{"gene":"NEDD1","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2005,"finding":"GCP-WD/NEDD1 is a subunit of the human γ-TuRC and acts as an attachment factor targeting the γ-TuRC to the centrosome and mitotic spindle; depletion of GCP-WD disperses the γ-TuRC from the centrosome and abolishes centrosomal microtubule nucleation, while leaving the γ-TuRC intact and able to bind microtubules. Mitotic phosphorylation of GCP-WD is separately required for γ-tubulin association with the spindle.","method":"siRNA depletion, dominant-negative inhibition, immunofluorescence, co-immunoprecipitation, microtubule nucleation assays in human cells","journal":"Nature Cell Biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP establishing γ-TuRC subunit identity, clean siRNA KD with defined centrosomal and spindle phenotypes, replicated by multiple subsequent studies","pmids":["16378099"],"is_preprint":false},{"year":2006,"finding":"NEDD1 is required for centrosomal targeting of the γ-TuRC and for centriole duplication; NEDD1 can target to the centrosome independently of γ-tubulin, but γ-tubulin cannot reach the centrosome without NEDD1. The C-terminal half of NEDD1 mediates binding to γ-TuRCs.","method":"siRNA depletion, GFP-fusion overexpression (dominant negative), immunofluorescence, centrosome microtubule nucleation assays in human cells","journal":"The Journal of Cell Biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — clean KD with multiple defined cellular phenotypes (nucleation, centriole duplication), orthogonal dominant-negative approach, independently confirmed","pmids":["16461362"],"is_preprint":false},{"year":2008,"finding":"FAM29A interacts with the NEDD1–γ-tubulin complex and recruits it to the mitotic spindle, promoting microtubule-dependent microtubule amplification and kinetochore fiber maturation.","method":"Co-immunoprecipitation, mass spectrometry, siRNA depletion, immunofluorescence, nocodazole washout microtubule regrowth assay","journal":"The Journal of Cell Biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP identifying complex, siRNA KD with defined spindle phenotype, orthogonal MS identification","pmids":["19029337"],"is_preprint":false},{"year":2008,"finding":"Xenopus NEDD1 exists in a complex distinct from the γ-TuRC and is largely dispensable for targeting γ-tubulin to centrosomes in Xenopus egg extracts, but is required for microtubule organization in those extracts.","method":"Immunodepletion of Xenopus egg extracts, sucrose gradient fractionation, microtubule nucleation/organization assays","journal":"Journal of Cell Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean immunodepletion with defined phenotype in Xenopus egg extracts, single lab, two orthogonal methods; note result partially contradicts human-cell data","pmids":["18252801"],"is_preprint":false},{"year":2009,"finding":"FAM29A is regulated by Plk1, and Plk1, FAM29A, and NEDD1 form three separate complexes in vivo. Plk1 recruits FAM29A to spindle microtubules, which in turn targets NEDD1 to the spindle; Plk1 independently recruits NEDD1 to centrosomes. FAM29A controls partitioning of NEDD1 between centrosomes and spindle.","method":"Co-immunoprecipitation, siRNA depletion, immunofluorescence, overexpression studies in mammalian cells","journal":"Journal of Cell Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP demonstrating separate complexes, depletion experiments with defined NEDD1 redistribution phenotype, single lab","pmids":["19596795"],"is_preprint":false},{"year":2010,"finding":"NEDD1 directly binds γ-tubulin through a 62-residue C-terminal helical domain that forms a stable tetramer in solution; mutation of residues in this domain disrupts γ-tubulin binding and causes mis-localization of γ-tubulin away from the centrosome.","method":"In vitro binding assays, site-directed mutagenesis, CD spectroscopy, analytical ultracentrifugation, immunofluorescence in mammalian cells","journal":"PLoS ONE","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro reconstitution of direct binding, mutagenesis validating binding site, structural characterization (helical tetramer), single lab","pmids":["20224777"],"is_preprint":false},{"year":2010,"finding":"NEDD1 is an essential component of acentriolar MTOCs in mouse oocytes, co-localizing with γ-tubulin and pericentrin in a pericentrin-dependent manner. NEDD1 knockdown reduces γ-tubulin at MTOCs, disrupts meiotic spindle structure, causes metaphase-I arrest via spindle checkpoint activation, and leads to high-frequency aneuploidy.","method":"siRNA knockdown in mouse oocytes, immunofluorescence, spindle checkpoint assay (MAD2 detection), chromosome segregation analysis","journal":"Developmental Biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean siRNA KD with defined meiotic phenotype, multiple readouts (spindle structure, γ-tubulin localization, SAC activation, aneuploidy), single lab","pmids":["20079731"],"is_preprint":false},{"year":2012,"finding":"Nek9 phosphorylates NEDD1 on Ser377, driving recruitment of NEDD1 and thereby γ-tubulin to the centrosome in mitotic cells. This role of Nek9 requires Plk1-dependent activation of Nek9 but is independent of downstream kinases Nek6 and Nek7.","method":"In vitro kinase assay, phospho-specific antibodies, mutagenesis, Xenopus egg extracts, siRNA depletion in mammalian cells, immunofluorescence","journal":"Current Biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay with mutagenesis, validated in Xenopus extracts and mammalian cells, epistasis analysis, single lab","pmids":["22818914"],"is_preprint":false},{"year":2012,"finding":"Multiple phosphorylation sites in the S557–S574 region of NEDD1, near its γ-tubulin-binding domain, finely tune the NEDD1–γ-tubulin interaction and spindle assembly. S565–S574 phosphorylation inhibits γ-tubulin binding; additional S557–T560 mutations restore binding. CEP192 associates with NEDD1 and modulates its mitotic phosphorylation.","method":"Mass spectrometry phosphosite mapping, serine-to-alanine mutagenesis, co-immunoprecipitation, siRNA rescue experiments, immunofluorescence","journal":"Journal of Cell Science","confidence":"High","confidence_rationale":"Tier 1 / Moderate — MS phosphosite identification plus mutagenesis rescue, co-IP of CEP192–NEDD1 complex, multiple orthogonal methods in single study","pmids":["22595525"],"is_preprint":false},{"year":2012,"finding":"Aurora A phosphorylates NEDD1 at Ser405, and this phosphorylation is specifically required for microtubule nucleation around chromosomes (not at centrosomes) and for RanGTP-driven aster formation in Xenopus egg extracts.","method":"In vitro kinase assay, phospho-specific antibodies, site-directed mutagenesis, Xenopus egg extract MT nucleation assays, siRNA rescue in human cells, immunofluorescence","journal":"Current Biology","confidence":"High","confidence_rationale":"Tier 1 / Strong — in vitro kinase assay with mutagenesis, validated in both Xenopus extracts and human cells, independently replicated (Courthéoux et al. 2019)","pmids":["23273898"],"is_preprint":false},{"year":2012,"finding":"Cep57 interacts with NEDD1, and this interaction is required for centrosomal localization of Cep57. Loss of Cep57 causes PCM fragmentation, multipolar spindles, and weakened centrosomal microtubule nucleation.","method":"Co-immunoprecipitation, siRNA depletion, immunofluorescence in mammalian cells","journal":"Cell Research","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — Co-IP demonstrating interaction, siRNA KD with defined phenotype, single lab, no reciprocal IP reported in abstract","pmids":["22508265"],"is_preprint":false},{"year":2019,"finding":"Aurora A phosphorylation of NEDD1 on Ser405 is required for NEDD1 concentration in the midzone during central spindle (cytokinetic spindle) assembly; a phosphomimetic NEDD1-S405E mutant rescues midzone microtubule nucleation under Aurora A inhibition.","method":"Aurora A inhibitor treatment, phosphomimetic/phospho-null NEDD1 mutant expression, immunofluorescence, microtubule nucleation assays in human cells","journal":"Journal of Cell Science","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — phosphomimetic rescue experiment, pharmacological inhibition, single lab, extends prior Ser405 finding to central spindle context","pmids":["31028180"],"is_preprint":false},{"year":2021,"finding":"PLK4 phosphorylates NEDD1 at Ser325, directly promoting NEDD1 binding to SAS-6 and recruitment of SAS-6 to the centrosome, thereby initiating cartwheel assembly and daughter centriole biogenesis. Phosphomimetic S325E promotes these initiations; non-phosphorylatable S325A abolishes them.","method":"In vitro kinase assay, co-immunoprecipitation, phosphomimetic/phospho-null mutagenesis, immunofluorescence, overexpression in human cells","journal":"The Journal of Cell Biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — in vitro kinase assay plus mutagenesis (gain- and loss-of-function), Co-IP of NEDD1–SAS-6 complex, multiple orthogonal methods, single lab","pmids":["33351100"],"is_preprint":false},{"year":2022,"finding":"NEDD1 phosphorylation at S411 is essential for MT branching nucleation on pre-existing microtubules, demonstrated directly by TIRF microscopy in Xenopus egg extracts. S411 phosphorylation also coordinates the balance between centrosome- and chromosome-dependent MT nucleation required for bipolar spindle assembly.","method":"Stable inducible HeLa cell lines expressing phospho-variants, Xenopus egg extract TIRF microscopy of MT branching, immunofluorescence","journal":"Biology Open","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct TIRF visualization of branching plus phospho-variant stable cell lines, single lab, single study","pmids":["36318115"],"is_preprint":false},{"year":2024,"finding":"Cryo-EM structures of NEDD1 bound to the human γ-TuRC reveal that the C-terminus of NEDD1 forms a tetrameric α-helical assembly contacting the lumen of the γ-TuRC cone, anchored to GCP4, GCP5, and GCP6 via MZT1–GCP3 subcomplexes, with its microtubule-binding WD40 domains oriented away from the complex. NEDD1 does not induce conformational changes in the γ-TuRC. CDK5RAP2 and NEDD1 can simultaneously associate with the open conformation of the γ-TuRC.","method":"Cryo-EM structure determination, AlphaFold modeling, biochemical pulldown of NEDD1 mutants from cultured cells","journal":"bioRxiv (preprint; published as JCB 2025 per PMID:39574704)","confidence":"High","confidence_rationale":"Tier 1 / Moderate — cryo-EM structure with biochemical mutagenesis validation, multiple orthogonal methods, preprint but rigorous structural study","pmids":["39574704"],"is_preprint":true},{"year":2025,"finding":"MZT1 inhibits NEDD1 ubiquitination, thereby stabilizing NEDD1 protein levels in gastric cancer cells; MZT1 depletion reduces NEDD1 protein, decreasing proliferation and sensitizing cells to glucose starvation.","method":"Ubiquitination assay, co-immunoprecipitation, siRNA knockdown, Western blot, in vivo xenograft","journal":"Life Sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — direct ubiquitination assay showing MZT1 suppresses NEDD1 ubiquitination, siRNA with defined phenotype, single lab","pmids":["40204068"],"is_preprint":false},{"year":2025,"finding":"The NEDD1 WD40 β-propeller domain binds directly inside the V-junction of Augmin, enhancing Augmin dimerization; this interaction, together with Augmin's dual CH-domain MT-binding, creates a platform for γ-TuRC recruitment and branched MT nucleation.","method":"Reconstitution of plant Augmin–NEDD1 complex, cryo-EM structure determination, crosslinking mass spectrometry, evolutionary covariation analysis","journal":"Nature Communications","confidence":"High","confidence_rationale":"Tier 1 / Moderate — reconstitution of complex, cryo-EM structure, crosslinking MS, evolutionary validation; note this uses plant Augmin but identifies conserved NEDD1 WD40 binding interface","pmids":["41387433"],"is_preprint":false},{"year":2023,"finding":"METTL3 promotes NEDD1 mRNA translation via YTHDF1-dependent m6A modification in DLBCL cells; NEDD1 in turn activates Hedgehog signaling to promote immune escape.","method":"meRIP-qPCR, dual-luciferase Hedgehog pathway assay, siRNA knockdown, overexpression, Western blot in DLBCL cell lines","journal":"Immunity, Inflammation and Disease","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway activation inferred from luciferase reporter without direct mechanistic link between NEDD1 and Hedgehog pathway components established","pmids":["36840486"],"is_preprint":false},{"year":2025,"finding":"In human airway multiciliated cells, NEDD1 localizes to the basal foot of ciliary basal bodies and is essential for basal foot-dependent microtubule organization; depletion of NEDD1 (but not ninein or HAUS) disrupts this microtubule organization.","method":"Expansion microscopy 3D mapping, siRNA depletion, immunofluorescence in human airway multiciliated cells","journal":"bioRxiv","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct localization by expansion microscopy with functional consequence from KD, single preprint, single lab","pmids":["bio_10.1101_2025.09.04.674302"],"is_preprint":true}],"current_model":"NEDD1/GCP-WD is a WD40-repeat adaptor protein that targets the γ-tubulin ring complex (γ-TuRC) to centrosomes, mitotic spindle microtubules, and basal bodies via a C-terminal tetrameric α-helical domain that contacts the γ-TuRC lumen (anchored through GCP4/5/6 via MZT1–GCP3 modules), while its N-terminal WD40 domain binds microtubules and Augmin; its recruitment and activity are orchestrated by site-specific phosphorylation events—Nek9 (S377) for centrosomal targeting, Aurora A (S405) for chromosomal and midzone MT nucleation, S411 for MT branching, and PLK4 (S325) for SAS-6 recruitment and centriole biogenesis—making NEDD1 a phosphorylation-regulated hub that coordinates all major γ-TuRC-dependent microtubule nucleation pathways during cell division."},"narrative":{"mechanistic_narrative":"NEDD1 (GCP-WD) is a WD40-repeat adaptor that targets the γ-tubulin ring complex (γ-TuRC) to microtubule-organizing sites and thereby controls γ-TuRC-dependent microtubule nucleation during cell division [PMID:16378099, PMID:16461362]. It is a γ-TuRC-associated subunit whose depletion disperses γ-tubulin from the centrosome and abolishes centrosomal microtubule nucleation while leaving the γ-TuRC otherwise intact; γ-tubulin cannot reach the centrosome without NEDD1, but NEDD1 localizes independently, establishing it as the upstream attachment factor and also as a requirement for centriole duplication [PMID:16378099, PMID:16461362]. Binding to the γ-TuRC is mediated by a C-terminal ~62-residue α-helical domain that directly contacts γ-tubulin and forms a stable tetramer [PMID:20224777]; cryo-EM shows this tetrameric helical assembly inserts into the lumen of the γ-TuRC cone, anchored to GCP4/5/6 through MZT1–GCP3 subcomplexes, while its N-terminal WD40 β-propeller domains project outward toward microtubules [PMID:39574704]. The WD40 domain binds directly inside the V-junction of Augmin, enhancing Augmin dimerization to create a platform for γ-TuRC recruitment and branched microtubule nucleation [PMID:41387433]. NEDD1 function is governed by site-specific phosphorylation that partitions it among distinct nucleation pathways: Nek9 phosphorylates Ser377 to drive centrosomal recruitment downstream of Plk1 [PMID:22818914], Aurora A phosphorylates Ser405 to enable chromosome-proximal and central-spindle/midzone nucleation [PMID:23273898, PMID:31028180], and PLK4 phosphorylates Ser325 to promote SAS-6 binding and cartwheel-dependent centriole biogenesis [PMID:33351100]; additional phosphosites near the γ-tubulin-binding domain tune the interaction and are modulated by CEP192 [PMID:22595525]. Beyond mitosis, NEDD1 is essential for acentriolar spindle organization in oocytes, where its loss causes meiotic spindle defects and aneuploidy [PMID:20079731], and it localizes to the basal foot of ciliary basal bodies to organize microtubules in multiciliated cells [PMID:bio_10.1101_2025.09.04.674302].","teleology":[{"year":2005,"claim":"Established that γ-TuRC recruitment to the centrosome requires a dedicated adaptor rather than being intrinsic to the complex, identifying NEDD1/GCP-WD as that attachment factor.","evidence":"siRNA depletion, dominant-negative inhibition, co-IP and microtubule nucleation assays in human cells","pmids":["16378099"],"confidence":"High","gaps":["Did not define the structural basis of γ-TuRC binding","Mitotic phosphorylation requirement noted but kinases unidentified"]},{"year":2006,"claim":"Resolved the directionality of the targeting relationship—NEDD1 localizes independently of γ-tubulin but γ-tubulin depends on NEDD1—and extended NEDD1 function to centriole duplication.","evidence":"siRNA depletion and GFP-fusion dominant-negative overexpression in human cells with nucleation/duplication readouts","pmids":["16461362"],"confidence":"High","gaps":["Mapped binding only to C-terminal half, not a defined motif","Mechanism linking NEDD1 to centriole duplication unresolved at this stage"]},{"year":2008,"claim":"Distinguished centrosomal from spindle-based recruitment by showing FAM29A (Augmin component) tethers the NEDD1–γ-tubulin complex to spindle microtubules for MT-dependent amplification.","evidence":"Co-IP, mass spectrometry, siRNA depletion and nocodazole washout regrowth assays","pmids":["19029337"],"confidence":"High","gaps":["Direct NEDD1–Augmin interface not yet defined","Did not address how partitioning is regulated"]},{"year":2008,"claim":"Probed conservation of the targeting role across systems, finding NEDD1 in a non-γ-TuRC complex in Xenopus extracts and dispensable for centrosomal γ-tubulin targeting there, indicating context-dependent function.","evidence":"Immunodepletion and sucrose gradient fractionation of Xenopus egg extracts with nucleation assays","pmids":["18252801"],"confidence":"Medium","gaps":["Partially contradicts human-cell targeting data","Basis of the species/system difference not resolved"]},{"year":2009,"claim":"Showed how NEDD1 distribution between centrosome and spindle is controlled, placing Plk1 upstream of both FAM29A-dependent spindle targeting and direct centrosomal recruitment.","evidence":"Co-IP defining three separate complexes plus siRNA depletion and immunofluorescence in mammalian cells","pmids":["19596795"],"confidence":"Medium","gaps":["Direct Plk1 phosphosites on NEDD1 not mapped here","Single lab"]},{"year":2010,"claim":"Defined the molecular basis of γ-tubulin binding by reconstituting a direct interaction through a 62-residue C-terminal helical domain that forms a tetramer.","evidence":"In vitro binding, mutagenesis, CD spectroscopy and analytical ultracentrifugation with cellular localization","pmids":["20224777"],"confidence":"High","gaps":["Quaternary arrangement within the intact γ-TuRC not visualized","Single lab"]},{"year":2010,"claim":"Extended NEDD1's role to acentriolar MTOCs, showing it is essential for meiotic spindle assembly and chromosome segregation fidelity in oocytes.","evidence":"siRNA knockdown in mouse oocytes with spindle, γ-tubulin, SAC and aneuploidy readouts","pmids":["20079731"],"confidence":"Medium","gaps":["Pericentrin-dependence shown by localization, not mechanism","Single lab"]},{"year":2012,"claim":"Identified the kinase and site (Nek9–Ser377) coupling Plk1 signaling to centrosomal NEDD1/γ-tubulin recruitment, separating it from the Nek6/Nek7 branch.","evidence":"In vitro kinase assay, phospho-specific antibodies, mutagenesis, Xenopus extracts and mammalian-cell depletion","pmids":["22818914"],"confidence":"High","gaps":["How Ser377 phosphorylation mechanistically enhances centrosome binding unclear","Single lab"]},{"year":2012,"claim":"Showed that a cluster of phosphosites near the γ-tubulin-binding domain fine-tunes the NEDD1–γ-tubulin interaction and that CEP192 modulates this mitotic phosphorylation.","evidence":"MS phosphosite mapping, serine-to-alanine mutagenesis, Co-IP and siRNA rescue in human cells","pmids":["22595525"],"confidence":"High","gaps":["Responsible kinase(s) for the S557–S574 cluster not assigned","Single lab"]},{"year":2012,"claim":"Defined a spatially restricted nucleation pathway by showing Aurora A–Ser405 phosphorylation is specifically required for chromosome-proximal, RanGTP-driven nucleation but not centrosomal nucleation.","evidence":"In vitro kinase assay, phospho-specific antibodies, mutagenesis, Xenopus extract and human-cell rescue","pmids":["23273898"],"confidence":"High","gaps":["How Ser405 selectively licenses non-centrosomal nucleation unresolved"]},{"year":2012,"claim":"Linked NEDD1 to PCM integrity by showing it recruits Cep57 to centrosomes, whose loss fragments the PCM and weakens nucleation.","evidence":"Co-IP and siRNA depletion with immunofluorescence in mammalian cells","pmids":["22508265"],"confidence":"Medium","gaps":["No reciprocal IP reported","Direct binding interface undefined","Single lab"]},{"year":2019,"claim":"Extended the Aurora A–Ser405 axis to cytokinesis, showing this phosphorylation concentrates NEDD1 at the midzone for central spindle microtubule nucleation.","evidence":"Aurora A inhibition with phosphomimetic/phospho-null NEDD1 rescue in human cells","pmids":["31028180"],"confidence":"Medium","gaps":["Midzone recruitment partner not identified","Single lab"]},{"year":2021,"claim":"Connected NEDD1 phosphorylation to centriole biogenesis by showing PLK4–Ser325 phosphorylation drives SAS-6 binding and cartwheel initiation.","evidence":"In vitro kinase assay, Co-IP, phosphomimetic/phospho-null mutagenesis and overexpression in human cells","pmids":["33351100"],"confidence":"High","gaps":["Structural basis of phospho-NEDD1–SAS-6 binding not determined","Single lab"]},{"year":2022,"claim":"Directly visualized that Ser411 phosphorylation is required for branched microtubule nucleation and balances centrosome- versus chromosome-driven nucleation in spindle assembly.","evidence":"Phospho-variant stable HeLa lines and TIRF microscopy of MT branching in Xenopus egg extracts","pmids":["36318115"],"confidence":"Medium","gaps":["Responsible kinase for Ser411 not identified","Single lab"]},{"year":2024,"claim":"Provided the structural model for how NEDD1 docks onto the γ-TuRC, showing a C-terminal tetrameric helical bundle in the cone lumen anchored via MZT1–GCP3 to GCP4/5/6 with WD40 domains projecting outward.","evidence":"Cryo-EM, AlphaFold modeling and biochemical pulldowns of NEDD1 mutants (preprint, published JCB 2025)","pmids":["39574704"],"confidence":"High","gaps":["How phosphorylation modulates this docked state structurally unaddressed","Co-occupancy with CDK5RAP2 functional consequence unclear"]},{"year":2025,"claim":"Defined the WD40-domain interaction with Augmin at atomic resolution, showing it binds inside the Augmin V-junction and enhances dimerization to build a branching nucleation platform.","evidence":"Reconstitution of plant Augmin–NEDD1 complex, cryo-EM, crosslinking MS and evolutionary covariation","pmids":["41387433"],"confidence":"High","gaps":["Demonstrated with plant Augmin; full human complex not reconstituted","Interplay with branching-specific phosphosites not tested"]},{"year":2025,"claim":"Identified a post-translational stability control whereby MZT1 suppresses NEDD1 ubiquitination, linking NEDD1 abundance to cancer cell proliferation and metabolic stress sensitivity.","evidence":"Ubiquitination assay, Co-IP, siRNA knockdown, Western blot and xenograft in gastric cancer cells","pmids":["40204068"],"confidence":"Medium","gaps":["E3 ligase mediating NEDD1 ubiquitination not identified","Single lab"]},{"year":2025,"claim":"Extended NEDD1's microtubule-organizing role to differentiated cilia, localizing it to the basal foot of basal bodies where it is uniquely required for basal foot microtubule organization.","evidence":"Expansion microscopy 3D mapping and siRNA depletion in human airway multiciliated cells (preprint)","pmids":["bio_10.1101_2025.09.04.674302"],"confidence":"Medium","gaps":["Preprint; not peer-reviewed","Recruitment mechanism to basal foot undefined"]},{"year":null,"claim":"How the distinct phosphorylation events (Ser325, Ser377, Ser405, Ser411 and the S557–S574 cluster) are spatially and temporally integrated on a single NEDD1 molecule to switch between centrosomal, chromosomal, midzone, branching and centriolar nucleation pathways remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No single study resolves combinatorial phospho-state of NEDD1 in vivo","Kinases for several sites (S411, S557–S574 cluster) not all assigned","Structural effect of phosphorylation on γ-TuRC/Augmin docking not determined"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,1,5,14,16]},{"term_id":"GO:0008092","term_label":"cytoskeletal protein binding","supporting_discovery_ids":[5,14,16]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,1,7,10]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[0,2,13]},{"term_id":"GO:0005929","term_label":"cilium","supporting_discovery_ids":[18]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1,6,9,12]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[1,12,18]}],"complexes":["γ-tubulin ring complex (γ-TuRC)","Augmin/HAUS complex"],"partners":["TUBG1","FAM29A/HAUS6","MZT1","CEP192","CEP57","SAS-6","GCP4/TUBGCP4","GCP6/TUBGCP6"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8NHV4","full_name":"Protein NEDD1","aliases":["Neural precursor cell expressed developmentally down-regulated protein 1","NEDD-1"],"length_aa":660,"mass_kda":72.0,"function":"Required for mitosis progression. Promotes the nucleation of microtubules from the spindle","subcellular_location":"Cytoplasm, cytoskeleton, microtubule organizing center, centrosome","url":"https://www.uniprot.org/uniprotkb/Q8NHV4/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/NEDD1","classification":"Common Essential","n_dependent_lines":1204,"n_total_lines":1208,"dependency_fraction":0.9966887417218543},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000139350","cell_line_id":"CID000055","localizations":[{"compartment":"centrosome","grade":3},{"compartment":"cytoplasmic","grade":2}],"interactors":[{"gene":"MZT2B;MZT2A","stoichiometry":4.0},{"gene":"SACM1L","stoichiometry":0.2},{"gene":"TUBG1","stoichiometry":0.2},{"gene":"UBA1","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/target/CID000055","total_profiled":1310},"omim":[{"mim_id":"613433","title":"HAUS AUGMIN-LIKE COMPLEX, SUBUNIT 6; HAUS6","url":"https://www.omim.org/entry/613433"},{"mim_id":"600372","title":"NEURAL PRECURSOR CELL EXPRESSED, DEVELOPMENTALLY DOWNREGULATED 1; NEDD1","url":"https://www.omim.org/entry/600372"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Centrosome","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NEDD1"},"hgnc":{"alias_symbol":["GCP-WD","TUBGCP7"],"prev_symbol":[]},"alphafold":{"accession":"Q8NHV4","domains":[{"cath_id":"2.130.10.10","chopping":"5-304","consensus_level":"medium","plddt":92.4154,"start":5,"end":304},{"cath_id":"1.20.5","chopping":"567-660","consensus_level":"medium","plddt":80.4739,"start":567,"end":660}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8NHV4","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8NHV4-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8NHV4-F1-predicted_aligned_error_v6.png","plddt_mean":65.88},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NEDD1","jax_strain_url":"https://www.jax.org/strain/search?query=NEDD1"},"sequence":{"accession":"Q8NHV4","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8NHV4.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8NHV4/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8NHV4"}},"corpus_meta":[{"pmid":"16378099","id":"PMC_16378099","title":"GCP-WD is a gamma-tubulin targeting factor required for centrosomal and chromatin-mediated microtubule nucleation.","date":"2005","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16378099","citation_count":271,"is_preprint":false},{"pmid":"16461362","id":"PMC_16461362","title":"NEDD1-dependent recruitment of the gamma-tubulin ring complex to the centrosome is necessary for centriole duplication and spindle assembly.","date":"2006","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16461362","citation_count":230,"is_preprint":false},{"pmid":"19029337","id":"PMC_19029337","title":"FAM29A promotes microtubule amplification via recruitment of the NEDD1-gamma-tubulin complex to the mitotic spindle.","date":"2008","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/19029337","citation_count":95,"is_preprint":false},{"pmid":"22818914","id":"PMC_22818914","title":"Nek9 phosphorylation of NEDD1/GCP-WD contributes to Plk1 control of γ-tubulin recruitment to the mitotic centrosome.","date":"2012","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/22818914","citation_count":68,"is_preprint":false},{"pmid":"23273898","id":"PMC_23273898","title":"The role of NEDD1 phosphorylation by Aurora A in chromosomal microtubule nucleation and spindle function.","date":"2012","source":"Current biology : CB","url":"https://pubmed.ncbi.nlm.nih.gov/23273898","citation_count":54,"is_preprint":false},{"pmid":"20079731","id":"PMC_20079731","title":"NEDD1 is crucial for meiotic spindle stability and accurate chromosome segregation in mammalian oocytes.","date":"2010","source":"Developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/20079731","citation_count":52,"is_preprint":false},{"pmid":"20224777","id":"PMC_20224777","title":"A direct interaction with NEDD1 regulates gamma-tubulin recruitment to the centrosome.","date":"2010","source":"PloS 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microtubule nucleation, spindle assembly and beyond.","date":"2006","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/17005434","citation_count":29,"is_preprint":false},{"pmid":"18252801","id":"PMC_18252801","title":"Xenopus NEDD1 is required for microtubule organization in Xenopus egg extracts.","date":"2008","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/18252801","citation_count":26,"is_preprint":false},{"pmid":"26633906","id":"PMC_26633906","title":"Loss of γ-tubulin, GCP-WD/NEDD1 and CDK5RAP2 from the Centrosome of Neurons in Developing Mouse Cerebral and Cerebellar Cortex.","date":"2015","source":"Acta histochemica et cytochemica","url":"https://pubmed.ncbi.nlm.nih.gov/26633906","citation_count":23,"is_preprint":false},{"pmid":"19596795","id":"PMC_19596795","title":"FAM29A, a target of Plk1 regulation, controls the partitioning of NEDD1 between the mitotic spindle and the centrosomes.","date":"2009","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/19596795","citation_count":21,"is_preprint":false},{"pmid":"19243593","id":"PMC_19243593","title":"The centrosome protein NEDD1 as a potential pharmacological target to induce cell cycle arrest.","date":"2009","source":"Molecular cancer","url":"https://pubmed.ncbi.nlm.nih.gov/19243593","citation_count":20,"is_preprint":false},{"pmid":"23106787","id":"PMC_23106787","title":"Intraperitoneal delivery of a small interfering RNA targeting NEDD1 prolongs the survival of scirrhous gastric cancer model mice.","date":"2012","source":"Cancer science","url":"https://pubmed.ncbi.nlm.nih.gov/23106787","citation_count":19,"is_preprint":false},{"pmid":"33351100","id":"PMC_33351100","title":"PLK4-phosphorylated NEDD1 facilitates cartwheel assembly and centriole biogenesis initiations.","date":"2021","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/33351100","citation_count":16,"is_preprint":false},{"pmid":"18239929","id":"PMC_18239929","title":"Nedd1 expression as a marker of dynamic centrosomal localization during mouse embryonic development.","date":"2008","source":"Histochemistry and cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/18239929","citation_count":14,"is_preprint":false},{"pmid":"31028180","id":"PMC_31028180","title":"Microtubule nucleation during central spindle assembly requires NEDD1 phosphorylation on serine 405 by Aurora A.","date":"2019","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/31028180","citation_count":14,"is_preprint":false},{"pmid":"20150915","id":"PMC_20150915","title":"An essential function for the centrosomal protein NEDD1 in zebrafish development.","date":"2010","source":"Cell death and differentiation","url":"https://pubmed.ncbi.nlm.nih.gov/20150915","citation_count":14,"is_preprint":false},{"pmid":"7814034","id":"PMC_7814034","title":"Assignment of the developmentally regulated gene NEDD1 to human chromosome 12q22 by fluorescence in situ hybridization.","date":"1995","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/7814034","citation_count":6,"is_preprint":false},{"pmid":"38577589","id":"PMC_38577589","title":"NEDD1 overexpression increases cell proliferation, tumor immune escape, and drug resistance in LUAD.","date":"2024","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/38577589","citation_count":5,"is_preprint":false},{"pmid":"36840486","id":"PMC_36840486","title":"METTL3 enhances the effect of YTHDF1 on NEDD1 mRNA stability by m6A modification in diffuse large B-cell lymphoma cells.","date":"2023","source":"Immunity, inflammation and disease","url":"https://pubmed.ncbi.nlm.nih.gov/36840486","citation_count":5,"is_preprint":false},{"pmid":"36318115","id":"PMC_36318115","title":"NEDD1-S411 phosphorylation plays a critical function in the coordination of microtubule nucleation during mitosis.","date":"2022","source":"Biology open","url":"https://pubmed.ncbi.nlm.nih.gov/36318115","citation_count":4,"is_preprint":false},{"pmid":"39574704","id":"PMC_39574704","title":"Structure of the microtubule anchoring factor NEDD1 bound to the γ-tubulin ring complex.","date":"2024","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/39574704","citation_count":1,"is_preprint":false},{"pmid":"40204068","id":"PMC_40204068","title":"MZT1 protects gastric cancer against glucose starvation through targeting NEDD1.","date":"2025","source":"Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/40204068","citation_count":1,"is_preprint":false},{"pmid":"40034650","id":"PMC_40034650","title":"Cryo-EM structures of the Plant Augmin reveal its intertwined coiled-coil assembly, antiparallel dimerization and NEDD1 binding mechanisms.","date":"2025","source":"bioRxiv : the preprint server for biology","url":"https://pubmed.ncbi.nlm.nih.gov/40034650","citation_count":1,"is_preprint":false},{"pmid":"41387433","id":"PMC_41387433","title":"Cryo-EM structures of plant Augmin reveal coiled-coil assembly, antiparallel dimerization, and NEDD1 binding.","date":"2025","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/41387433","citation_count":0,"is_preprint":false},{"pmid":"40663060","id":"PMC_40663060","title":"Microtubule nucleation: How the NEDD1:MZT1:GCP3 trio captures the γ-TuRC.","date":"2025","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/40663060","citation_count":0,"is_preprint":false},{"pmid":null,"id":"bio_10.1101_2025.09.04.674302","title":"Microtubule organization and molecular architecture of ciliary basal bodies in multiciliated airway cells","date":"2025-09-04","source":"bioRxiv","url":"https://doi.org/10.1101/2025.09.04.674302","citation_count":0,"is_preprint":true}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14959,"output_tokens":4839,"usd":0.058731,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":13053,"output_tokens":5371,"usd":0.09977,"stage2_stop_reason":"end_turn"},"total_usd":0.158501,"stage1_batch_id":"msgbatch_013a8chtv3AWyTqv4z4sEF4C","stage2_batch_id":"msgbatch_01AHsuW63vinsvuTgzmQ3n62","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2005,\n      \"finding\": \"GCP-WD/NEDD1 is a subunit of the human γ-TuRC and acts as an attachment factor targeting the γ-TuRC to the centrosome and mitotic spindle; depletion of GCP-WD disperses the γ-TuRC from the centrosome and abolishes centrosomal microtubule nucleation, while leaving the γ-TuRC intact and able to bind microtubules. Mitotic phosphorylation of GCP-WD is separately required for γ-tubulin association with the spindle.\",\n      \"method\": \"siRNA depletion, dominant-negative inhibition, immunofluorescence, co-immunoprecipitation, microtubule nucleation assays in human cells\",\n      \"journal\": \"Nature Cell Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP establishing γ-TuRC subunit identity, clean siRNA KD with defined centrosomal and spindle phenotypes, replicated by multiple subsequent studies\",\n      \"pmids\": [\"16378099\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2006,\n      \"finding\": \"NEDD1 is required for centrosomal targeting of the γ-TuRC and for centriole duplication; NEDD1 can target to the centrosome independently of γ-tubulin, but γ-tubulin cannot reach the centrosome without NEDD1. The C-terminal half of NEDD1 mediates binding to γ-TuRCs.\",\n      \"method\": \"siRNA depletion, GFP-fusion overexpression (dominant negative), immunofluorescence, centrosome microtubule nucleation assays in human cells\",\n      \"journal\": \"The Journal of Cell Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — clean KD with multiple defined cellular phenotypes (nucleation, centriole duplication), orthogonal dominant-negative approach, independently confirmed\",\n      \"pmids\": [\"16461362\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"FAM29A interacts with the NEDD1–γ-tubulin complex and recruits it to the mitotic spindle, promoting microtubule-dependent microtubule amplification and kinetochore fiber maturation.\",\n      \"method\": \"Co-immunoprecipitation, mass spectrometry, siRNA depletion, immunofluorescence, nocodazole washout microtubule regrowth assay\",\n      \"journal\": \"The Journal of Cell Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP identifying complex, siRNA KD with defined spindle phenotype, orthogonal MS identification\",\n      \"pmids\": [\"19029337\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2008,\n      \"finding\": \"Xenopus NEDD1 exists in a complex distinct from the γ-TuRC and is largely dispensable for targeting γ-tubulin to centrosomes in Xenopus egg extracts, but is required for microtubule organization in those extracts.\",\n      \"method\": \"Immunodepletion of Xenopus egg extracts, sucrose gradient fractionation, microtubule nucleation/organization assays\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean immunodepletion with defined phenotype in Xenopus egg extracts, single lab, two orthogonal methods; note result partially contradicts human-cell data\",\n      \"pmids\": [\"18252801\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"FAM29A is regulated by Plk1, and Plk1, FAM29A, and NEDD1 form three separate complexes in vivo. Plk1 recruits FAM29A to spindle microtubules, which in turn targets NEDD1 to the spindle; Plk1 independently recruits NEDD1 to centrosomes. FAM29A controls partitioning of NEDD1 between centrosomes and spindle.\",\n      \"method\": \"Co-immunoprecipitation, siRNA depletion, immunofluorescence, overexpression studies in mammalian cells\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP demonstrating separate complexes, depletion experiments with defined NEDD1 redistribution phenotype, single lab\",\n      \"pmids\": [\"19596795\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"NEDD1 directly binds γ-tubulin through a 62-residue C-terminal helical domain that forms a stable tetramer in solution; mutation of residues in this domain disrupts γ-tubulin binding and causes mis-localization of γ-tubulin away from the centrosome.\",\n      \"method\": \"In vitro binding assays, site-directed mutagenesis, CD spectroscopy, analytical ultracentrifugation, immunofluorescence in mammalian cells\",\n      \"journal\": \"PLoS ONE\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro reconstitution of direct binding, mutagenesis validating binding site, structural characterization (helical tetramer), single lab\",\n      \"pmids\": [\"20224777\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"NEDD1 is an essential component of acentriolar MTOCs in mouse oocytes, co-localizing with γ-tubulin and pericentrin in a pericentrin-dependent manner. NEDD1 knockdown reduces γ-tubulin at MTOCs, disrupts meiotic spindle structure, causes metaphase-I arrest via spindle checkpoint activation, and leads to high-frequency aneuploidy.\",\n      \"method\": \"siRNA knockdown in mouse oocytes, immunofluorescence, spindle checkpoint assay (MAD2 detection), chromosome segregation analysis\",\n      \"journal\": \"Developmental Biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean siRNA KD with defined meiotic phenotype, multiple readouts (spindle structure, γ-tubulin localization, SAC activation, aneuploidy), single lab\",\n      \"pmids\": [\"20079731\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Nek9 phosphorylates NEDD1 on Ser377, driving recruitment of NEDD1 and thereby γ-tubulin to the centrosome in mitotic cells. This role of Nek9 requires Plk1-dependent activation of Nek9 but is independent of downstream kinases Nek6 and Nek7.\",\n      \"method\": \"In vitro kinase assay, phospho-specific antibodies, mutagenesis, Xenopus egg extracts, siRNA depletion in mammalian cells, immunofluorescence\",\n      \"journal\": \"Current Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay with mutagenesis, validated in Xenopus extracts and mammalian cells, epistasis analysis, single lab\",\n      \"pmids\": [\"22818914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Multiple phosphorylation sites in the S557–S574 region of NEDD1, near its γ-tubulin-binding domain, finely tune the NEDD1–γ-tubulin interaction and spindle assembly. S565–S574 phosphorylation inhibits γ-tubulin binding; additional S557–T560 mutations restore binding. CEP192 associates with NEDD1 and modulates its mitotic phosphorylation.\",\n      \"method\": \"Mass spectrometry phosphosite mapping, serine-to-alanine mutagenesis, co-immunoprecipitation, siRNA rescue experiments, immunofluorescence\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — MS phosphosite identification plus mutagenesis rescue, co-IP of CEP192–NEDD1 complex, multiple orthogonal methods in single study\",\n      \"pmids\": [\"22595525\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Aurora A phosphorylates NEDD1 at Ser405, and this phosphorylation is specifically required for microtubule nucleation around chromosomes (not at centrosomes) and for RanGTP-driven aster formation in Xenopus egg extracts.\",\n      \"method\": \"In vitro kinase assay, phospho-specific antibodies, site-directed mutagenesis, Xenopus egg extract MT nucleation assays, siRNA rescue in human cells, immunofluorescence\",\n      \"journal\": \"Current Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Strong — in vitro kinase assay with mutagenesis, validated in both Xenopus extracts and human cells, independently replicated (Courthéoux et al. 2019)\",\n      \"pmids\": [\"23273898\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"Cep57 interacts with NEDD1, and this interaction is required for centrosomal localization of Cep57. Loss of Cep57 causes PCM fragmentation, multipolar spindles, and weakened centrosomal microtubule nucleation.\",\n      \"method\": \"Co-immunoprecipitation, siRNA depletion, immunofluorescence in mammalian cells\",\n      \"journal\": \"Cell Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — Co-IP demonstrating interaction, siRNA KD with defined phenotype, single lab, no reciprocal IP reported in abstract\",\n      \"pmids\": [\"22508265\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"Aurora A phosphorylation of NEDD1 on Ser405 is required for NEDD1 concentration in the midzone during central spindle (cytokinetic spindle) assembly; a phosphomimetic NEDD1-S405E mutant rescues midzone microtubule nucleation under Aurora A inhibition.\",\n      \"method\": \"Aurora A inhibitor treatment, phosphomimetic/phospho-null NEDD1 mutant expression, immunofluorescence, microtubule nucleation assays in human cells\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — phosphomimetic rescue experiment, pharmacological inhibition, single lab, extends prior Ser405 finding to central spindle context\",\n      \"pmids\": [\"31028180\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"PLK4 phosphorylates NEDD1 at Ser325, directly promoting NEDD1 binding to SAS-6 and recruitment of SAS-6 to the centrosome, thereby initiating cartwheel assembly and daughter centriole biogenesis. Phosphomimetic S325E promotes these initiations; non-phosphorylatable S325A abolishes them.\",\n      \"method\": \"In vitro kinase assay, co-immunoprecipitation, phosphomimetic/phospho-null mutagenesis, immunofluorescence, overexpression in human cells\",\n      \"journal\": \"The Journal of Cell Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — in vitro kinase assay plus mutagenesis (gain- and loss-of-function), Co-IP of NEDD1–SAS-6 complex, multiple orthogonal methods, single lab\",\n      \"pmids\": [\"33351100\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"NEDD1 phosphorylation at S411 is essential for MT branching nucleation on pre-existing microtubules, demonstrated directly by TIRF microscopy in Xenopus egg extracts. S411 phosphorylation also coordinates the balance between centrosome- and chromosome-dependent MT nucleation required for bipolar spindle assembly.\",\n      \"method\": \"Stable inducible HeLa cell lines expressing phospho-variants, Xenopus egg extract TIRF microscopy of MT branching, immunofluorescence\",\n      \"journal\": \"Biology Open\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct TIRF visualization of branching plus phospho-variant stable cell lines, single lab, single study\",\n      \"pmids\": [\"36318115\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"Cryo-EM structures of NEDD1 bound to the human γ-TuRC reveal that the C-terminus of NEDD1 forms a tetrameric α-helical assembly contacting the lumen of the γ-TuRC cone, anchored to GCP4, GCP5, and GCP6 via MZT1–GCP3 subcomplexes, with its microtubule-binding WD40 domains oriented away from the complex. NEDD1 does not induce conformational changes in the γ-TuRC. CDK5RAP2 and NEDD1 can simultaneously associate with the open conformation of the γ-TuRC.\",\n      \"method\": \"Cryo-EM structure determination, AlphaFold modeling, biochemical pulldown of NEDD1 mutants from cultured cells\",\n      \"journal\": \"bioRxiv (preprint; published as JCB 2025 per PMID:39574704)\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — cryo-EM structure with biochemical mutagenesis validation, multiple orthogonal methods, preprint but rigorous structural study\",\n      \"pmids\": [\"39574704\"],\n      \"is_preprint\": true\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"MZT1 inhibits NEDD1 ubiquitination, thereby stabilizing NEDD1 protein levels in gastric cancer cells; MZT1 depletion reduces NEDD1 protein, decreasing proliferation and sensitizing cells to glucose starvation.\",\n      \"method\": \"Ubiquitination assay, co-immunoprecipitation, siRNA knockdown, Western blot, in vivo xenograft\",\n      \"journal\": \"Life Sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — direct ubiquitination assay showing MZT1 suppresses NEDD1 ubiquitination, siRNA with defined phenotype, single lab\",\n      \"pmids\": [\"40204068\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"The NEDD1 WD40 β-propeller domain binds directly inside the V-junction of Augmin, enhancing Augmin dimerization; this interaction, together with Augmin's dual CH-domain MT-binding, creates a platform for γ-TuRC recruitment and branched MT nucleation.\",\n      \"method\": \"Reconstitution of plant Augmin–NEDD1 complex, cryo-EM structure determination, crosslinking mass spectrometry, evolutionary covariation analysis\",\n      \"journal\": \"Nature Communications\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — reconstitution of complex, cryo-EM structure, crosslinking MS, evolutionary validation; note this uses plant Augmin but identifies conserved NEDD1 WD40 binding interface\",\n      \"pmids\": [\"41387433\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"METTL3 promotes NEDD1 mRNA translation via YTHDF1-dependent m6A modification in DLBCL cells; NEDD1 in turn activates Hedgehog signaling to promote immune escape.\",\n      \"method\": \"meRIP-qPCR, dual-luciferase Hedgehog pathway assay, siRNA knockdown, overexpression, Western blot in DLBCL cell lines\",\n      \"journal\": \"Immunity, Inflammation and Disease\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway activation inferred from luciferase reporter without direct mechanistic link between NEDD1 and Hedgehog pathway components established\",\n      \"pmids\": [\"36840486\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"In human airway multiciliated cells, NEDD1 localizes to the basal foot of ciliary basal bodies and is essential for basal foot-dependent microtubule organization; depletion of NEDD1 (but not ninein or HAUS) disrupts this microtubule organization.\",\n      \"method\": \"Expansion microscopy 3D mapping, siRNA depletion, immunofluorescence in human airway multiciliated cells\",\n      \"journal\": \"bioRxiv\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct localization by expansion microscopy with functional consequence from KD, single preprint, single lab\",\n      \"pmids\": [\"bio_10.1101_2025.09.04.674302\"],\n      \"is_preprint\": true\n    }\n  ],\n  \"current_model\": \"NEDD1/GCP-WD is a WD40-repeat adaptor protein that targets the γ-tubulin ring complex (γ-TuRC) to centrosomes, mitotic spindle microtubules, and basal bodies via a C-terminal tetrameric α-helical domain that contacts the γ-TuRC lumen (anchored through GCP4/5/6 via MZT1–GCP3 modules), while its N-terminal WD40 domain binds microtubules and Augmin; its recruitment and activity are orchestrated by site-specific phosphorylation events—Nek9 (S377) for centrosomal targeting, Aurora A (S405) for chromosomal and midzone MT nucleation, S411 for MT branching, and PLK4 (S325) for SAS-6 recruitment and centriole biogenesis—making NEDD1 a phosphorylation-regulated hub that coordinates all major γ-TuRC-dependent microtubule nucleation pathways during cell division.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NEDD1 (GCP-WD) is a WD40-repeat adaptor that targets the γ-tubulin ring complex (γ-TuRC) to microtubule-organizing sites and thereby controls γ-TuRC-dependent microtubule nucleation during cell division [#0, #1]. It is a γ-TuRC-associated subunit whose depletion disperses γ-tubulin from the centrosome and abolishes centrosomal microtubule nucleation while leaving the γ-TuRC otherwise intact; γ-tubulin cannot reach the centrosome without NEDD1, but NEDD1 localizes independently, establishing it as the upstream attachment factor and also as a requirement for centriole duplication [#0, #1]. Binding to the γ-TuRC is mediated by a C-terminal ~62-residue α-helical domain that directly contacts γ-tubulin and forms a stable tetramer [#5]; cryo-EM shows this tetrameric helical assembly inserts into the lumen of the γ-TuRC cone, anchored to GCP4/5/6 through MZT1–GCP3 subcomplexes, while its N-terminal WD40 β-propeller domains project outward toward microtubules [#14]. The WD40 domain binds directly inside the V-junction of Augmin, enhancing Augmin dimerization to create a platform for γ-TuRC recruitment and branched microtubule nucleation [#16]. NEDD1 function is governed by site-specific phosphorylation that partitions it among distinct nucleation pathways: Nek9 phosphorylates Ser377 to drive centrosomal recruitment downstream of Plk1 [#7], Aurora A phosphorylates Ser405 to enable chromosome-proximal and central-spindle/midzone nucleation [#9, #11], and PLK4 phosphorylates Ser325 to promote SAS-6 binding and cartwheel-dependent centriole biogenesis [#12]; additional phosphosites near the γ-tubulin-binding domain tune the interaction and are modulated by CEP192 [#8]. Beyond mitosis, NEDD1 is essential for acentriolar spindle organization in oocytes, where its loss causes meiotic spindle defects and aneuploidy [#6], and it localizes to the basal foot of ciliary basal bodies to organize microtubules in multiciliated cells [#18].\",\n  \"teleology\": [\n    {\n      \"year\": 2005,\n      \"claim\": \"Established that γ-TuRC recruitment to the centrosome requires a dedicated adaptor rather than being intrinsic to the complex, identifying NEDD1/GCP-WD as that attachment factor.\",\n      \"evidence\": \"siRNA depletion, dominant-negative inhibition, co-IP and microtubule nucleation assays in human cells\",\n      \"pmids\": [\"16378099\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not define the structural basis of γ-TuRC binding\", \"Mitotic phosphorylation requirement noted but kinases unidentified\"]\n    },\n    {\n      \"year\": 2006,\n      \"claim\": \"Resolved the directionality of the targeting relationship—NEDD1 localizes independently of γ-tubulin but γ-tubulin depends on NEDD1—and extended NEDD1 function to centriole duplication.\",\n      \"evidence\": \"siRNA depletion and GFP-fusion dominant-negative overexpression in human cells with nucleation/duplication readouts\",\n      \"pmids\": [\"16461362\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mapped binding only to C-terminal half, not a defined motif\", \"Mechanism linking NEDD1 to centriole duplication unresolved at this stage\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Distinguished centrosomal from spindle-based recruitment by showing FAM29A (Augmin component) tethers the NEDD1–γ-tubulin complex to spindle microtubules for MT-dependent amplification.\",\n      \"evidence\": \"Co-IP, mass spectrometry, siRNA depletion and nocodazole washout regrowth assays\",\n      \"pmids\": [\"19029337\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct NEDD1–Augmin interface not yet defined\", \"Did not address how partitioning is regulated\"]\n    },\n    {\n      \"year\": 2008,\n      \"claim\": \"Probed conservation of the targeting role across systems, finding NEDD1 in a non-γ-TuRC complex in Xenopus extracts and dispensable for centrosomal γ-tubulin targeting there, indicating context-dependent function.\",\n      \"evidence\": \"Immunodepletion and sucrose gradient fractionation of Xenopus egg extracts with nucleation assays\",\n      \"pmids\": [\"18252801\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Partially contradicts human-cell targeting data\", \"Basis of the species/system difference not resolved\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Showed how NEDD1 distribution between centrosome and spindle is controlled, placing Plk1 upstream of both FAM29A-dependent spindle targeting and direct centrosomal recruitment.\",\n      \"evidence\": \"Co-IP defining three separate complexes plus siRNA depletion and immunofluorescence in mammalian cells\",\n      \"pmids\": [\"19596795\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct Plk1 phosphosites on NEDD1 not mapped here\", \"Single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Defined the molecular basis of γ-tubulin binding by reconstituting a direct interaction through a 62-residue C-terminal helical domain that forms a tetramer.\",\n      \"evidence\": \"In vitro binding, mutagenesis, CD spectroscopy and analytical ultracentrifugation with cellular localization\",\n      \"pmids\": [\"20224777\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Quaternary arrangement within the intact γ-TuRC not visualized\", \"Single lab\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Extended NEDD1's role to acentriolar MTOCs, showing it is essential for meiotic spindle assembly and chromosome segregation fidelity in oocytes.\",\n      \"evidence\": \"siRNA knockdown in mouse oocytes with spindle, γ-tubulin, SAC and aneuploidy readouts\",\n      \"pmids\": [\"20079731\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Pericentrin-dependence shown by localization, not mechanism\", \"Single lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Identified the kinase and site (Nek9–Ser377) coupling Plk1 signaling to centrosomal NEDD1/γ-tubulin recruitment, separating it from the Nek6/Nek7 branch.\",\n      \"evidence\": \"In vitro kinase assay, phospho-specific antibodies, mutagenesis, Xenopus extracts and mammalian-cell depletion\",\n      \"pmids\": [\"22818914\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Ser377 phosphorylation mechanistically enhances centrosome binding unclear\", \"Single lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Showed that a cluster of phosphosites near the γ-tubulin-binding domain fine-tunes the NEDD1–γ-tubulin interaction and that CEP192 modulates this mitotic phosphorylation.\",\n      \"evidence\": \"MS phosphosite mapping, serine-to-alanine mutagenesis, Co-IP and siRNA rescue in human cells\",\n      \"pmids\": [\"22595525\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Responsible kinase(s) for the S557–S574 cluster not assigned\", \"Single lab\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined a spatially restricted nucleation pathway by showing Aurora A–Ser405 phosphorylation is specifically required for chromosome-proximal, RanGTP-driven nucleation but not centrosomal nucleation.\",\n      \"evidence\": \"In vitro kinase assay, phospho-specific antibodies, mutagenesis, Xenopus extract and human-cell rescue\",\n      \"pmids\": [\"23273898\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How Ser405 selectively licenses non-centrosomal nucleation unresolved\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Linked NEDD1 to PCM integrity by showing it recruits Cep57 to centrosomes, whose loss fragments the PCM and weakens nucleation.\",\n      \"evidence\": \"Co-IP and siRNA depletion with immunofluorescence in mammalian cells\",\n      \"pmids\": [\"22508265\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No reciprocal IP reported\", \"Direct binding interface undefined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Extended the Aurora A–Ser405 axis to cytokinesis, showing this phosphorylation concentrates NEDD1 at the midzone for central spindle microtubule nucleation.\",\n      \"evidence\": \"Aurora A inhibition with phosphomimetic/phospho-null NEDD1 rescue in human cells\",\n      \"pmids\": [\"31028180\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Midzone recruitment partner not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connected NEDD1 phosphorylation to centriole biogenesis by showing PLK4–Ser325 phosphorylation drives SAS-6 binding and cartwheel initiation.\",\n      \"evidence\": \"In vitro kinase assay, Co-IP, phosphomimetic/phospho-null mutagenesis and overexpression in human cells\",\n      \"pmids\": [\"33351100\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Structural basis of phospho-NEDD1–SAS-6 binding not determined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Directly visualized that Ser411 phosphorylation is required for branched microtubule nucleation and balances centrosome- versus chromosome-driven nucleation in spindle assembly.\",\n      \"evidence\": \"Phospho-variant stable HeLa lines and TIRF microscopy of MT branching in Xenopus egg extracts\",\n      \"pmids\": [\"36318115\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Responsible kinase for Ser411 not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Provided the structural model for how NEDD1 docks onto the γ-TuRC, showing a C-terminal tetrameric helical bundle in the cone lumen anchored via MZT1–GCP3 to GCP4/5/6 with WD40 domains projecting outward.\",\n      \"evidence\": \"Cryo-EM, AlphaFold modeling and biochemical pulldowns of NEDD1 mutants (preprint, published JCB 2025)\",\n      \"pmids\": [\"39574704\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How phosphorylation modulates this docked state structurally unaddressed\", \"Co-occupancy with CDK5RAP2 functional consequence unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Defined the WD40-domain interaction with Augmin at atomic resolution, showing it binds inside the Augmin V-junction and enhances dimerization to build a branching nucleation platform.\",\n      \"evidence\": \"Reconstitution of plant Augmin–NEDD1 complex, cryo-EM, crosslinking MS and evolutionary covariation\",\n      \"pmids\": [\"41387433\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Demonstrated with plant Augmin; full human complex not reconstituted\", \"Interplay with branching-specific phosphosites not tested\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identified a post-translational stability control whereby MZT1 suppresses NEDD1 ubiquitination, linking NEDD1 abundance to cancer cell proliferation and metabolic stress sensitivity.\",\n      \"evidence\": \"Ubiquitination assay, Co-IP, siRNA knockdown, Western blot and xenograft in gastric cancer cells\",\n      \"pmids\": [\"40204068\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"E3 ligase mediating NEDD1 ubiquitination not identified\", \"Single lab\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Extended NEDD1's microtubule-organizing role to differentiated cilia, localizing it to the basal foot of basal bodies where it is uniquely required for basal foot microtubule organization.\",\n      \"evidence\": \"Expansion microscopy 3D mapping and siRNA depletion in human airway multiciliated cells (preprint)\",\n      \"pmids\": [\"bio_10.1101_2025.09.04.674302\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Preprint; not peer-reviewed\", \"Recruitment mechanism to basal foot undefined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the distinct phosphorylation events (Ser325, Ser377, Ser405, Ser411 and the S557–S574 cluster) are spatially and temporally integrated on a single NEDD1 molecule to switch between centrosomal, chromosomal, midzone, branching and centriolar nucleation pathways remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No single study resolves combinatorial phospho-state of NEDD1 in vivo\", \"Kinases for several sites (S411, S557–S574 cluster) not all assigned\", \"Structural effect of phosphorylation on γ-TuRC/Augmin docking not determined\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 1, 5, 14, 16]},\n      {\"term_id\": \"GO:0008092\", \"supporting_discovery_ids\": [5, 14, 16]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 1, 7, 10]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [0, 2, 13]},\n      {\"term_id\": \"GO:0005929\", \"supporting_discovery_ids\": [18]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1, 6, 9, 12]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [1, 12, 18]}\n    ],\n    \"complexes\": [\n      \"γ-tubulin ring complex (γ-TuRC)\",\n      \"Augmin/HAUS complex\"\n    ],\n    \"partners\": [\n      \"TUBG1\",\n      \"FAM29A/HAUS6\",\n      \"MZT1\",\n      \"CEP192\",\n      \"CEP57\",\n      \"SAS-6\",\n      \"GCP4/TUBGCP4\",\n      \"GCP6/TUBGCP6\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}