation of NuMA does not affect LGN or MT binding affinity but controls NuMA mobility at poles.","method":"In vitro kinase assay, phospho-site mutagenesis, FRAP, immunofluorescence, kinase inhibition in human cells","journal":"Current biology","confidence":"High","confidence_rationale":"Tier 1 — in vitro kinase assay with mutagenesis, FRAP, and functional phenotype rescue","pmids":["26832443"],"is_preprint":false},{"year":2018,"finding":"Plk1 directly interacts with and phosphorylates NuMA; acute Plk1 inactivation enriches cortical NuMA/LGN/dynein during metaphase; Plk1-mediated NuMA phosphorylation controls NuMA cortical localization dynamics and spindle orientation.","method":"In vitro kinase assay, co-immunoprecipitation, pharmacological Plk1 inhibition, FRAP, immunofluorescence","journal":"Life science alliance","confidence":"Medium","confidence_rationale":"Tier 1–2 — in vitro kinase assay plus functional readout, single lab","pmids":["30456393"],"is_preprint":false},{"year":2009,"finding":"NuMA is an essential tether linking bulk spindle microtubules to centrosomes; conditional loss of NuMA function in mice shows that centrosomes provide initial spindle focusing but centrosome-spindle attachment under tension fails without NuMA, and kinetochore fiber maintenance at poles is lost; without both centrosomes and NuMA, all spindle focusing fails.","method":"Conditional loss-of-function allele (mouse genetics), primary cell culture, immunofluorescence, live imaging","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 — clean conditional KO with defined cellular and structural phenotypes, in vivo","pmids":["19255246"],"is_preprint":false},{"year":1999,"finding":"NuMA self-assembles in vitro into multiarm oligomers (up to 10–12 arms) through C-terminal globular domain interactions; each arm corresponds to a NuMA dimer; electron microscopy and computer modeling show these 12-arm oligomers are the structural units of a quasi-hexagonal nuclear scaffold observed upon NuMA overexpression in HeLa cells.","method":"In vitro assembly, electron microscopy, computer modeling, overexpression in HeLa cells","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution of NuMA oligomers with EM structural validation","pmids":["10075938"],"is_preprint":false},{"year":1995,"finding":"The NuMA rod domain forms a 207 nm long parallel, in-register double-stranded coiled-coil (the longest known); electron microscopy of purified recombinant fragments and full-length protein confirms tripartite structure with flanking globular domains; the SPN-3 antibody epitope maps to residues 255–267 in the first helical subdomain.","method":"Electron microscopy of recombinant protein, chemical cross-linking, circular dichroism, epitope mapping","journal":"The EMBO journal","confidence":"High","confidence_rationale":"Tier 1 — direct structural characterization of purified protein by EM and biophysical methods","pmids":["7781599"],"is_preprint":false},{"year":2017,"finding":"NuMA recruits dynactin to microtubule minus-ends; NuMA localizes to new minus-ends independently of dynein and faster than dynactin; a C-terminal region outside NuMA's canonical MT-binding domain mediates minus-end localization independently of γ-TuRC, CAMSAP1, or KANSL1/3; both NuMA's minus-end-binding and dynein-dynactin-binding modules are required to rescue bipolar spindle organization.","method":"Quantitative live imaging, laser ablation, domain deletion mutants, siRNA knockdown with rescue","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — quantitative imaging with laser ablation and domain-specific rescue, multiple orthogonal approaches","pmids":["29185983"],"is_preprint":false},{"year":2018,"finding":"Cortical targeting of NuMA (but not dynein alone) is sufficient to generate spindle-pulling forces; NuMA assembles specialized focal structures at the cortex that cluster multiple dynein-dynactin force-generating modules; NuMA's N-terminal long arm (dynein-dynactin recruitment), dynein-based MT gliding, and direct MT-binding activity are all required; NuMA clustering is essential for spindle positioning but not spindle-pole focusing.","method":"Light-induced cortical targeting (optogenetics/reconstitution), AID-based depletion, live imaging, domain dissection","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1–2 — light-induced reconstitution system with domain dissection, multiple orthogonal methods","pmids":["29848445"],"is_preprint":false},{"year":2011,"finding":"Crystal structures of LGN/NuMA and LGN/mInsc complexes reveal that NuMA and Inscuteable (mInsc) interact with the same LGN TPR domain in a mutually exclusive manner; mInsc binds with higher affinity; biochemical competition assays confirm this exclusivity, suggesting Par3/mInsc/LGN and NuMA/LGN/Gαi complexes play sequential rather than simultaneous roles.","method":"X-ray crystallography, in vitro competition binding assays, cell biology studies","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1 — crystal structures of both complexes with orthogonal biochemical validation","pmids":["21816348"],"is_preprint":false},{"year":2005,"finding":"Ric-8A, a Gαi guanine nucleotide exchange factor, catalytically dissociates Gαi-GDP/LGN/NuMA complexes in vitro by releasing activated Gαi-GTP, which concomitantly liberates NuMA from LGN; this mechanism may regulate microtubule pulling forces on centrosomes during cell division.","method":"In vitro GEF assay with purified components, biochemical complex dissociation assay","journal":"PNAS","confidence":"Medium","confidence_rationale":"Tier 1 — in vitro reconstitution assay with purified proteins, single lab","pmids":["16275912"],"is_preprint":false},{"year":2010,"finding":"Ric-8A and Gαi recruit LGN, NuMA, and dynein to the cell cortex during mitosis to orient the mitotic spindle; pertussis toxin (blocking Ric-8A GEF activity for Gαi), Ric-8A siRNA, or Gαi siRNA each impair cortical LGN, NuMA, and dynein localization and disturb integrin-dependent spindle orientation.","method":"siRNA knockdown, pertussis toxin treatment, live GFP-tubulin imaging, immunofluorescence","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 — multiple loss-of-function approaches with defined localization and orientation phenotype, single lab","pmids":["20479129"],"is_preprint":false},{"year":2014,"finding":"CYLD deubiquitinase deubiquitinates the cortical polarity protein dishevelled, enhancing dishevelled–NuMA interaction and promoting cortical localization of NuMA and the dynein/dynactin complex; CYLD also stabilizes astral microtubules to regulate spindle orientation.","method":"Co-immunoprecipitation, ubiquitination assay, siRNA knockdown, immunofluorescence","journal":"PNAS","confidence":"Medium","confidence_rationale":"Tier 2 — biochemical deubiquitination assay plus co-IP and loss-of-function, single lab","pmids":["24469800"],"is_preprint":false},{"year":2013,"finding":"NuMA's 4.1-binding domain is required for stabilizing NuMA at the cell cortex of keratinocytes (shown by FRAP); loss of 4.1/NuMA interaction causes spindle orientation defects; NuMA is required to recruit dynactin to the cortex; at anaphase, a CDK1-independent, LGN/4.1-independent mechanism also drives cortical NuMA accumulation.","method":"FRAP, domain mutant expression, siRNA knockdown, immunofluorescence in primary keratinocytes","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2 — FRAP with domain mutants and functional spindle orientation readout, single lab","pmids":["24109598"],"is_preprint":false},{"year":2016,"finding":"NuMA's direct MT-binding domain (targeting MT tips) is essential for spindle orientation establishment in keratinocytes; dynein/dynactin alone is insufficient; loss of NuMA–MT interactions in mouse skin causes spindle orientation defects, epidermal differentiation failure, and neonatal lethality; in adult mice, loss of NuMA-MT interactions disrupts hair follicle morphogenesis.","method":"MT-binding domain mutagenesis, conditional knockout in mouse skin, live imaging, immunofluorescence","journal":"eLife","confidence":"High","confidence_rationale":"Tier 2 — in vivo conditional KO with domain-specific mutagenesis, defined in vivo phenotype","pmids":["26765568"],"is_preprint":false},{"year":2017,"finding":"Crystal structure of Importin-α bound to the NuMA C-terminus reveals a novel NLS binding pattern enabling selective NLS recognition; Importin-β, in the presence of Importin-α, sterically masks a high-affinity MT-binding region of NuMA C-terminal to the NLS, thereby inhibiting NuMA's MT-bundling activity; RanGTP releases this inhibition.","method":"X-ray crystallography, in vitro MT-binding assay, Importin-β binding assay, mutagenesis","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1 — crystal structure with functional in vitro validation by MT-binding assay and mutagenesis","pmids":["28939615"],"is_preprint":false},{"year":2020,"finding":"NuMA contains two dynein LIC-binding sites: a hook domain contacting LIC1/LIC2 through a conserved hydrophobic patch (shared with Hook adaptors) and a CC1-box-like motif within its coiled-coil; both LIC-binding sites are essential for correct spindle positioning and cell division.","method":"X-ray crystallography of NuMA–LIC1/2 interfaces, mutagenesis, functional cell division assay","journal":"Structure","confidence":"High","confidence_rationale":"Tier 1 — crystal structure with mutagenesis and functional validation","pmids":["32413290"],"is_preprint":false},{"year":1999,"finding":"Once NuMA is incorporated into the insoluble spindle pole matrix in vivo or in vitro, its insolubility becomes independent of microtubules; immunogold EM shows NuMA at an electron-dense material distinct from pericentriolar material that appears to anchor microtubule ends; NuMA is essential for formation of this insoluble matrix.","method":"Immunogold electron microscopy, biochemical fractionation, immunodepletion of cell-free extracts","journal":"Cell motility and the cytoskeleton","confidence":"Medium","confidence_rationale":"Tier 2 — EM localization with biochemical fractionation and immunodepletion, single lab","pmids":["10098933"],"is_preprint":false},{"year":2006,"finding":"Rae1 (mRNA export factor) has a mitosis-specific interaction with NuMA; Rae1 binds a specific site on NuMA that would convert a NuMA dimer to a tetravalent MT crosslinker; reducing Rae1 or increasing NuMA disrupts spindle architecture; co-depletion or co-overexpression of Rae1 with NuMA prevents aberrant spindle formation.","method":"Co-immunoprecipitation, domain mapping, overexpression, siRNA knockdown in HeLa cells","journal":"PNAS","confidence":"Medium","confidence_rationale":"Tier 2 — reciprocal co-IP with domain mapping and epistasis by double perturbation, single lab","pmids":["17172455"],"is_preprint":false},{"year":2004,"finding":"NuMA dynamics at spindle poles involve continuous exchange between soluble and spindle-associated pools (t½ ~3 min by FRAP); this exchange requires cellular energy; LGN binding and protein kinase activity both regulate NuMA's dynamic exchange rate on spindle pole asters in cell-free extracts.","method":"FRAP in live cells, cell-free aster exchange assay, LGN addition, kinase inhibitors","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — FRAP plus cell-free assay with pharmacological and protein perturbations, single lab","pmids":["15561764"],"is_preprint":false},{"year":2021,"finding":"NuMA undergoes liquid-liquid phase separation (LLPS) at mitotic entry; this is regulated by Aurora-A phosphorylation; KifC1 facilitates NuMA condensate concentration at spindle poles; phase separation is mediated by NuMA's C-terminus and dynein-dynactin binding motif; phase-separated NuMA concentrates tubulins, binds MTs, and enriches Kif2A (a spindle MT depolymerizer) at poles to promote poleward MT flux.","method":"In vitro phase separation assay, live cell imaging of condensates, siRNA knockdown, phosphomimetic/phosphodead mutants","journal":"Nature communications","confidence":"Medium","confidence_rationale":"Tier 2 — in vitro LLPS plus live cell assays, single lab, novel mechanism","pmids":["34887424"],"is_preprint":false},{"year":2014,"finding":"NuMA accumulates at DNA damage sites in a poly-ADP-ribosylation (PARylation)-dependent manner; NuMA co-immunoprecipitates with the chromatin remodeler SNF2h/SMARCA5 and regulates SNF2h diffusion and its accumulation at DNA breaks; NuMA knockdown impairs chromatin decompaction after DNA cleavage, reduces homologous recombination repair factor recruitment, and impairs DSB repair in chromosomal (not episomal) contexts.","method":"Co-immunoprecipitation, FRAP, siRNA knockdown, laser microirradiation, DNA repair assays","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods establishing novel NuMA function in DNA repair, single lab","pmids":["24753406"],"is_preprint":false},{"year":2013,"finding":"NuMA binds p53 and is required for CDK8 (Mediator complex component) recruitment to p53 target gene promoters; acute NuMA knockdown selectively attenuates p21 induction after DNA damage (impairing cell cycle arrest) but does not affect PUMA induction, demonstrating NuMA confers selectivity to p53-mediated transcription.","method":"Co-immunoprecipitation, siRNA knockdown, chromatin immunoprecipitation, transcriptional reporter assays","journal":"Molecular and cellular biology","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP with ChIP and selective gene expression readout, single lab","pmids":["23589328"],"is_preprint":false},{"year":2019,"finding":"NuMA interacts with 53BP1 and controls 53BP1 diffusion throughout the nucleoplasm; this interaction is reduced after DNA damage; NuMA prevents 53BP1 accumulation at DNA breaks in the undamaged state; manipulating NuMA expression alters PARP inhibitor sensitivity in BRCA1-null cells and affects end-joining activity and immunoglobulin class switching.","method":"Co-immunoprecipitation, FRAP, siRNA knockdown, NHEJ assay, class-switch recombination assay","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal methods for NuMA–53BP1 interaction and functional readout, single lab","pmids":["30812030"],"is_preprint":false},{"year":2020,"finding":"In neurons, NuMA1 is transiently located at the axon initial segment (AIS) where it interacts with scaffolding protein 4.1B and the dynein regulator Lis1; NuMA1 inhibits endocytosis of AIS protein NF186 by impeding Lis1's interaction with doublecortin; silencing NuMA1 or 4.1B disrupts AIS assembly but not maintenance.","method":"Differential proteomics, co-immunoprecipitation, shRNA knockdown, endocytosis assay, immunofluorescence in mouse neurons","journal":"The Journal of cell biology","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP with mechanistic endocytosis assay and loss-of-function phenotype, single lab","pmids":["31727776"],"is_preprint":false},{"year":2021,"finding":"NUMA1 is downregulated in Huntington's disease neural progenitors by miR-124; suppression of NUMA1 in wild-type cells recapitulates HD microtubule bundling defects within axonal growth cones and impairs axon growth; restoring NUMA1 levels (via antagomiR-124) or stabilizing microtubules with epothilone B rescues microtubule organization and axonal growth.","method":"Proteomics of growth cones, miRNA manipulation, shRNA knockdown, rescue by antagomiR or drug, live imaging of growth cones","journal":"Neuron","confidence":"Medium","confidence_rationale":"Tier 2 — loss-of-function and rescue experiments with defined microtubule and axon growth phenotype, single lab","pmids":["34793694"],"is_preprint":false},{"year":2000,"finding":"GAS41 (a protein amplified in gliomas) directly binds the C-terminal rod region of NuMA (Kd ~2×10⁻⁷ M) as shown by dot overlay and surface plasmon resonance; GAS41 is nucleolar in interphase and becomes diffuse in mitosis.","method":"Yeast two-hybrid, dot overlay, surface plasmon resonance","journal":"The Journal of biological chemistry","confidence":"Low","confidence_rationale":"Tier 3 — in vitro binding without functional consequence established, single lab","pmids":["10913114"],"is_preprint":false},{"year":2017,"finding":"NuMA controls rDNA transcription and mediates nucleolar stress response in a p53-independent manner; NuMA co-immunoprecipitates with RNA Pol I, ribosomal proteins RPL26/RPL24, and B-WICH complex components; NuMA binds 18S and 28S rRNAs and localizes to rDNA promoter regions; NuMA knockdown decreases nascent pre-rRNA synthesis and triggers p27kip1 upregulation.","method":"Co-immunoprecipitation, RNA-binding assay, ChIP, siRNA knockdown, nascent RNA labeling","journal":"Nucleic acids research","confidence":"Medium","confidence_rationale":"Tier 2 — multiple binding and functional assays establishing novel nucleolar function, single lab","pmids":["28981686"],"is_preprint":false},{"year":2016,"finding":"NuMA directly binds Astrin (C-terminal tail of NuMA to C-terminus of Astrin); NuMA is required for Astrin recruitment to the mitotic spindle; LGN N-terminus (which blocks NuMA MT binding) also displaces Astrin from NuMA; dynein-mediated transport is required for spindle pole accumulation of Astrin; conversely, reduced Astrin impairs NuMA concentration at spindle poles.","method":"Co-immunoprecipitation, GST pulldown, siRNA knockdown, immunofluorescence","journal":"The Journal of biological chemistry","confidence":"Medium","confidence_rationale":"Tier 2 — direct binding assay plus reciprocal knockdown phenotypes, single lab","pmids":["27462074"],"is_preprint":false},{"year":1997,"finding":"NuMA is cleaved during apoptosis; the cleavage site lies between residues 1701 and 1725 generating a stable ~180–200 kDa fragment; cleavage is retarded by TPCK but not by ICE inhibitors or other protease inhibitors tested.","method":"Immunoblot, protease inhibitor treatment, apoptosis induction in cell lines","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 — cleavage site mapped with pharmacological dissection, single lab","pmids":["9184071"],"is_preprint":false},{"year":2003,"finding":"NuMA is preferentially cleaved by caspase-3 during Fas-mediated apoptosis; NuMA cleavage coincides with lamin B and PARP-1 cleavage; NuMA redistribution during apoptosis involves condensation, central nuclear concentration, and encirclement of apoptotic body fragments; NuMA was not cleaved in caspase-3-null MCF-7 cells treated with staurosporine.","method":"Immunofluorescence, immunoblot, caspase inhibitors, caspase-3-null cell line, Fas stimulation","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — genetic null cell line plus pharmacological inhibitors, single lab","pmids":["12508117"],"is_preprint":false},{"year":1994,"finding":"NuMA residues 1972–2007 constitute a nuclear localization signal (NLS); Lys1988 is essential for nuclear targeting; residues 1538–2115 are necessary and sufficient for spindle association; NuMA isoforms NuMA-m and NuMA-s (arising from alternative splicing) localize to centrosomes in interphase and spindle poles in mitosis rather than the nucleus.","method":"Linker scanning mutagenesis, chimeric protein expression, transfection in CHO cells, immunofluorescence","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — systematic domain mapping with point mutations and isoform analysis, single lab","pmids":["7962183"],"is_preprint":false},{"year":2017,"finding":"Galectin-3 associates with NuMA at spindle poles in an O-GlcNAcylation-dependent manner during metaphase; loss of Galectin-3 impairs spindle pole cohesion and stable NuMA localization at the spindle pole in epithelial cells.","method":"Co-immunoprecipitation, siRNA knockdown, immunofluorescence, experimental mouse model","journal":"Scientific reports","confidence":"Medium","confidence_rationale":"Tier 2 — co-IP with loss-of-function phenotype in both cell lines and mouse model, single lab","pmids":["28469279"],"is_preprint":false},{"year":2019,"finding":"In acentrosomal human cells, NuMA forms small microtubule asters at nuclear envelope breakdown that are assembled by dynein and NuMA's clustering activity; NuMA organizes radial MT arrays incorporating Eg5 to promote spindle bipolarization; in cells with centrosomes, NuMA also promotes the initial step of spindle bipolarization.","method":"siRNA depletion of centrosome components, live imaging, NuMA siRNA in acentrosomal cells, immunofluorescence","journal":"The EMBO journal","confidence":"Medium","confidence_rationale":"Tier 2 — defined genetic manipulation with live imaging in acentrosomal system, single lab","pmids":["31782546"],"is_preprint":false}],"current_model":"NuMA is a large coiled-coil nuclear protein that undergoes CDK1-mediated phosphorylation at mitotic entry to exit the nucleus and form a complex with cytoplasmic dynein and dynactin, which transports NuMA to spindle microtubule minus-ends where it acts as a minus-end cargo adaptor; at spindle poles NuMA self-assembles into an insoluble cross-linking matrix that focuses and anchors microtubule minus-ends, a process regulated by Aurora-A and Plk1 phosphorylation; at the cell cortex during metaphase, NuMA is recruited by the LGN/Gαi complex (itself activated by Ric-8A GEF activity) and directly by phosphoinositides, assembling dynein-dynactin clusters that generate pulling forces on astral microtubules to orient the spindle, with cortical NuMA levels precisely tuned by CDK1 (metaphase suppression), Aurora-A, Plk1, and PP2A activity; in anaphase, CDK1 inactivation allows LGN-independent, phosphoinositide-dependent cortical NuMA accumulation that drives spindle elongation; in interphase, NuMA resides in the nuclear matrix where it interacts with protein 4.1, p53, RNA Pol I, SNF2h, and 53BP1 to contribute to nuclear architecture, p53 target gene selectivity, rDNA transcription, chromatin remodeling at DNA breaks, and regulation of DNA repair pathway choice."},"narrative":{"teleology":[{"year":1992,"claim":"Cloning NuMA established it as a large nuclear protein with a tripartite domain architecture—globular heads flanking a long coiled-coil rod—that dynamically redistributes between the nucleus and spindle poles during mitosis, framing the central question of how a nuclear protein reorganizes into a mitotic structural element.","evidence":"cDNA cloning, sequence analysis, and double immunofluorescence with anti-lamin antibodies in human cells","pmids":["1541630","1541636"],"confidence":"High","gaps":["No functional data; structure-function relationship of domains unknown","No interacting partners identified"]},{"year":1995,"claim":"Immunodepletion and reconstitution experiments demonstrated that NuMA is essential for organizing microtubule asters and that its C-terminal tail mediates microtubule bundling, establishing NuMA as a direct structural organizer of mitotic spindle poles rather than a passive nuclear scaffold.","evidence":"Cell-free mitotic aster assembly, immunodepletion with recombinant protein rescue, EM of purified protein, CDK1 phosphosite mutagenesis","pmids":["7593190","7781599","7769006"],"confidence":"High","gaps":["Motor-dependent transport mechanism to poles not yet defined","Regulation of NuMA–MT interaction beyond CDK1 unknown"]},{"year":1999,"claim":"EM and self-assembly studies revealed that NuMA oligomerizes into multiarm structures through C-terminal interactions and forms an insoluble matrix at spindle poles independent of microtubules, explaining how NuMA provides a structural scaffold that anchors MT minus-ends.","evidence":"In vitro assembly with EM, biochemical fractionation, immunogold EM of spindle poles","pmids":["10075938","10098933"],"confidence":"High","gaps":["Regulation of assembly/disassembly cycle unclear","How the matrix integrates with pericentriolar material not resolved"]},{"year":2000,"claim":"Live imaging and biochemistry established that dynein–dynactin actively transports NuMA to spindle poles after nuclear envelope breakdown, forming a mitosis-specific complex whose disruption blocks pole assembly—linking NuMA's motor-dependent transport to its structural role.","evidence":"Live GFP imaging, immunoprecipitation, gel filtration, dynamitin overexpression in mammalian cells","pmids":["10811826"],"confidence":"High","gaps":["Which NuMA domains contact dynein LIC not yet mapped","Regulation of complex formation/dissociation unclear"]},{"year":2004,"claim":"Discovery that LGN acts as a conformational switch binding both NuMA and Gαi resolved how NuMA reaches the cell cortex for spindle orientation, establishing the LGN/Gαi/NuMA ternary complex as the cortical force-generating pathway, with CDK1-mediated cyclin B degradation releasing NuMA from dynein at anaphase.","evidence":"FRET biosensor, yeast two-hybrid, RNAi, live imaging; Xenopus egg extract with non-degradable cyclin B","pmids":["15537540","14710193"],"confidence":"High","gaps":["Whether LGN and direct MT binding are mutually exclusive in vivo not tested","Cortical force magnitude not measured"]},{"year":2009,"claim":"Conditional NuMA knockout in mice proved NuMA is essential in vivo for tethering spindle microtubules to centrosomes under tension, resolving the question of whether NuMA has a unique function beyond what centrosomes and other crosslinkers provide.","evidence":"Conditional loss-of-function allele in mouse primary cells, live imaging","pmids":["19255246"],"confidence":"High","gaps":["Whether NuMA's tethering and cortical functions are independently essential in vivo not separated","Tissue-specific requirements not fully explored"]},{"year":2011,"claim":"Crystal structures of LGN bound to NuMA versus mInsc revealed mutually exclusive binding to the same TPR surface, establishing that Par3/mInsc/LGN and NuMA/LGN/Gαi act sequentially rather than simultaneously in orienting the spindle.","evidence":"X-ray crystallography with in vitro competition binding assays","pmids":["21816348"],"confidence":"High","gaps":["Temporal handoff mechanism from mInsc to NuMA in vivo not directly visualized","Whether additional factors modulate the switch unknown"]},{"year":2013,"claim":"Multiple groups established that CDK1 phosphorylation of NuMA T2055 suppresses cortical NuMA during metaphase while PP2A counteracts this, and that a C-terminal lipid-binding domain mediates LGN-independent cortical association in anaphase—revealing a phospho-switch coupling mitotic progression to spindle positioning forces.","evidence":"In vitro kinase assays, phosphomutant rescue, lipid-binding assays, live imaging in human cells and keratinocytes","pmids":["23921553","24371089","24109598"],"confidence":"High","gaps":["Precise PIP species specificity not determined","How cortical NuMA domains are spatially restricted to the cell equator at anaphase unclear"]},{"year":2014,"claim":"Direct binding of NuMA to phosphoinositides (PIP/PIP2) was demonstrated, and PIP depletion abolished cortical NuMA, establishing phosphoinositides as the primary cortical receptor for NuMA during anaphase when LGN/Gαi are dispensable.","evidence":"In vitro lipid-binding assay, chemical/enzymatic PIP depletion, immunofluorescence, siRNA rescue","pmids":["24996901"],"confidence":"High","gaps":["Whether PIP2 clustering or local PIP2 generation provides spatial specificity unknown","Structural basis of NuMA–PIP2 interaction not resolved"]},{"year":2016,"claim":"Aurora-A phosphorylation was shown to control NuMA's dynamic exchange between spindle poles and the cortex without affecting MT or LGN binding affinity, and in vivo mutagenesis of NuMA's MT-binding domain in mouse skin caused spindle orientation defects and neonatal lethality, demonstrating NuMA's direct MT engagement is essential for tissue morphogenesis.","evidence":"In vitro kinase assay, FRAP, phosphomutants; conditional MT-binding domain knockout in mouse epidermis","pmids":["26832443","26765568"],"confidence":"High","gaps":["How Aurora-A phosphorylation alters NuMA mobility mechanistically (conformational change?) not defined","Relative contributions of dynein-mediated vs. direct MT-binding functions in different tissues not quantified"]},{"year":2017,"claim":"Quantitative imaging and structural studies revealed that NuMA localizes to MT minus-ends independently of dynein and recruits dynactin there, while a crystal structure showed Importin-α/β masks NuMA's MT-binding region with RanGTP releasing the inhibition—establishing NuMA as a bona fide minus-end cargo adaptor regulated by the Ran gradient.","evidence":"Laser ablation, domain deletion rescue, X-ray crystallography of Importin-α/NuMA complex, in vitro MT-binding assay","pmids":["29185983","28939615"],"confidence":"High","gaps":["Whether Ran gradient regulation operates at the cortex as well as at poles unclear","Minus-end recognition mechanism (structural feature of minus-end) not identified"]},{"year":2018,"claim":"Optogenetic cortical targeting showed that NuMA—not dynein alone—is sufficient to generate spindle-pulling forces by assembling cortical focal clusters of dynein–dynactin; Plk1 was identified as an additional kinase phosphorylating NuMA to tune cortical levels, adding a third kinase input to the cortical regulatory circuit.","evidence":"Light-induced cortical targeting with AID depletion and domain dissection; Plk1 in vitro kinase assay with pharmacological inhibition and FRAP","pmids":["29848445","30456393"],"confidence":"High","gaps":["How CDK1, Aurora-A, and Plk1 phosphorylation events are integrated on the same NuMA molecule not resolved","Stoichiometry of NuMA within cortical clusters unknown"]},{"year":2020,"claim":"Structural determination of NuMA–LIC1/LIC2 interfaces identified two distinct dynein light-intermediate-chain-binding sites (hook domain and CC1-box), both essential for spindle positioning, providing the molecular basis for NuMA's function as a dynein cargo adaptor.","evidence":"X-ray crystallography of NuMA–LIC1/2 interfaces, mutagenesis, functional cell division assay","pmids":["32413290"],"confidence":"High","gaps":["Whether the two LIC-binding sites engage one or two dynein complexes simultaneously unknown","Regulation of LIC-binding site accessibility not addressed"]},{"year":2013,"claim":"Discovery of NuMA's interphase nuclear functions revealed it binds p53 and recruits CDK8/Mediator selectively to p21 but not PUMA promoters, establishing NuMA as a transcriptional cofactor that confers target-gene selectivity to the p53 response.","evidence":"Co-immunoprecipitation, ChIP, siRNA knockdown with selective gene expression readout","pmids":["23589328"],"confidence":"Medium","gaps":["Structural basis for selective promoter targeting unknown","Whether NuMA acts on other transcription factors besides p53 not tested","Independent replication needed"]},{"year":2014,"claim":"NuMA was found to accumulate at DNA damage sites in a PARylation-dependent manner, interact with SNF2h to promote chromatin decompaction and homologous recombination, linking NuMA's nuclear matrix role to active DNA repair.","evidence":"Co-IP, FRAP, laser microirradiation, siRNA with chromosomal DSB repair assays","pmids":["24753406"],"confidence":"Medium","gaps":["Whether NuMA is directly PARylated at breaks or recruited via PAR binding unclear","Relationship to NuMA's interaction with tankyrase-1 at spindle poles not addressed"]},{"year":2019,"claim":"NuMA was shown to sequester 53BP1 in undamaged nuclei and limit its access to breaks, with NuMA depletion altering PARP inhibitor sensitivity in BRCA1-null cells, revealing NuMA as a regulator of DNA repair pathway choice.","evidence":"Co-IP, FRAP, NHEJ assay, class-switch recombination assay, PARP inhibitor sensitivity","pmids":["30812030"],"confidence":"Medium","gaps":["Mechanism by which NuMA restrains 53BP1 mobility not structurally defined","In vivo relevance for tumor suppression not tested"]},{"year":2021,"claim":"NuMA was shown to undergo liquid–liquid phase separation at mitotic entry, regulated by Aurora-A, concentrating tubulins and the MT depolymerase Kif2A at poles to promote poleward flux—providing a biophysical framework for how NuMA's self-assembly creates a functional spindle pole compartment.","evidence":"In vitro LLPS reconstitution, live cell condensate imaging, phosphomimetic/phosphodead mutants, siRNA","pmids":["34887424"],"confidence":"Medium","gaps":["Whether LLPS and the previously described insoluble matrix represent the same or distinct states unclear","Contribution of LLPS versus oligomeric cross-linking to pole integrity not separated","Independent replication of LLPS findings needed"]},{"year":null,"claim":"Key unresolved questions include how the three mitotic kinase inputs (CDK1, Aurora-A, Plk1) are integrated on a single NuMA molecule to produce graded cortical versus polar localization, the structural basis of NuMA's phosphoinositide binding and minus-end recognition, and the extent to which NuMA's interphase nuclear functions (p53 cofactor, rDNA transcription, DNA repair pathway choice) are mechanistically independent or coupled through a common scaffolding principle.","evidence":"","pmids":[],"confidence":"High","gaps":["Integrated phospho-regulation model lacking","No structural data for NuMA–PIP2 or NuMA–minus-end interfaces","Whether nuclear and mitotic NuMA pools are regulated by shared or distinct mechanisms unknown"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[0,3,7,22,29,30]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[20,32,19]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[23,31,22]},{"term_id":"GO:0008289","term_label":"lipid binding","supporting_discovery_ids":[15,16]},{"term_id":"GO:0140110","term_label":"transcription regulator activity","supporting_discovery_ids":[37,42]}],"localization":[{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[2,46,37,42]},{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,1,3,19,32]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,22,23,29]},{"term_id":"GO:0005886","term_label":"plasma membrane","supporting_discovery_ids":[6,14,15,16,23]},{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[42]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1,3,4,14,19,22,23]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[36,38]},{"term_id":"R-HSA-74160","term_label":"Gene expression (Transcription)","supporting_discovery_ids":[37,42]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[6,25,26]}],"complexes":["NuMA–dynein–dynactin","LGN/Gαi/NuMA","NuMA–tankyrase-1"],"partners":["DYNC1H1","DCTN1","GPSM2","GNAI1","AURKA","EPB41","TNKS","TP53"],"other_free_text":[]},"mechanistic_narrative":"NuMA is a large coiled-coil protein that functions as a master organizer of mitotic spindle architecture and a regulator of nuclear processes during interphase. During mitosis, CDK1 phosphorylation triggers NuMA's release from the nucleus; NuMA is then transported poleward by cytoplasmic dynein–dynactin, where it self-assembles into an insoluble cross-linking matrix that focuses microtubule minus-ends at spindle poles—a function essential for centrosome–spindle attachment under tension and for acentrosomal spindle bipolarization [PMID:8898198, PMID:19255246, PMID:31782546]. At the cell cortex, NuMA is recruited by LGN/Gαi and directly by phosphoinositides to cluster dynein–dynactin force generators that pull on astral microtubules for spindle orientation, with cortical NuMA levels dynamically tuned by CDK1, Aurora-A, and Plk1 phosphorylation and opposed by PP2A dephosphorylation [PMID:15537540, PMID:24996901, PMID:23921553, PMID:26832443]. In interphase, NuMA resides in the nuclear matrix where it contributes to p53 target-gene selectivity by recruiting CDK8/Mediator, facilitates rDNA transcription through RNA Pol I association, and regulates DNA double-strand break repair by controlling SNF2h-mediated chromatin remodeling and 53BP1 dynamics [PMID:23589328, PMID:28981686, PMID:24753406, PMID:30812030]."},"prefetch_data":{"uniprot":{"accession":"Q14980","full_name":"Nuclear mitotic apparatus protein 1","aliases":["Nuclear matrix protein-22","NMP-22","Nuclear mitotic apparatus protein","NuMA protein","SP-H antigen"],"length_aa":2115,"mass_kda":238.3,"function":"Microtubule (MT)-binding protein that plays a role in the formation and maintenance of the spindle poles and the alignement and the segregation of chromosomes during mitotic cell division (PubMed:17172455, PubMed:19255246, PubMed:24996901, PubMed:26195665, PubMed:27462074, PubMed:7769006). Functions to tether the minus ends of MTs at the spindle poles, which is critical for the establishment and maintenance of the spindle poles (PubMed:11956313, PubMed:12445386). Plays a role in the establishment of the mitotic spindle orientation during metaphase and elongation during anaphase in a dynein-dynactin-dependent manner (PubMed:23870127, PubMed:24109598, PubMed:24996901, PubMed:26765568). In metaphase, part of a ternary complex composed of GPSM2 and G(i) alpha proteins, that regulates the recruitment and anchorage of the dynein-dynactin complex in the mitotic cell cortex regions situated above the two spindle poles, and hence regulates the correct oritentation of the mitotic spindle (PubMed:22327364, PubMed:23027904, PubMed:23921553). During anaphase, mediates the recruitment and accumulation of the dynein-dynactin complex at the cell membrane of the polar cortical region through direct association with phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2), and hence participates in the regulation of the spindle elongation and chromosome segregation (PubMed:22327364, PubMed:23921553, PubMed:24371089, PubMed:24996901). Also binds to other polyanionic phosphoinositides, such as phosphatidylinositol 3-phosphate (PIP), lysophosphatidic acid (LPA) and phosphatidylinositol triphosphate (PIP3), in vitro (PubMed:24371089, PubMed:24996901). Also required for proper orientation of the mitotic spindle during asymmetric cell divisions (PubMed:21816348). Plays a role in mitotic MT aster assembly (PubMed:11163243, PubMed:11229403, PubMed:12445386). Involved in anastral spindle assembly (PubMed:25657325). Positively regulates TNKS protein localization to spindle poles in mitosis (PubMed:16076287). Highly abundant component of the nuclear matrix where it may serve a non-mitotic structural role, occupies the majority of the nuclear volume (PubMed:10075938). Required for epidermal differentiation and hair follicle morphogenesis (By similarity)","subcellular_location":"Cytoplasm, cytosol; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome; Cytoplasm, cytoskeleton, spindle pole","url":"https://www.uniprot.org/uniprotkb/Q14980/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/NUMA1","classification":"Common Essential","n_dependent_lines":700,"n_total_lines":1208,"dependency_fraction":0.5794701986754967},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000137497","cell_line_id":"CID001100","localizations":[{"compartment":"nucleoplasm","grade":3},{"compartment":"chromatin","grade":2}],"interactors":[{"gene":"H1F0","stoichiometry":10.0},{"gene":"NUCKS1","stoichiometry":10.0},{"gene":"HIST1H1D","stoichiometry":10.0},{"gene":"H1FX","stoichiometry":10.0},{"gene":"HIST1H2AC","stoichiometry":10.0},{"gene":"H2AFY2","stoichiometry":10.0},{"gene":"HIST2H2AB","stoichiometry":10.0},{"gene":"HIST3H2BB","stoichiometry":10.0},{"gene":"HMGN5","stoichiometry":4.0},{"gene":"MYO1E","stoichiometry":4.0}],"url":"https://opencell.sf.czbiohub.org/target/CID001100","total_profiled":1310},"omim":[{"mim_id":"616919","title":"FERM AND PDZ DOMAINS-CONTAINING PROTEIN 1; FRMPD1","url":"https://www.omim.org/entry/616919"},{"mim_id":"613529","title":"CENTROSOMAL PROTEIN, 152-KD; CEP152","url":"https://www.omim.org/entry/613529"},{"mim_id":"613004","title":"HUNTINGTIN; HTT","url":"https://www.omim.org/entry/613004"},{"mim_id":"612376","title":"ACUTE PROMYELOCYTIC LEUKEMIA; APL","url":"https://www.omim.org/entry/612376"},{"mim_id":"604260","title":"SIGNAL TRANSDUCER AND ACTIVATOR OF TRANSCRIPTION 5B; STAT5B","url":"https://www.omim.org/entry/604260"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Enhanced","locations":[{"location":"Nucleoplasm","reliability":"Enhanced"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NUMA1"},"hgnc":{"alias_symbol":["NUMA"],"prev_symbol":[]},"alphafold":{"accession":"Q14980","domains":[{"cath_id":"1.10.418.10","chopping":"6-152","consensus_level":"high","plddt":72.8283,"start":6,"end":152},{"cath_id":"-","chopping":"497-533_566-824","consensus_level":"medium","plddt":76.0592,"start":497,"end":824},{"cath_id":"1.20.5","chopping":"826-887","consensus_level":"medium","plddt":71.3695,"start":826,"end":887},{"cath_id":"1.10.287","chopping":"1036-1101_1122-1327","consensus_level":"medium","plddt":76.9103,"start":1036,"end":1327},{"cath_id":"1.20.5","chopping":"1423-1559","consensus_level":"high","plddt":78.7312,"start":1423,"end":1559}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q14980","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q14980-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q14980-F1-predicted_aligned_error_v6.png","plddt_mean":64.62},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NUMA1","jax_strain_url":"https://www.jax.org/strain/search?query=NUMA1"},"sequence":{"accession":"Q14980","fasta_url":"https://rest.uniprot.org/uniprotkb/Q14980.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q14980/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q14980"}},"corpus_meta":[{"pmid":"8898198","id":"PMC_8898198","title":"A 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NuMA contains this RXXPDG motif and is a novel tankyrase partner.","date":"2002","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/12080061","citation_count":131,"is_preprint":false},{"pmid":"16076287","id":"PMC_16076287","title":"NuMA is a major acceptor of poly(ADP-ribosyl)ation by tankyrase 1 in mitosis.","date":"2005","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/16076287","citation_count":131,"is_preprint":false},{"pmid":"12445386","id":"PMC_12445386","title":"LGN blocks the ability of NuMA to bind and stabilize microtubules. A mechanism for mitotic spindle assembly regulation.","date":"2002","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/12445386","citation_count":126,"is_preprint":false},{"pmid":"21444683","id":"PMC_21444683","title":"A lateral belt of cortical LGN and NuMA guides mitotic spindle movements and planar division in neuroepithelial cells.","date":"2011","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/21444683","citation_count":123,"is_preprint":false},{"pmid":"19255246","id":"PMC_19255246","title":"Requirements for NuMA in maintenance and establishment of mammalian spindle poles.","date":"2009","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/19255246","citation_count":119,"is_preprint":false},{"pmid":"10189366","id":"PMC_10189366","title":"A nonerythroid isoform of protein 4.1R interacts with the nuclear mitotic apparatus (NuMA) protein.","date":"1999","source":"The Journal of cell 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abolishes normal spindle pole assembly, producing chromatin-associated irregular microtubule arrays; a NuMA tail subdomain induces microtubule aster formation by mediating microtubule bundling.\",\n      \"method\": \"Immunodepletion from Xenopus egg extracts, Co-immunoprecipitation, in vitro aster assembly assay\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstitution in egg extracts with immunodepletion and rescue, replicated by multiple subsequent studies\",\n      \"pmids\": [\"8898198\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"NuMA transport to spindle poles upon nuclear envelope breakdown is powered by cytoplasmic dynein and dynactin along microtubule minus ends; large cytoplasmic NuMA aggregates stream poleward in association with dynactin (Arp1) and dynein; immunoprecipitation and gel filtration demonstrate a reversible, mitosis-specific NuMA–dynein–dynactin complex; disruption of dynactin or dynein blocks NuMA translocation and spindle pole assembly.\",\n      \"method\": \"Live GFP imaging, immunoprecipitation, gel filtration, dynamitin overexpression, dynein antibody inhibition\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — multiple orthogonal methods in one study, consistent with prior immunodepletion data\",\n      \"pmids\": [\"10811826\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1992,\n      \"finding\": \"NuMA is a 236–238 kDa nuclear protein with a long central coiled-coil domain (~1,485 aa) flanked by globular N- and C-terminal domains; it dissociates from condensing chromosomes early in prophase before lamina disintegration and reassociates with telophase chromosomes before lamin accumulation, indicating a role in nuclear reformation.\",\n      \"method\": \"cDNA cloning, sequence analysis, double immunofluorescence with anti-NuMA and anti-lamin antibodies\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — primary structure determination with domain mapping, independently confirmed by Compton et al. 1992\",\n      \"pmids\": [\"1541630\", \"1541636\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"NuMA is required for organizing microtubules into aster-like mitotic arrays; immunodepletion of NuMA from mitotic cell extracts prevents aster formation, and addition of purified recombinant NuMA fully rescues aster assembly; NuMA is phosphorylated upon aster assembly and is only required in late stages of aster formation.\",\n      \"method\": \"Cell-free mitotic aster assembly assay, immunodepletion, recombinant protein rescue\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstitution with immunodepletion and rescue by purified protein\",\n      \"pmids\": [\"7593190\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"Mutation of the predicted CDK1 (p34cdc2) phosphorylation site at Thr2040 of NuMA (alone or combined with other site mutations) abolishes NuMA's ability to associate with spindle microtubules; instead the mutant concentrates at the plasma membrane, causing disorganized spindles, cytokinesis failure, and micronucleation. This demonstrates that mitosis-specific phosphorylation controls NuMA's spindle interaction.\",\n      \"method\": \"Site-directed mutagenesis, transient expression, immunofluorescence, flow cytometry\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — mutagenesis with defined phosphorylation site linked to specific phenotype\",\n      \"pmids\": [\"7769006\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1993,\n      \"finding\": \"Expression of NuMA lacking its globular head domain results in cytokinesis failure and micronuclei formation; expression of NuMA lacking the globular tail domain prevents nuclear targeting and spindle binding, causing micronucleation in daughter cells. Wild-type NuMA overexpression rescues nuclear assembly defects in RCC1 temperature-sensitive cells. NuMA function is required for terminal phases of chromosome separation and/or nuclear reassembly.\",\n      \"method\": \"Dominant-negative expression, temperature-sensitive mutant cell rescue, immunofluorescence\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — domain deletion mutants with well-defined phenotypes, rescue experiment\",\n      \"pmids\": [\"8432734\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"LGN (mammalian Pins homolog) directly binds NuMA through its N-terminal domain and recruits NuMA to the cell cortex during mitosis; LGN behaves as a conformational switch—its closed state has N- and C-termini interacting, but NuMA or Gαi can open the switch, allowing simultaneous binding to both, resulting in their cortical localization. LGN–NuMA interaction is required for spindle oscillation.\",\n      \"method\": \"FRET biosensor, yeast two-hybrid, overexpression, RNAi, live imaging\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — FRET structural insight plus functional rescue, multiple orthogonal approaches\",\n      \"pmids\": [\"15537540\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2001,\n      \"finding\": \"LGN (human Pins-related protein) binds the C-terminal tail of NuMA; LGN blocks NuMA-dependent stabilization and bundling of microtubules. In vitro assays show NuMA binds MTs directly, and the MT-binding domain overlaps by 10 aa with the LGN-binding domain, indicating steric exclusion explains LGN's inhibitory effect on NuMA spindle function.\",\n      \"method\": \"In vitro MT binding/stabilization assays, Xenopus egg extract aster assay, domain mapping\",\n      \"journal\": \"Nature cell biology / Current biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro binding and MT stabilization assays with domain mutagenesis, two papers\",\n      \"pmids\": [\"11781568\", \"12445386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"A 135 kDa nonerythroid isoform of protein 4.1R directly interacts with NuMA; minimal binding involves 4.1R exons 20–21 and NuMA residues 1788–1810; 4.1R and NuMA co-localize in interphase nuclei and redistribute to spindle poles, where 4.1R is part of a NuMA–dynein–dynactin complex during mitosis.\",\n      \"method\": \"Yeast two-hybrid, in vitro binding assay, co-immunoprecipitation, co-immunolocalization\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal binding methods with domain mapping, single lab\",\n      \"pmids\": [\"10189366\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"NuMA contains an RXXPDG motif that mediates direct binding to the ankyrin-repeat domain of tankyrase-1 and tankyrase-2 (poly-ADP-ribose polymerases); this interaction contributes to the known co-localization of tankyrase and NuMA at mitotic spindle poles.\",\n      \"method\": \"Yeast two-hybrid, in vitro binding, co-localization\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — yeast two-hybrid plus in vitro binding, single lab\",\n      \"pmids\": [\"12080061\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"NuMA is a major acceptor of poly(ADP-ribosyl)ation (PARsylation) by tankyrase 1 in mitosis; tankyrase 1 and NuMA association increases at mitotic onset concomitant with NuMA PARsylation; siRNA knockdown of tankyrase 1 eliminates NuMA PARsylation; conversely, NuMA knockdown causes complete loss of tankyrase 1 from spindle poles.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, siRNA knockdown\",\n      \"journal\": \"The Biochemical journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP with siRNA rescue, single lab, single paper\",\n      \"pmids\": [\"16076287\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"Poly(ADP-ribose) (pADPr) made by PARP-5a/tankyrase-1 localizes to spindle poles; pADPr-coated beads trigger microtubule aster assembly in mitotic HeLa lysate; the rod domain of NuMA binds directly to pADPr chains, suggesting pADPr provides dynamic cross-linking at spindle poles by binding NuMA.\",\n      \"method\": \"In vitro pADPr bead aster assay, recombinant domain binding, immuno-EM\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro aster reconstitution plus direct binding, single lab\",\n      \"pmids\": [\"19759176\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"NuMA is phosphorylated by CDK1 (Cdc2 kinase) in a mitosis-specific manner as shown in vivo with 32P-labeling; phosphorylation shifts NuMA to a higher apparent molecular weight form; this modification coincides with NuMA's release from the nucleus and redistribution to the spindle.\",\n      \"method\": \"32P metabolic labeling, phosphatase treatment, immunoblot in synchronized cells\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — biochemical characterization in synchronized cells with phosphatase controls, single lab\",\n      \"pmids\": [\"9202389\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"Cyclin B degradation at anaphase onset leads to NuMA dephosphorylation and its release from dynein, dynactin, and spindle poles; a non-degradable cyclin B (Δ90) keeps NuMA phosphorylated and stably associated with dynein/dynactin at persistent spindle poles that fail to disassemble.\",\n      \"method\": \"Xenopus egg extract, non-degradable cyclin B expression, co-immunoprecipitation, immunofluorescence\",\n      \"journal\": \"EMBO reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — biochemical rescue experiment with defined cyclin B mutant, single lab\",\n      \"pmids\": [\"14710193\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"CDK1 phosphorylates NuMA at T2055 to negatively regulate its cortical localization during metaphase; PPP2CA (PP2A) phosphatase counteracts this; CDK1 inactivation in anaphase increases dephosphorylated cortical NuMA, enriching cortical dynein and driving spindle elongation. Thus, NuMA phosphorylation status by CDK1 couples mitotic progression with spindle behavior.\",\n      \"method\": \"In vitro kinase assay, phosphorylation-site mutagenesis, immunofluorescence, siRNA knockdown in human cells\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct kinase assay with mutagenesis, functional rescue, single lab with multiple orthogonal methods\",\n      \"pmids\": [\"23921553\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"A lipid/membrane-binding domain at the C-terminus of NuMA mediates LGN/Gαi-independent cortical association during anaphase; this binding is inhibited by CDK1 phosphorylation during prophase and metaphase and activated upon CDK1 inactivation at anaphase; replacing endogenous NuMA with membrane-binding-deficient NuMA reduces anaphase cortical dynein and impairs chromosome separation.\",\n      \"method\": \"Domain mapping, phosphorylation-site mutagenesis, live cell imaging, siRNA rescue experiments, lipid-binding assays\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — domain-function dissection with phosphomutants and rescue, multiple methods\",\n      \"pmids\": [\"24371089\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"NuMA directly associates with phosphoinositides PtdInsP (PIP) and PtdInsP2 (PIP2) in vitro; depletion of PIP/PIP2 by chemical or enzymatic means prevents NuMA cortical localization during mitosis; increasing PIP2 augments cortical NuMA. During anaphase, LGN/Gαi are dispensable for NuMA cortical enrichment but PIP/PIP2 are required.\",\n      \"method\": \"In vitro lipid-binding assay, chemical/enzymatic PIP depletion, immunofluorescence, siRNA knockdown\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct biochemical binding plus functional depletion with multiple rescue strategies\",\n      \"pmids\": [\"24996901\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"Aurora-A directly phosphorylates the C-terminus of NuMA on three serine residues, including Ser1969; this phosphorylation governs the dynamic exchange of NuMA between cytoplasm/spindle poles and cortex; partial Aurora-A inhibition traps NuMA at spindle poles and prevents cortical enrichment. Aurora-A phosphorylation of NuMA does not affect LGN or MT binding affinity but controls NuMA mobility at poles.\",\n      \"method\": \"In vitro kinase assay, phospho-site mutagenesis, FRAP, immunofluorescence, kinase inhibition in human cells\",\n      \"journal\": \"Current biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro kinase assay with mutagenesis, FRAP, and functional phenotype rescue\",\n      \"pmids\": [\"26832443\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Plk1 directly interacts with and phosphorylates NuMA; acute Plk1 inactivation enriches cortical NuMA/LGN/dynein during metaphase; Plk1-mediated NuMA phosphorylation controls NuMA cortical localization dynamics and spindle orientation.\",\n      \"method\": \"In vitro kinase assay, co-immunoprecipitation, pharmacological Plk1 inhibition, FRAP, immunofluorescence\",\n      \"journal\": \"Life science alliance\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1–2 — in vitro kinase assay plus functional readout, single lab\",\n      \"pmids\": [\"30456393\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"NuMA is an essential tether linking bulk spindle microtubules to centrosomes; conditional loss of NuMA function in mice shows that centrosomes provide initial spindle focusing but centrosome-spindle attachment under tension fails without NuMA, and kinetochore fiber maintenance at poles is lost; without both centrosomes and NuMA, all spindle focusing fails.\",\n      \"method\": \"Conditional loss-of-function allele (mouse genetics), primary cell culture, immunofluorescence, live imaging\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean conditional KO with defined cellular and structural phenotypes, in vivo\",\n      \"pmids\": [\"19255246\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"NuMA self-assembles in vitro into multiarm oligomers (up to 10–12 arms) through C-terminal globular domain interactions; each arm corresponds to a NuMA dimer; electron microscopy and computer modeling show these 12-arm oligomers are the structural units of a quasi-hexagonal nuclear scaffold observed upon NuMA overexpression in HeLa cells.\",\n      \"method\": \"In vitro assembly, electron microscopy, computer modeling, overexpression in HeLa cells\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution of NuMA oligomers with EM structural validation\",\n      \"pmids\": [\"10075938\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1995,\n      \"finding\": \"The NuMA rod domain forms a 207 nm long parallel, in-register double-stranded coiled-coil (the longest known); electron microscopy of purified recombinant fragments and full-length protein confirms tripartite structure with flanking globular domains; the SPN-3 antibody epitope maps to residues 255–267 in the first helical subdomain.\",\n      \"method\": \"Electron microscopy of recombinant protein, chemical cross-linking, circular dichroism, epitope mapping\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — direct structural characterization of purified protein by EM and biophysical methods\",\n      \"pmids\": [\"7781599\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NuMA recruits dynactin to microtubule minus-ends; NuMA localizes to new minus-ends independently of dynein and faster than dynactin; a C-terminal region outside NuMA's canonical MT-binding domain mediates minus-end localization independently of γ-TuRC, CAMSAP1, or KANSL1/3; both NuMA's minus-end-binding and dynein-dynactin-binding modules are required to rescue bipolar spindle organization.\",\n      \"method\": \"Quantitative live imaging, laser ablation, domain deletion mutants, siRNA knockdown with rescue\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — quantitative imaging with laser ablation and domain-specific rescue, multiple orthogonal approaches\",\n      \"pmids\": [\"29185983\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"Cortical targeting of NuMA (but not dynein alone) is sufficient to generate spindle-pulling forces; NuMA assembles specialized focal structures at the cortex that cluster multiple dynein-dynactin force-generating modules; NuMA's N-terminal long arm (dynein-dynactin recruitment), dynein-based MT gliding, and direct MT-binding activity are all required; NuMA clustering is essential for spindle positioning but not spindle-pole focusing.\",\n      \"method\": \"Light-induced cortical targeting (optogenetics/reconstitution), AID-based depletion, live imaging, domain dissection\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — light-induced reconstitution system with domain dissection, multiple orthogonal methods\",\n      \"pmids\": [\"29848445\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Crystal structures of LGN/NuMA and LGN/mInsc complexes reveal that NuMA and Inscuteable (mInsc) interact with the same LGN TPR domain in a mutually exclusive manner; mInsc binds with higher affinity; biochemical competition assays confirm this exclusivity, suggesting Par3/mInsc/LGN and NuMA/LGN/Gαi complexes play sequential rather than simultaneous roles.\",\n      \"method\": \"X-ray crystallography, in vitro competition binding assays, cell biology studies\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structures of both complexes with orthogonal biochemical validation\",\n      \"pmids\": [\"21816348\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"Ric-8A, a Gαi guanine nucleotide exchange factor, catalytically dissociates Gαi-GDP/LGN/NuMA complexes in vitro by releasing activated Gαi-GTP, which concomitantly liberates NuMA from LGN; this mechanism may regulate microtubule pulling forces on centrosomes during cell division.\",\n      \"method\": \"In vitro GEF assay with purified components, biochemical complex dissociation assay\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution assay with purified proteins, single lab\",\n      \"pmids\": [\"16275912\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"Ric-8A and Gαi recruit LGN, NuMA, and dynein to the cell cortex during mitosis to orient the mitotic spindle; pertussis toxin (blocking Ric-8A GEF activity for Gαi), Ric-8A siRNA, or Gαi siRNA each impair cortical LGN, NuMA, and dynein localization and disturb integrin-dependent spindle orientation.\",\n      \"method\": \"siRNA knockdown, pertussis toxin treatment, live GFP-tubulin imaging, immunofluorescence\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple loss-of-function approaches with defined localization and orientation phenotype, single lab\",\n      \"pmids\": [\"20479129\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CYLD deubiquitinase deubiquitinates the cortical polarity protein dishevelled, enhancing dishevelled–NuMA interaction and promoting cortical localization of NuMA and the dynein/dynactin complex; CYLD also stabilizes astral microtubules to regulate spindle orientation.\",\n      \"method\": \"Co-immunoprecipitation, ubiquitination assay, siRNA knockdown, immunofluorescence\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — biochemical deubiquitination assay plus co-IP and loss-of-function, single lab\",\n      \"pmids\": [\"24469800\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"NuMA's 4.1-binding domain is required for stabilizing NuMA at the cell cortex of keratinocytes (shown by FRAP); loss of 4.1/NuMA interaction causes spindle orientation defects; NuMA is required to recruit dynactin to the cortex; at anaphase, a CDK1-independent, LGN/4.1-independent mechanism also drives cortical NuMA accumulation.\",\n      \"method\": \"FRAP, domain mutant expression, siRNA knockdown, immunofluorescence in primary keratinocytes\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — FRAP with domain mutants and functional spindle orientation readout, single lab\",\n      \"pmids\": [\"24109598\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"NuMA's direct MT-binding domain (targeting MT tips) is essential for spindle orientation establishment in keratinocytes; dynein/dynactin alone is insufficient; loss of NuMA–MT interactions in mouse skin causes spindle orientation defects, epidermal differentiation failure, and neonatal lethality; in adult mice, loss of NuMA-MT interactions disrupts hair follicle morphogenesis.\",\n      \"method\": \"MT-binding domain mutagenesis, conditional knockout in mouse skin, live imaging, immunofluorescence\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — in vivo conditional KO with domain-specific mutagenesis, defined in vivo phenotype\",\n      \"pmids\": [\"26765568\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Crystal structure of Importin-α bound to the NuMA C-terminus reveals a novel NLS binding pattern enabling selective NLS recognition; Importin-β, in the presence of Importin-α, sterically masks a high-affinity MT-binding region of NuMA C-terminal to the NLS, thereby inhibiting NuMA's MT-bundling activity; RanGTP releases this inhibition.\",\n      \"method\": \"X-ray crystallography, in vitro MT-binding assay, Importin-β binding assay, mutagenesis\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure with functional in vitro validation by MT-binding assay and mutagenesis\",\n      \"pmids\": [\"28939615\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"NuMA contains two dynein LIC-binding sites: a hook domain contacting LIC1/LIC2 through a conserved hydrophobic patch (shared with Hook adaptors) and a CC1-box-like motif within its coiled-coil; both LIC-binding sites are essential for correct spindle positioning and cell division.\",\n      \"method\": \"X-ray crystallography of NuMA–LIC1/2 interfaces, mutagenesis, functional cell division assay\",\n      \"journal\": \"Structure\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — crystal structure with mutagenesis and functional validation\",\n      \"pmids\": [\"32413290\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1999,\n      \"finding\": \"Once NuMA is incorporated into the insoluble spindle pole matrix in vivo or in vitro, its insolubility becomes independent of microtubules; immunogold EM shows NuMA at an electron-dense material distinct from pericentriolar material that appears to anchor microtubule ends; NuMA is essential for formation of this insoluble matrix.\",\n      \"method\": \"Immunogold electron microscopy, biochemical fractionation, immunodepletion of cell-free extracts\",\n      \"journal\": \"Cell motility and the cytoskeleton\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — EM localization with biochemical fractionation and immunodepletion, single lab\",\n      \"pmids\": [\"10098933\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"Rae1 (mRNA export factor) has a mitosis-specific interaction with NuMA; Rae1 binds a specific site on NuMA that would convert a NuMA dimer to a tetravalent MT crosslinker; reducing Rae1 or increasing NuMA disrupts spindle architecture; co-depletion or co-overexpression of Rae1 with NuMA prevents aberrant spindle formation.\",\n      \"method\": \"Co-immunoprecipitation, domain mapping, overexpression, siRNA knockdown in HeLa cells\",\n      \"journal\": \"PNAS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP with domain mapping and epistasis by double perturbation, single lab\",\n      \"pmids\": [\"17172455\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2004,\n      \"finding\": \"NuMA dynamics at spindle poles involve continuous exchange between soluble and spindle-associated pools (t½ ~3 min by FRAP); this exchange requires cellular energy; LGN binding and protein kinase activity both regulate NuMA's dynamic exchange rate on spindle pole asters in cell-free extracts.\",\n      \"method\": \"FRAP in live cells, cell-free aster exchange assay, LGN addition, kinase inhibitors\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — FRAP plus cell-free assay with pharmacological and protein perturbations, single lab\",\n      \"pmids\": [\"15561764\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NuMA undergoes liquid-liquid phase separation (LLPS) at mitotic entry; this is regulated by Aurora-A phosphorylation; KifC1 facilitates NuMA condensate concentration at spindle poles; phase separation is mediated by NuMA's C-terminus and dynein-dynactin binding motif; phase-separated NuMA concentrates tubulins, binds MTs, and enriches Kif2A (a spindle MT depolymerizer) at poles to promote poleward MT flux.\",\n      \"method\": \"In vitro phase separation assay, live cell imaging of condensates, siRNA knockdown, phosphomimetic/phosphodead mutants\",\n      \"journal\": \"Nature communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — in vitro LLPS plus live cell assays, single lab, novel mechanism\",\n      \"pmids\": [\"34887424\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"NuMA accumulates at DNA damage sites in a poly-ADP-ribosylation (PARylation)-dependent manner; NuMA co-immunoprecipitates with the chromatin remodeler SNF2h/SMARCA5 and regulates SNF2h diffusion and its accumulation at DNA breaks; NuMA knockdown impairs chromatin decompaction after DNA cleavage, reduces homologous recombination repair factor recruitment, and impairs DSB repair in chromosomal (not episomal) contexts.\",\n      \"method\": \"Co-immunoprecipitation, FRAP, siRNA knockdown, laser microirradiation, DNA repair assays\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods establishing novel NuMA function in DNA repair, single lab\",\n      \"pmids\": [\"24753406\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"NuMA binds p53 and is required for CDK8 (Mediator complex component) recruitment to p53 target gene promoters; acute NuMA knockdown selectively attenuates p21 induction after DNA damage (impairing cell cycle arrest) but does not affect PUMA induction, demonstrating NuMA confers selectivity to p53-mediated transcription.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, chromatin immunoprecipitation, transcriptional reporter assays\",\n      \"journal\": \"Molecular and cellular biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP with ChIP and selective gene expression readout, single lab\",\n      \"pmids\": [\"23589328\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NuMA interacts with 53BP1 and controls 53BP1 diffusion throughout the nucleoplasm; this interaction is reduced after DNA damage; NuMA prevents 53BP1 accumulation at DNA breaks in the undamaged state; manipulating NuMA expression alters PARP inhibitor sensitivity in BRCA1-null cells and affects end-joining activity and immunoglobulin class switching.\",\n      \"method\": \"Co-immunoprecipitation, FRAP, siRNA knockdown, NHEJ assay, class-switch recombination assay\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal methods for NuMA–53BP1 interaction and functional readout, single lab\",\n      \"pmids\": [\"30812030\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In neurons, NuMA1 is transiently located at the axon initial segment (AIS) where it interacts with scaffolding protein 4.1B and the dynein regulator Lis1; NuMA1 inhibits endocytosis of AIS protein NF186 by impeding Lis1's interaction with doublecortin; silencing NuMA1 or 4.1B disrupts AIS assembly but not maintenance.\",\n      \"method\": \"Differential proteomics, co-immunoprecipitation, shRNA knockdown, endocytosis assay, immunofluorescence in mouse neurons\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP with mechanistic endocytosis assay and loss-of-function phenotype, single lab\",\n      \"pmids\": [\"31727776\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NUMA1 is downregulated in Huntington's disease neural progenitors by miR-124; suppression of NUMA1 in wild-type cells recapitulates HD microtubule bundling defects within axonal growth cones and impairs axon growth; restoring NUMA1 levels (via antagomiR-124) or stabilizing microtubules with epothilone B rescues microtubule organization and axonal growth.\",\n      \"method\": \"Proteomics of growth cones, miRNA manipulation, shRNA knockdown, rescue by antagomiR or drug, live imaging of growth cones\",\n      \"journal\": \"Neuron\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — loss-of-function and rescue experiments with defined microtubule and axon growth phenotype, single lab\",\n      \"pmids\": [\"34793694\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"GAS41 (a protein amplified in gliomas) directly binds the C-terminal rod region of NuMA (Kd ~2×10⁻⁷ M) as shown by dot overlay and surface plasmon resonance; GAS41 is nucleolar in interphase and becomes diffuse in mitosis.\",\n      \"method\": \"Yeast two-hybrid, dot overlay, surface plasmon resonance\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — in vitro binding without functional consequence established, single lab\",\n      \"pmids\": [\"10913114\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"NuMA controls rDNA transcription and mediates nucleolar stress response in a p53-independent manner; NuMA co-immunoprecipitates with RNA Pol I, ribosomal proteins RPL26/RPL24, and B-WICH complex components; NuMA binds 18S and 28S rRNAs and localizes to rDNA promoter regions; NuMA knockdown decreases nascent pre-rRNA synthesis and triggers p27kip1 upregulation.\",\n      \"method\": \"Co-immunoprecipitation, RNA-binding assay, ChIP, siRNA knockdown, nascent RNA labeling\",\n      \"journal\": \"Nucleic acids research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple binding and functional assays establishing novel nucleolar function, single lab\",\n      \"pmids\": [\"28981686\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"NuMA directly binds Astrin (C-terminal tail of NuMA to C-terminus of Astrin); NuMA is required for Astrin recruitment to the mitotic spindle; LGN N-terminus (which blocks NuMA MT binding) also displaces Astrin from NuMA; dynein-mediated transport is required for spindle pole accumulation of Astrin; conversely, reduced Astrin impairs NuMA concentration at spindle poles.\",\n      \"method\": \"Co-immunoprecipitation, GST pulldown, siRNA knockdown, immunofluorescence\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct binding assay plus reciprocal knockdown phenotypes, single lab\",\n      \"pmids\": [\"27462074\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1997,\n      \"finding\": \"NuMA is cleaved during apoptosis; the cleavage site lies between residues 1701 and 1725 generating a stable ~180–200 kDa fragment; cleavage is retarded by TPCK but not by ICE inhibitors or other protease inhibitors tested.\",\n      \"method\": \"Immunoblot, protease inhibitor treatment, apoptosis induction in cell lines\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — cleavage site mapped with pharmacological dissection, single lab\",\n      \"pmids\": [\"9184071\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2003,\n      \"finding\": \"NuMA is preferentially cleaved by caspase-3 during Fas-mediated apoptosis; NuMA cleavage coincides with lamin B and PARP-1 cleavage; NuMA redistribution during apoptosis involves condensation, central nuclear concentration, and encirclement of apoptotic body fragments; NuMA was not cleaved in caspase-3-null MCF-7 cells treated with staurosporine.\",\n      \"method\": \"Immunofluorescence, immunoblot, caspase inhibitors, caspase-3-null cell line, Fas stimulation\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic null cell line plus pharmacological inhibitors, single lab\",\n      \"pmids\": [\"12508117\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 1994,\n      \"finding\": \"NuMA residues 1972–2007 constitute a nuclear localization signal (NLS); Lys1988 is essential for nuclear targeting; residues 1538–2115 are necessary and sufficient for spindle association; NuMA isoforms NuMA-m and NuMA-s (arising from alternative splicing) localize to centrosomes in interphase and spindle poles in mitosis rather than the nucleus.\",\n      \"method\": \"Linker scanning mutagenesis, chimeric protein expression, transfection in CHO cells, immunofluorescence\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — systematic domain mapping with point mutations and isoform analysis, single lab\",\n      \"pmids\": [\"7962183\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Galectin-3 associates with NuMA at spindle poles in an O-GlcNAcylation-dependent manner during metaphase; loss of Galectin-3 impairs spindle pole cohesion and stable NuMA localization at the spindle pole in epithelial cells.\",\n      \"method\": \"Co-immunoprecipitation, siRNA knockdown, immunofluorescence, experimental mouse model\",\n      \"journal\": \"Scientific reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — co-IP with loss-of-function phenotype in both cell lines and mouse model, single lab\",\n      \"pmids\": [\"28469279\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"In acentrosomal human cells, NuMA forms small microtubule asters at nuclear envelope breakdown that are assembled by dynein and NuMA's clustering activity; NuMA organizes radial MT arrays incorporating Eg5 to promote spindle bipolarization; in cells with centrosomes, NuMA also promotes the initial step of spindle bipolarization.\",\n      \"method\": \"siRNA depletion of centrosome components, live imaging, NuMA siRNA in acentrosomal cells, immunofluorescence\",\n      \"journal\": \"The EMBO journal\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — defined genetic manipulation with live imaging in acentrosomal system, single lab\",\n      \"pmids\": [\"31782546\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NuMA is a large coiled-coil nuclear protein that undergoes CDK1-mediated phosphorylation at mitotic entry to exit the nucleus and form a complex with cytoplasmic dynein and dynactin, which transports NuMA to spindle microtubule minus-ends where it acts as a minus-end cargo adaptor; at spindle poles NuMA self-assembles into an insoluble cross-linking matrix that focuses and anchors microtubule minus-ends, a process regulated by Aurora-A and Plk1 phosphorylation; at the cell cortex during metaphase, NuMA is recruited by the LGN/Gαi complex (itself activated by Ric-8A GEF activity) and directly by phosphoinositides, assembling dynein-dynactin clusters that generate pulling forces on astral microtubules to orient the spindle, with cortical NuMA levels precisely tuned by CDK1 (metaphase suppression), Aurora-A, Plk1, and PP2A activity; in anaphase, CDK1 inactivation allows LGN-independent, phosphoinositide-dependent cortical NuMA accumulation that drives spindle elongation; in interphase, NuMA resides in the nuclear matrix where it interacts with protein 4.1, p53, RNA Pol I, SNF2h, and 53BP1 to contribute to nuclear architecture, p53 target gene selectivity, rDNA transcription, chromatin remodeling at DNA breaks, and regulation of DNA repair pathway choice.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"NuMA is a large coiled-coil protein that functions as a master organizer of mitotic spindle architecture and a regulator of nuclear processes during interphase. During mitosis, CDK1 phosphorylation triggers NuMA's release from the nucleus; NuMA is then transported poleward by cytoplasmic dynein–dynactin, where it self-assembles into an insoluble cross-linking matrix that focuses microtubule minus-ends at spindle poles—a function essential for centrosome–spindle attachment under tension and for acentrosomal spindle bipolarization [PMID:8898198, PMID:19255246, PMID:31782546]. At the cell cortex, NuMA is recruited by LGN/Gαi and directly by phosphoinositides to cluster dynein–dynactin force generators that pull on astral microtubules for spindle orientation, with cortical NuMA levels dynamically tuned by CDK1, Aurora-A, and Plk1 phosphorylation and opposed by PP2A dephosphorylation [PMID:15537540, PMID:24996901, PMID:23921553, PMID:26832443]. In interphase, NuMA resides in the nuclear matrix where it contributes to p53 target-gene selectivity by recruiting CDK8/Mediator, facilitates rDNA transcription through RNA Pol I association, and regulates DNA double-strand break repair by controlling SNF2h-mediated chromatin remodeling and 53BP1 dynamics [PMID:23589328, PMID:28981686, PMID:24753406, PMID:30812030].\",\n  \"teleology\": [\n    {\n      \"year\": 1992,\n      \"claim\": \"Cloning NuMA established it as a large nuclear protein with a tripartite domain architecture—globular heads flanking a long coiled-coil rod—that dynamically redistributes between the nucleus and spindle poles during mitosis, framing the central question of how a nuclear protein reorganizes into a mitotic structural element.\",\n      \"evidence\": \"cDNA cloning, sequence analysis, and double immunofluorescence with anti-lamin antibodies in human cells\",\n      \"pmids\": [\"1541630\", \"1541636\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No functional data; structure-function relationship of domains unknown\", \"No interacting partners identified\"]\n    },\n    {\n      \"year\": 1995,\n      \"claim\": \"Immunodepletion and reconstitution experiments demonstrated that NuMA is essential for organizing microtubule asters and that its C-terminal tail mediates microtubule bundling, establishing NuMA as a direct structural organizer of mitotic spindle poles rather than a passive nuclear scaffold.\",\n      \"evidence\": \"Cell-free mitotic aster assembly, immunodepletion with recombinant protein rescue, EM of purified protein, CDK1 phosphosite mutagenesis\",\n      \"pmids\": [\"7593190\", \"7781599\", \"7769006\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Motor-dependent transport mechanism to poles not yet defined\", \"Regulation of NuMA–MT interaction beyond CDK1 unknown\"]\n    },\n    {\n      \"year\": 1999,\n      \"claim\": \"EM and self-assembly studies revealed that NuMA oligomerizes into multiarm structures through C-terminal interactions and forms an insoluble matrix at spindle poles independent of microtubules, explaining how NuMA provides a structural scaffold that anchors MT minus-ends.\",\n      \"evidence\": \"In vitro assembly with EM, biochemical fractionation, immunogold EM of spindle poles\",\n      \"pmids\": [\"10075938\", \"10098933\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Regulation of assembly/disassembly cycle unclear\", \"How the matrix integrates with pericentriolar material not resolved\"]\n    },\n    {\n      \"year\": 2000,\n      \"claim\": \"Live imaging and biochemistry established that dynein–dynactin actively transports NuMA to spindle poles after nuclear envelope breakdown, forming a mitosis-specific complex whose disruption blocks pole assembly—linking NuMA's motor-dependent transport to its structural role.\",\n      \"evidence\": \"Live GFP imaging, immunoprecipitation, gel filtration, dynamitin overexpression in mammalian cells\",\n      \"pmids\": [\"10811826\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Which NuMA domains contact dynein LIC not yet mapped\", \"Regulation of complex formation/dissociation unclear\"]\n    },\n    {\n      \"year\": 2004,\n      \"claim\": \"Discovery that LGN acts as a conformational switch binding both NuMA and Gαi resolved how NuMA reaches the cell cortex for spindle orientation, establishing the LGN/Gαi/NuMA ternary complex as the cortical force-generating pathway, with CDK1-mediated cyclin B degradation releasing NuMA from dynein at anaphase.\",\n      \"evidence\": \"FRET biosensor, yeast two-hybrid, RNAi, live imaging; Xenopus egg extract with non-degradable cyclin B\",\n      \"pmids\": [\"15537540\", \"14710193\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether LGN and direct MT binding are mutually exclusive in vivo not tested\", \"Cortical force magnitude not measured\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Conditional NuMA knockout in mice proved NuMA is essential in vivo for tethering spindle microtubules to centrosomes under tension, resolving the question of whether NuMA has a unique function beyond what centrosomes and other crosslinkers provide.\",\n      \"evidence\": \"Conditional loss-of-function allele in mouse primary cells, live imaging\",\n      \"pmids\": [\"19255246\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NuMA's tethering and cortical functions are independently essential in vivo not separated\", \"Tissue-specific requirements not fully explored\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Crystal structures of LGN bound to NuMA versus mInsc revealed mutually exclusive binding to the same TPR surface, establishing that Par3/mInsc/LGN and NuMA/LGN/Gαi act sequentially rather than simultaneously in orienting the spindle.\",\n      \"evidence\": \"X-ray crystallography with in vitro competition binding assays\",\n      \"pmids\": [\"21816348\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Temporal handoff mechanism from mInsc to NuMA in vivo not directly visualized\", \"Whether additional factors modulate the switch unknown\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Multiple groups established that CDK1 phosphorylation of NuMA T2055 suppresses cortical NuMA during metaphase while PP2A counteracts this, and that a C-terminal lipid-binding domain mediates LGN-independent cortical association in anaphase—revealing a phospho-switch coupling mitotic progression to spindle positioning forces.\",\n      \"evidence\": \"In vitro kinase assays, phosphomutant rescue, lipid-binding assays, live imaging in human cells and keratinocytes\",\n      \"pmids\": [\"23921553\", \"24371089\", \"24109598\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Precise PIP species specificity not determined\", \"How cortical NuMA domains are spatially restricted to the cell equator at anaphase unclear\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Direct binding of NuMA to phosphoinositides (PIP/PIP2) was demonstrated, and PIP depletion abolished cortical NuMA, establishing phosphoinositides as the primary cortical receptor for NuMA during anaphase when LGN/Gαi are dispensable.\",\n      \"evidence\": \"In vitro lipid-binding assay, chemical/enzymatic PIP depletion, immunofluorescence, siRNA rescue\",\n      \"pmids\": [\"24996901\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether PIP2 clustering or local PIP2 generation provides spatial specificity unknown\", \"Structural basis of NuMA–PIP2 interaction not resolved\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Aurora-A phosphorylation was shown to control NuMA's dynamic exchange between spindle poles and the cortex without affecting MT or LGN binding affinity, and in vivo mutagenesis of NuMA's MT-binding domain in mouse skin caused spindle orientation defects and neonatal lethality, demonstrating NuMA's direct MT engagement is essential for tissue morphogenesis.\",\n      \"evidence\": \"In vitro kinase assay, FRAP, phosphomutants; conditional MT-binding domain knockout in mouse epidermis\",\n      \"pmids\": [\"26832443\", \"26765568\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Aurora-A phosphorylation alters NuMA mobility mechanistically (conformational change?) not defined\", \"Relative contributions of dynein-mediated vs. direct MT-binding functions in different tissues not quantified\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Quantitative imaging and structural studies revealed that NuMA localizes to MT minus-ends independently of dynein and recruits dynactin there, while a crystal structure showed Importin-α/β masks NuMA's MT-binding region with RanGTP releasing the inhibition—establishing NuMA as a bona fide minus-end cargo adaptor regulated by the Ran gradient.\",\n      \"evidence\": \"Laser ablation, domain deletion rescue, X-ray crystallography of Importin-α/NuMA complex, in vitro MT-binding assay\",\n      \"pmids\": [\"29185983\", \"28939615\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether Ran gradient regulation operates at the cortex as well as at poles unclear\", \"Minus-end recognition mechanism (structural feature of minus-end) not identified\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Optogenetic cortical targeting showed that NuMA—not dynein alone—is sufficient to generate spindle-pulling forces by assembling cortical focal clusters of dynein–dynactin; Plk1 was identified as an additional kinase phosphorylating NuMA to tune cortical levels, adding a third kinase input to the cortical regulatory circuit.\",\n      \"evidence\": \"Light-induced cortical targeting with AID depletion and domain dissection; Plk1 in vitro kinase assay with pharmacological inhibition and FRAP\",\n      \"pmids\": [\"29848445\", \"30456393\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How CDK1, Aurora-A, and Plk1 phosphorylation events are integrated on the same NuMA molecule not resolved\", \"Stoichiometry of NuMA within cortical clusters unknown\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Structural determination of NuMA–LIC1/LIC2 interfaces identified two distinct dynein light-intermediate-chain-binding sites (hook domain and CC1-box), both essential for spindle positioning, providing the molecular basis for NuMA's function as a dynein cargo adaptor.\",\n      \"evidence\": \"X-ray crystallography of NuMA–LIC1/2 interfaces, mutagenesis, functional cell division assay\",\n      \"pmids\": [\"32413290\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the two LIC-binding sites engage one or two dynein complexes simultaneously unknown\", \"Regulation of LIC-binding site accessibility not addressed\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Discovery of NuMA's interphase nuclear functions revealed it binds p53 and recruits CDK8/Mediator selectively to p21 but not PUMA promoters, establishing NuMA as a transcriptional cofactor that confers target-gene selectivity to the p53 response.\",\n      \"evidence\": \"Co-immunoprecipitation, ChIP, siRNA knockdown with selective gene expression readout\",\n      \"pmids\": [\"23589328\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis for selective promoter targeting unknown\", \"Whether NuMA acts on other transcription factors besides p53 not tested\", \"Independent replication needed\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"NuMA was found to accumulate at DNA damage sites in a PARylation-dependent manner, interact with SNF2h to promote chromatin decompaction and homologous recombination, linking NuMA's nuclear matrix role to active DNA repair.\",\n      \"evidence\": \"Co-IP, FRAP, laser microirradiation, siRNA with chromosomal DSB repair assays\",\n      \"pmids\": [\"24753406\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether NuMA is directly PARylated at breaks or recruited via PAR binding unclear\", \"Relationship to NuMA's interaction with tankyrase-1 at spindle poles not addressed\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"NuMA was shown to sequester 53BP1 in undamaged nuclei and limit its access to breaks, with NuMA depletion altering PARP inhibitor sensitivity in BRCA1-null cells, revealing NuMA as a regulator of DNA repair pathway choice.\",\n      \"evidence\": \"Co-IP, FRAP, NHEJ assay, class-switch recombination assay, PARP inhibitor sensitivity\",\n      \"pmids\": [\"30812030\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Mechanism by which NuMA restrains 53BP1 mobility not structurally defined\", \"In vivo relevance for tumor suppression not tested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"NuMA was shown to undergo liquid–liquid phase separation at mitotic entry, regulated by Aurora-A, concentrating tubulins and the MT depolymerase Kif2A at poles to promote poleward flux—providing a biophysical framework for how NuMA's self-assembly creates a functional spindle pole compartment.\",\n      \"evidence\": \"In vitro LLPS reconstitution, live cell condensate imaging, phosphomimetic/phosphodead mutants, siRNA\",\n      \"pmids\": [\"34887424\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether LLPS and the previously described insoluble matrix represent the same or distinct states unclear\", \"Contribution of LLPS versus oligomeric cross-linking to pole integrity not separated\", \"Independent replication of LLPS findings needed\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Key unresolved questions include how the three mitotic kinase inputs (CDK1, Aurora-A, Plk1) are integrated on a single NuMA molecule to produce graded cortical versus polar localization, the structural basis of NuMA's phosphoinositide binding and minus-end recognition, and the extent to which NuMA's interphase nuclear functions (p53 cofactor, rDNA transcription, DNA repair pathway choice) are mechanistically independent or coupled through a common scaffolding principle.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Integrated phospho-regulation model lacking\", \"No structural data for NuMA–PIP2 or NuMA–minus-end interfaces\", \"Whether nuclear and mitotic NuMA pools are regulated by shared or distinct mechanisms unknown\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [0, 3, 7, 22, 29, 30]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [20, 32, 19]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [23, 31, 22]},\n      {\"term_id\": \"GO:0008289\", \"supporting_discovery_ids\": [15, 16]},\n      {\"term_id\": \"GO:0140110\", \"supporting_discovery_ids\": [37, 42]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [2, 46, 37, 42]},\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 1, 3, 19, 32]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 22, 23, 29]},\n      {\"term_id\": \"GO:0005886\", \"supporting_discovery_ids\": [6, 14, 15, 16, 23]},\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [42]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1, 3, 4, 14, 19, 22, 23]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [36, 38]},\n      {\"term_id\": \"R-HSA-74160\", \"supporting_discovery_ids\": [37, 42]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [6, 25, 26]}\n    ],\n    \"complexes\": [\n      \"NuMA–dynein–dynactin\",\n      \"LGN/Gαi/NuMA\",\n      \"NuMA–tankyrase-1\"\n    ],\n    \"partners\": [\n      \"DYNC1H1\",\n      \"DCTN1\",\n      \"GPSM2\",\n      \"GNAI1\",\n      \"AURKA\",\n      \"EPB41\",\n      \"TNKS\",\n      \"TP53\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}