{"gene":"NUDCD2","run_date":"2026-06-10T05:19:52","timeline":{"discoveries":[{"year":2010,"finding":"NudCL2 (NUDCD2) localizes to centrosomes in interphase and to spindle poles and kinetochores during mitosis. Depletion of NudCL2 destabilizes LIS1 protein and produces phenotypes resembling dynein or LIS1 deficiency. NudCL2 forms a complex with LIS1 and Hsp90, enhancing the LIS1–Hsp90 interaction; disruption of this interaction (via NudCL2 C-terminus overexpression) or inhibition of Hsp90 (geldanamycin) reduces LIS1 stability. Thus NudCL2 stabilizes LIS1 through the Hsp90 chaperone pathway.","method":"Immunofluorescence localization, co-immunoprecipitation, RNAi knockdown, western blotting, geldanamycin Hsp90 inhibition, C-terminus dominant-negative construct","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal co-IP, dominant-negative disruption, chemical inhibition, and knockdown phenotype with defined molecular readout in a single focused study; independently contextualized by review paper PMID:26965524","pmids":["20133715"],"is_preprint":false},{"year":2011,"finding":"Crystal and NMR structures of NudCL2 (mouse) show it shares a CS/p23 domain with other NudC family members but lacks the N-terminal coiled-coil dimerization motif; instead it dimerizes via unique domain swapping. In contrast to NudC and NudCL, NudCL2 does NOT inhibit aggregation of target proteins in vitro, indicating it lacks Hsp90-independent small heat-shock-protein chaperone activity. Additionally, none of the three NudC proteins form binary complexes with Lis1 in vitro.","method":"X-ray crystallography, NMR spectroscopy, in vitro aggregation chaperone assays, pull-down/binding assays","journal":"Journal of molecular biology","confidence":"High","confidence_rationale":"Tier 1 / Moderate — crystal structure and NMR plus functional in vitro chaperone assay in one study; negative binary Lis1 binding result is orthogonal to PMID:20133715 (complex requires cellular context/Hsp90)","pmids":["21530541"],"is_preprint":false},{"year":2018,"finding":"NudCL2 acts as an Hsp90 cochaperone that stabilizes cohesin subunits (SMC1, SMC3, RAD21, SA1/SA2) in mammalian cells. NudCL2 directly binds Hsp90 and stimulates its ATPase activity, enhancing Hsp90 chaperone function. Depletion of NudCL2 causes premature sister chromatid separation and mitotic defects; ectopic Hsp90 overexpression rescues cohesin instability caused by NudCL2 loss, but NudCL2 overexpression does not rescue Hsp90-inhibition phenotypes, placing NudCL2 upstream of Hsp90 in this pathway.","method":"Co-immunoprecipitation, western blotting, RNAi knockdown, Hsp90 ATPase activity assay, epistasis by rescue experiments (Hsp90 overexpression vs NudCL2 overexpression), mitotic chromosome spread analysis","journal":"Cellular and molecular life sciences : CMLS","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, in vitro ATPase assay, genetic epistasis by rescue, multiple orthogonal readouts in one focused study","pmids":["30368549"],"is_preprint":false},{"year":2019,"finding":"NudCL2 localizes to centrosomes and is required for accurate centriole duplication. Knockout or knockdown of NudCL2 causes centriole amplification; overexpression suppresses hydroxyurea-induced over-duplication. NudCL2 interacts with and stabilizes the E3 ubiquitin ligase HERC2. Loss of HERC2 phenocopies NudCL2 depletion and leads to accumulation of the positive centriole-duplication regulator USP33; knockdown of USP33 reverses centriole amplification in both NudCL2 KO and HERC2-depleted cells. Ectopic HERC2 rescues NudCL2-KO centriole amplification; NudCL2 overexpression does not rescue HERC2-depletion phenotype, defining the NudCL2→HERC2→USP33 pathway.","method":"CRISPR/Cas9 KO, RNAi, quantitative proteomics (iTRAQ), co-immunoprecipitation, western blotting, epistasis rescue experiments, immunofluorescence","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 2 / Strong — CRISPR KO, quantitative proteomics, Co-IP, and genetic epistasis with multiple rescue/depletion combinations in one focused study","pmids":["31427565"],"is_preprint":false},{"year":2020,"finding":"NudCL2 regulates single-cell (but not collective) migration by stabilizing both myosin-9 (MYH9) and LIS1 via Hsp90. NudCL2 binds myosin-9 (identified by Co-IP-mass spectrometry). NudCL2 depletion reduces myosin-9 levels and causes actin disorganization; ectopic myosin-9 rescues actin and single-cell migration defects. LIS1 depletion suppresses both single and collective migration (opposite to myosin-9 depletion); co-depletion of myosin-9 and LIS1 phenocopies NudCL2 depletion. Hsp90 inhibition also reduces myosin-9 stability; Hsp90 overexpression rescues myosin-9 instability caused by NudCL2 depletion.","method":"Co-immunoprecipitation coupled to mass spectrometry, RNAi knockdown, CRISPR KO, western blotting, actin staining, migration assays (single-cell and scratch wound), rescue overexpression experiments","journal":"Cell death & disease","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP-MS identification plus genetic epistasis via multiple KD/rescue combinations in one focused study","pmids":["32665550"],"is_preprint":false},{"year":2021,"finding":"NudCL2 functions as a selective autophagy receptor at mother centrioles, containing an LIR (LC3-interacting region) motif that bridges CP110 to the autophagosome marker LC3, thereby promoting autophagic degradation of CP110 and enabling ciliogenesis initiation. NudCL2 KO prevents CP110 removal from mother centrioles and blocks ciliogenesis; wild-type but not LIR-mutant NudCL2 rescues these defects. CP110 knockdown attenuates ciliogenesis defects in NudCL2-deficient cells. In zebrafish, NudCL2 morphants show ciliation-related phenotypes reversed by wild-type but not LIR-mutant NudCL2, and by CP110 depletion.","method":"CRISPR/Cas9 KO, RNAi, co-immunoprecipitation (NudCL2–LC3, NudCL2–CP110), LIR-motif mutagenesis, immunofluorescence, zebrafish morpholino knockdown, rescue experiments","journal":"Cell research","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal Co-IP, LIR mutagenesis, CRISPR KO + rescue, in vivo zebrafish validation; multiple orthogonal methods in one focused study","pmids":["34480124"],"is_preprint":false},{"year":2024,"finding":"NudCL2 localizes to the midbody and is required for cytokinesis. Its depletion causes cytokinesis failure, multinucleation, and midbody disorganization. iTRAQ quantitative proteomics identified RCC2 as downregulated in NudCL2 KO cells. NudCL2 interacts with RCC2, and Hsp90 forms a complex with NudCL2 to stabilize RCC2. RCC2 depletion phenocopies NudCL2 loss; ectopic RCC2 rescues NudCL2-KO cytokinesis defects, but NudCL2 overexpression does not rescue RCC2-depletion defects, establishing a NudCL2/Hsp90/RCC2 pathway for cytokinesis.","method":"CRISPR/Cas9 KO, RNAi, iTRAQ quantitative proteomics, co-immunoprecipitation, western blotting, immunofluorescence (midbody localization), epistasis rescue experiments","journal":"Protein & cell","confidence":"High","confidence_rationale":"Tier 2 / Strong — quantitative proteomics, Co-IP, CRISPR KO, genetic epistasis with multiple rescue/depletion combinations in one focused study","pmids":["38801297"],"is_preprint":false},{"year":2024,"finding":"RetSat interacts with cohesin/condensin components SMC1A and NudCL2 (NUDCD2) in embryonic stem cells; RetSat deletion impairs chromosome loading of SMC1A, SMC3, and NudCL2, linking NudCL2 to chromosome condensation/cohesion in pluripotent cells.","method":"Co-immunoprecipitation, LC-MS/MS, immunoblotting, immunostaining, RetSat KO mouse ESCs","journal":"Cellular and molecular life sciences : CMLS","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — Co-IP and LC-MS/MS in a single study where NudCL2 is a secondary finding; no direct functional manipulation of NudCL2 itself","pmids":["39172275"],"is_preprint":false},{"year":2025,"finding":"NudCL2 suppresses pancreatic cancer cell invasion and metastasis by regulating the transcriptional activity of SLC7A11; NudCL2 depletion upregulates SLC7A11, activating the EMT pathway. SLC7A11 knockdown reverses the increased motility caused by NudCL2 loss, placing SLC7A11 downstream of NudCL2 in EMT regulation.","method":"RNAi knockdown, invasion/migration assays, in vivo lung and liver metastasis models, western blotting, transcriptional activity assay, epistasis (SLC7A11 KD reversal of NudCL2-KD phenotype)","journal":"Experimental cell research","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — genetic epistasis and functional assays in a single study; transcriptional regulation mechanism not fully resolved at molecular level","pmids":["40907782"],"is_preprint":false}],"current_model":"NUDCD2 (NudCL2) is an Hsp90 cochaperone that stabilizes multiple client proteins—including LIS1, cohesin subunits, myosin-9, HERC2, and RCC2—by enhancing Hsp90 ATPase/chaperone activity; it localizes to centrosomes, spindle poles, kinetochores, and the midbody to regulate cell division (sister chromatid cohesion, centriole duplication, cytokinesis), and at mother centrioles it acts as a selective autophagy receptor via its LIR motif to degrade CP110 and initiate ciliogenesis, while also modulating single-cell migration through balanced stabilization of myosin-9 and LIS1."},"narrative":{"mechanistic_narrative":"NUDCD2 (NudCL2) is an Hsp90 cochaperone that governs cell division and ciliogenesis by enforcing the stability of multiple client proteins at distinct mitotic and centriolar structures [PMID:20133715, PMID:30368549]. It binds Hsp90 directly and stimulates its ATPase activity, acting upstream of Hsp90 to enhance chaperone-dependent client folding; consistently, ectopic Hsp90 rescues client instability caused by NudCL2 loss while NudCL2 overexpression cannot rescue Hsp90 inhibition [PMID:30368549]. Through this mechanism it stabilizes LIS1 [PMID:20133715], the cohesin subunits SMC1/SMC3/RAD21/SA1-SA2 to prevent premature sister chromatid separation [PMID:30368549], myosin-9 to organize actin and drive single-cell migration [PMID:32665550], and RCC2 at the midbody to complete cytokinesis [PMID:38801297]. NudCL2 also controls centriole copy number through a NudCL2→HERC2→USP33 axis, stabilizing the E3 ligase HERC2 to limit accumulation of the centriole-duplication regulator USP33 [PMID:31427565]. At mother centrioles it switches roles to a selective autophagy receptor, using an LIR motif to bridge CP110 to LC3 and target CP110 for autophagic degradation, thereby licensing ciliogenesis as validated in zebrafish [PMID:34480124]. Structurally it carries a CS/p23 domain shared with the NudC family but dimerizes by domain swapping and lacks intrinsic Hsp90-independent anti-aggregation chaperone activity [PMID:21530541].","teleology":[{"year":2010,"claim":"Established NudCL2 as an Hsp90-pathway cochaperone by showing it stabilizes the dynein regulator LIS1, answering how LIS1 levels are maintained during mitosis.","evidence":"Immunofluorescence, reciprocal co-IP of NudCL2-LIS1-Hsp90, RNAi knockdown phenotypes, dominant-negative C-terminus, and geldanamycin Hsp90 inhibition in mammalian cells","pmids":["20133715"],"confidence":"High","gaps":["Did not resolve whether NudCL2 contacts LIS1 directly or only via Hsp90","Mechanism of how NudCL2 enhances Hsp90 activity not defined at this stage"]},{"year":2011,"claim":"Defined the structural basis of NudCL2, showing a CS/p23 domain with domain-swap dimerization and, critically, that it lacks intrinsic anti-aggregation activity and does not bind LIS1 in a binary complex in vitro.","evidence":"X-ray crystallography and NMR of mouse NudCL2 plus in vitro aggregation and pull-down assays","pmids":["21530541"],"confidence":"High","gaps":["In vitro absence of binary LIS1 binding leaves the cellular requirement for Hsp90/context unexplained","No structure of NudCL2 bound to Hsp90 or any client"]},{"year":2018,"claim":"Placed NudCL2 mechanistically upstream of Hsp90 and broadened its client repertoire to cohesin, explaining how sister chromatid cohesion is safeguarded.","evidence":"Co-IP, in vitro Hsp90 ATPase assay, RNAi, chromosome spreads, and genetic epistasis by Hsp90 versus NudCL2 overexpression rescue","pmids":["30368549"],"confidence":"High","gaps":["Whether NudCL2 selects cohesin subunits directly or via a recognition motif unknown","Stoichiometry of the NudCL2-Hsp90-cohesin complex not determined"]},{"year":2019,"claim":"Connected NudCL2 to centriole copy-number control by defining a NudCL2→HERC2→USP33 pathway, answering how it prevents centriole amplification.","evidence":"CRISPR KO, iTRAQ proteomics, co-IP, immunofluorescence, and epistasis with HERC2 and USP33 depletion/rescue","pmids":["31427565"],"confidence":"High","gaps":["Whether HERC2 stabilization uses the canonical Hsp90 mechanism not directly tested","How HERC2 controls USP33 levels biochemically unresolved"]},{"year":2020,"claim":"Extended NudCL2 client stabilization to myosin-9 and explained selective control of single-cell versus collective migration through opposing myosin-9 and LIS1 effects.","evidence":"Co-IP-mass spectrometry, RNAi/CRISPR, actin staining, single-cell and wound migration assays, and Hsp90-dependent rescue experiments","pmids":["32665550"],"confidence":"High","gaps":["Molecular basis for the divergent migration outcomes of two clients not fully mechanized","Direct versus Hsp90-mediated myosin-9 binding not separated"]},{"year":2024,"claim":"Defined a NudCL2/Hsp90/RCC2 pathway at the midbody required to complete cytokinesis, extending its division roles to the final abscission step.","evidence":"CRISPR KO, iTRAQ proteomics, co-IP, midbody immunofluorescence, and epistasis with RCC2 depletion/rescue","pmids":["38801297"],"confidence":"High","gaps":["How NudCL2 is targeted to the midbody not defined","Direct RCC2 recognition determinants unknown"]},{"year":2021,"claim":"Revealed a chaperone-independent role for NudCL2 as a selective autophagy receptor that uses an LIR motif to degrade CP110 and initiate ciliogenesis.","evidence":"CRISPR KO, reciprocal co-IP (NudCL2-LC3 and NudCL2-CP110), LIR mutagenesis with rescue, and zebrafish morpholino in vivo validation","pmids":["34480124"],"confidence":"High","gaps":["How NudCL2 is recruited specifically to mother centrioles for this function unknown","Relationship between the autophagy-receptor and Hsp90-cochaperone roles not integrated"]},{"year":2024,"claim":"Linked NudCL2 to chromosome loading of cohesin/condensin components in pluripotent cells, suggesting a role beyond protein stabilization in chromosome organization.","evidence":"Co-IP, LC-MS/MS, immunostaining in RetSat-knockout mouse embryonic stem cells (NudCL2 as secondary finding)","pmids":["39172275"],"confidence":"Medium","gaps":["No direct functional manipulation of NudCL2 in this system","Whether NudCL2 chromosome loading is cause or consequence of cohesin recruitment unclear"]},{"year":2025,"claim":"Implicated NudCL2 as a suppressor of pancreatic cancer invasion via transcriptional control of SLC7A11 and the EMT pathway, extending its function to metastasis regulation.","evidence":"RNAi, invasion/migration assays, in vivo metastasis models, and epistasis showing SLC7A11 knockdown reverses NudCL2-loss phenotype","pmids":["40907782"],"confidence":"Medium","gaps":["Molecular mechanism by which NudCL2 regulates SLC7A11 transcription not resolved","Connection between this transcriptional role and the cochaperone function unestablished"]},{"year":null,"claim":"It remains unknown how NudCL2 achieves client selectivity across its diverse substrates and how its Hsp90-cochaperone activity is mechanistically reconciled with its LIR-dependent autophagy-receptor and transcriptional-regulatory roles.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No structure of NudCL2 bound to Hsp90 or any client","No unifying determinant of substrate recognition identified","Switch between chaperone and autophagy-receptor modes uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0140096","term_label":"catalytic activity, acting on a protein","supporting_discovery_ids":[0,2]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[0,2,3,4,6]},{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[5]},{"term_id":"GO:0044183","term_label":"protein folding chaperone","supporting_discovery_ids":[0,2]}],"localization":[{"term_id":"GO:0005815","term_label":"microtubule organizing center","supporting_discovery_ids":[0,3]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0]},{"term_id":"GO:0005856","term_label":"cytoskeleton","supporting_discovery_ids":[4]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,2,6]},{"term_id":"R-HSA-392499","term_label":"Metabolism of proteins","supporting_discovery_ids":[0,2,4,6]},{"term_id":"R-HSA-9612973","term_label":"Autophagy","supporting_discovery_ids":[5]},{"term_id":"R-HSA-1852241","term_label":"Organelle biogenesis and maintenance","supporting_discovery_ids":[3,5]}],"complexes":[],"partners":["HSP90","LIS1","MYH9","HERC2","RCC2","LC3","CP110","SMC1A"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8WVJ2","full_name":"NudC domain-containing protein 2","aliases":[],"length_aa":157,"mass_kda":17.7,"function":"May regulate the LIS1/dynein pathway by stabilizing LIS1 with Hsp90 chaperone","subcellular_location":"Chromosome, centromere, kinetochore; Cytoplasm, cytoskeleton, microtubule organizing center, centrosome; Cytoplasm, cytoskeleton, spindle pole","url":"https://www.uniprot.org/uniprotkb/Q8WVJ2/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":false,"resolved_as":"","url":"https://depmap.org/portal/gene/NUDCD2","classification":"Not Classified","n_dependent_lines":46,"n_total_lines":1208,"dependency_fraction":0.0380794701986755},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"DDOST","stoichiometry":0.2},{"gene":"FKBP5","stoichiometry":0.2},{"gene":"OST4","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/NUDCD2","total_profiled":1310},"omim":[{"mim_id":"620136","title":"NUDC DOMAIN-CONTAINING PROTEIN 2; NUDCD2","url":"https://www.omim.org/entry/620136"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Microtubules","reliability":"Supported"},{"location":"Cytokinetic bridge","reliability":"Additional"},{"location":"Mitotic spindle","reliability":"Additional"},{"location":"Cytosol","reliability":"Additional"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/NUDCD2"},"hgnc":{"alias_symbol":["DKFZp686E10109","NudCL2"],"prev_symbol":[]},"alphafold":{"accession":"Q8WVJ2","domains":[{"cath_id":"2.60.40.790","chopping":"4-129","consensus_level":"medium","plddt":88.5484,"start":4,"end":129}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8WVJ2","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8WVJ2-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8WVJ2-F1-predicted_aligned_error_v6.png","plddt_mean":83.5},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=NUDCD2","jax_strain_url":"https://www.jax.org/strain/search?query=NUDCD2"},"sequence":{"accession":"Q8WVJ2","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8WVJ2.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8WVJ2/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8WVJ2"}},"corpus_meta":[{"pmid":"22781929","id":"PMC_22781929","title":"Transcriptional response of the bovine endometrium and embryo to endometrial polymorphonuclear neutrophil infiltration as an indicator of subclinical inflammation of the uterine environment.","date":"2012","source":"Reproduction, fertility, and development","url":"https://pubmed.ncbi.nlm.nih.gov/22781929","citation_count":38,"is_preprint":false},{"pmid":"26965524","id":"PMC_26965524","title":"Emerging roles of NudC family: from molecular regulation to clinical implications.","date":"2016","source":"Science China. 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Depletion of NudCL2 destabilizes LIS1 protein and produces phenotypes resembling dynein or LIS1 deficiency. NudCL2 forms a complex with LIS1 and Hsp90, enhancing the LIS1–Hsp90 interaction; disruption of this interaction (via NudCL2 C-terminus overexpression) or inhibition of Hsp90 (geldanamycin) reduces LIS1 stability. Thus NudCL2 stabilizes LIS1 through the Hsp90 chaperone pathway.\",\n      \"method\": \"Immunofluorescence localization, co-immunoprecipitation, RNAi knockdown, western blotting, geldanamycin Hsp90 inhibition, C-terminus dominant-negative construct\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal co-IP, dominant-negative disruption, chemical inhibition, and knockdown phenotype with defined molecular readout in a single focused study; independently contextualized by review paper PMID:26965524\",\n      \"pmids\": [\"20133715\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"Crystal and NMR structures of NudCL2 (mouse) show it shares a CS/p23 domain with other NudC family members but lacks the N-terminal coiled-coil dimerization motif; instead it dimerizes via unique domain swapping. In contrast to NudC and NudCL, NudCL2 does NOT inhibit aggregation of target proteins in vitro, indicating it lacks Hsp90-independent small heat-shock-protein chaperone activity. Additionally, none of the three NudC proteins form binary complexes with Lis1 in vitro.\",\n      \"method\": \"X-ray crystallography, NMR spectroscopy, in vitro aggregation chaperone assays, pull-down/binding assays\",\n      \"journal\": \"Journal of molecular biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 / Moderate — crystal structure and NMR plus functional in vitro chaperone assay in one study; negative binary Lis1 binding result is orthogonal to PMID:20133715 (complex requires cellular context/Hsp90)\",\n      \"pmids\": [\"21530541\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"NudCL2 acts as an Hsp90 cochaperone that stabilizes cohesin subunits (SMC1, SMC3, RAD21, SA1/SA2) in mammalian cells. NudCL2 directly binds Hsp90 and stimulates its ATPase activity, enhancing Hsp90 chaperone function. Depletion of NudCL2 causes premature sister chromatid separation and mitotic defects; ectopic Hsp90 overexpression rescues cohesin instability caused by NudCL2 loss, but NudCL2 overexpression does not rescue Hsp90-inhibition phenotypes, placing NudCL2 upstream of Hsp90 in this pathway.\",\n      \"method\": \"Co-immunoprecipitation, western blotting, RNAi knockdown, Hsp90 ATPase activity assay, epistasis by rescue experiments (Hsp90 overexpression vs NudCL2 overexpression), mitotic chromosome spread analysis\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, in vitro ATPase assay, genetic epistasis by rescue, multiple orthogonal readouts in one focused study\",\n      \"pmids\": [\"30368549\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"NudCL2 localizes to centrosomes and is required for accurate centriole duplication. Knockout or knockdown of NudCL2 causes centriole amplification; overexpression suppresses hydroxyurea-induced over-duplication. NudCL2 interacts with and stabilizes the E3 ubiquitin ligase HERC2. Loss of HERC2 phenocopies NudCL2 depletion and leads to accumulation of the positive centriole-duplication regulator USP33; knockdown of USP33 reverses centriole amplification in both NudCL2 KO and HERC2-depleted cells. Ectopic HERC2 rescues NudCL2-KO centriole amplification; NudCL2 overexpression does not rescue HERC2-depletion phenotype, defining the NudCL2→HERC2→USP33 pathway.\",\n      \"method\": \"CRISPR/Cas9 KO, RNAi, quantitative proteomics (iTRAQ), co-immunoprecipitation, western blotting, epistasis rescue experiments, immunofluorescence\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — CRISPR KO, quantitative proteomics, Co-IP, and genetic epistasis with multiple rescue/depletion combinations in one focused study\",\n      \"pmids\": [\"31427565\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"NudCL2 regulates single-cell (but not collective) migration by stabilizing both myosin-9 (MYH9) and LIS1 via Hsp90. NudCL2 binds myosin-9 (identified by Co-IP-mass spectrometry). NudCL2 depletion reduces myosin-9 levels and causes actin disorganization; ectopic myosin-9 rescues actin and single-cell migration defects. LIS1 depletion suppresses both single and collective migration (opposite to myosin-9 depletion); co-depletion of myosin-9 and LIS1 phenocopies NudCL2 depletion. Hsp90 inhibition also reduces myosin-9 stability; Hsp90 overexpression rescues myosin-9 instability caused by NudCL2 depletion.\",\n      \"method\": \"Co-immunoprecipitation coupled to mass spectrometry, RNAi knockdown, CRISPR KO, western blotting, actin staining, migration assays (single-cell and scratch wound), rescue overexpression experiments\",\n      \"journal\": \"Cell death & disease\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP-MS identification plus genetic epistasis via multiple KD/rescue combinations in one focused study\",\n      \"pmids\": [\"32665550\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"NudCL2 functions as a selective autophagy receptor at mother centrioles, containing an LIR (LC3-interacting region) motif that bridges CP110 to the autophagosome marker LC3, thereby promoting autophagic degradation of CP110 and enabling ciliogenesis initiation. NudCL2 KO prevents CP110 removal from mother centrioles and blocks ciliogenesis; wild-type but not LIR-mutant NudCL2 rescues these defects. CP110 knockdown attenuates ciliogenesis defects in NudCL2-deficient cells. In zebrafish, NudCL2 morphants show ciliation-related phenotypes reversed by wild-type but not LIR-mutant NudCL2, and by CP110 depletion.\",\n      \"method\": \"CRISPR/Cas9 KO, RNAi, co-immunoprecipitation (NudCL2–LC3, NudCL2–CP110), LIR-motif mutagenesis, immunofluorescence, zebrafish morpholino knockdown, rescue experiments\",\n      \"journal\": \"Cell research\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal Co-IP, LIR mutagenesis, CRISPR KO + rescue, in vivo zebrafish validation; multiple orthogonal methods in one focused study\",\n      \"pmids\": [\"34480124\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"NudCL2 localizes to the midbody and is required for cytokinesis. Its depletion causes cytokinesis failure, multinucleation, and midbody disorganization. iTRAQ quantitative proteomics identified RCC2 as downregulated in NudCL2 KO cells. NudCL2 interacts with RCC2, and Hsp90 forms a complex with NudCL2 to stabilize RCC2. RCC2 depletion phenocopies NudCL2 loss; ectopic RCC2 rescues NudCL2-KO cytokinesis defects, but NudCL2 overexpression does not rescue RCC2-depletion defects, establishing a NudCL2/Hsp90/RCC2 pathway for cytokinesis.\",\n      \"method\": \"CRISPR/Cas9 KO, RNAi, iTRAQ quantitative proteomics, co-immunoprecipitation, western blotting, immunofluorescence (midbody localization), epistasis rescue experiments\",\n      \"journal\": \"Protein & cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — quantitative proteomics, Co-IP, CRISPR KO, genetic epistasis with multiple rescue/depletion combinations in one focused study\",\n      \"pmids\": [\"38801297\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"RetSat interacts with cohesin/condensin components SMC1A and NudCL2 (NUDCD2) in embryonic stem cells; RetSat deletion impairs chromosome loading of SMC1A, SMC3, and NudCL2, linking NudCL2 to chromosome condensation/cohesion in pluripotent cells.\",\n      \"method\": \"Co-immunoprecipitation, LC-MS/MS, immunoblotting, immunostaining, RetSat KO mouse ESCs\",\n      \"journal\": \"Cellular and molecular life sciences : CMLS\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — Co-IP and LC-MS/MS in a single study where NudCL2 is a secondary finding; no direct functional manipulation of NudCL2 itself\",\n      \"pmids\": [\"39172275\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"NudCL2 suppresses pancreatic cancer cell invasion and metastasis by regulating the transcriptional activity of SLC7A11; NudCL2 depletion upregulates SLC7A11, activating the EMT pathway. SLC7A11 knockdown reverses the increased motility caused by NudCL2 loss, placing SLC7A11 downstream of NudCL2 in EMT regulation.\",\n      \"method\": \"RNAi knockdown, invasion/migration assays, in vivo lung and liver metastasis models, western blotting, transcriptional activity assay, epistasis (SLC7A11 KD reversal of NudCL2-KD phenotype)\",\n      \"journal\": \"Experimental cell research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — genetic epistasis and functional assays in a single study; transcriptional regulation mechanism not fully resolved at molecular level\",\n      \"pmids\": [\"40907782\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"NUDCD2 (NudCL2) is an Hsp90 cochaperone that stabilizes multiple client proteins—including LIS1, cohesin subunits, myosin-9, HERC2, and RCC2—by enhancing Hsp90 ATPase/chaperone activity; it localizes to centrosomes, spindle poles, kinetochores, and the midbody to regulate cell division (sister chromatid cohesion, centriole duplication, cytokinesis), and at mother centrioles it acts as a selective autophagy receptor via its LIR motif to degrade CP110 and initiate ciliogenesis, while also modulating single-cell migration through balanced stabilization of myosin-9 and LIS1.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"NUDCD2 (NudCL2) is an Hsp90 cochaperone that governs cell division and ciliogenesis by enforcing the stability of multiple client proteins at distinct mitotic and centriolar structures [#0, #2]. It binds Hsp90 directly and stimulates its ATPase activity, acting upstream of Hsp90 to enhance chaperone-dependent client folding; consistently, ectopic Hsp90 rescues client instability caused by NudCL2 loss while NudCL2 overexpression cannot rescue Hsp90 inhibition [#2]. Through this mechanism it stabilizes LIS1 [#0], the cohesin subunits SMC1/SMC3/RAD21/SA1-SA2 to prevent premature sister chromatid separation [#2], myosin-9 to organize actin and drive single-cell migration [#4], and RCC2 at the midbody to complete cytokinesis [#6]. NudCL2 also controls centriole copy number through a NudCL2\\u2192HERC2\\u2192USP33 axis, stabilizing the E3 ligase HERC2 to limit accumulation of the centriole-duplication regulator USP33 [#3]. At mother centrioles it switches roles to a selective autophagy receptor, using an LIR motif to bridge CP110 to LC3 and target CP110 for autophagic degradation, thereby licensing ciliogenesis as validated in zebrafish [#5]. Structurally it carries a CS/p23 domain shared with the NudC family but dimerizes by domain swapping and lacks intrinsic Hsp90-independent anti-aggregation chaperone activity [#1].\",\n  \"teleology\": [\n    {\n      \"year\": 2010,\n      \"claim\": \"Established NudCL2 as an Hsp90-pathway cochaperone by showing it stabilizes the dynein regulator LIS1, answering how LIS1 levels are maintained during mitosis.\",\n      \"evidence\": \"Immunofluorescence, reciprocal co-IP of NudCL2-LIS1-Hsp90, RNAi knockdown phenotypes, dominant-negative C-terminus, and geldanamycin Hsp90 inhibition in mammalian cells\",\n      \"pmids\": [\"20133715\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve whether NudCL2 contacts LIS1 directly or only via Hsp90\", \"Mechanism of how NudCL2 enhances Hsp90 activity not defined at this stage\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Defined the structural basis of NudCL2, showing a CS/p23 domain with domain-swap dimerization and, critically, that it lacks intrinsic anti-aggregation activity and does not bind LIS1 in a binary complex in vitro.\",\n      \"evidence\": \"X-ray crystallography and NMR of mouse NudCL2 plus in vitro aggregation and pull-down assays\",\n      \"pmids\": [\"21530541\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"In vitro absence of binary LIS1 binding leaves the cellular requirement for Hsp90/context unexplained\", \"No structure of NudCL2 bound to Hsp90 or any client\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Placed NudCL2 mechanistically upstream of Hsp90 and broadened its client repertoire to cohesin, explaining how sister chromatid cohesion is safeguarded.\",\n      \"evidence\": \"Co-IP, in vitro Hsp90 ATPase assay, RNAi, chromosome spreads, and genetic epistasis by Hsp90 versus NudCL2 overexpression rescue\",\n      \"pmids\": [\"30368549\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether NudCL2 selects cohesin subunits directly or via a recognition motif unknown\", \"Stoichiometry of the NudCL2-Hsp90-cohesin complex not determined\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Connected NudCL2 to centriole copy-number control by defining a NudCL2\\u2192HERC2\\u2192USP33 pathway, answering how it prevents centriole amplification.\",\n      \"evidence\": \"CRISPR KO, iTRAQ proteomics, co-IP, immunofluorescence, and epistasis with HERC2 and USP33 depletion/rescue\",\n      \"pmids\": [\"31427565\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether HERC2 stabilization uses the canonical Hsp90 mechanism not directly tested\", \"How HERC2 controls USP33 levels biochemically unresolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Extended NudCL2 client stabilization to myosin-9 and explained selective control of single-cell versus collective migration through opposing myosin-9 and LIS1 effects.\",\n      \"evidence\": \"Co-IP-mass spectrometry, RNAi/CRISPR, actin staining, single-cell and wound migration assays, and Hsp90-dependent rescue experiments\",\n      \"pmids\": [\"32665550\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Molecular basis for the divergent migration outcomes of two clients not fully mechanized\", \"Direct versus Hsp90-mediated myosin-9 binding not separated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Defined a NudCL2/Hsp90/RCC2 pathway at the midbody required to complete cytokinesis, extending its division roles to the final abscission step.\",\n      \"evidence\": \"CRISPR KO, iTRAQ proteomics, co-IP, midbody immunofluorescence, and epistasis with RCC2 depletion/rescue\",\n      \"pmids\": [\"38801297\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How NudCL2 is targeted to the midbody not defined\", \"Direct RCC2 recognition determinants unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Revealed a chaperone-independent role for NudCL2 as a selective autophagy receptor that uses an LIR motif to degrade CP110 and initiate ciliogenesis.\",\n      \"evidence\": \"CRISPR KO, reciprocal co-IP (NudCL2-LC3 and NudCL2-CP110), LIR mutagenesis with rescue, and zebrafish morpholino in vivo validation\",\n      \"pmids\": [\"34480124\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How NudCL2 is recruited specifically to mother centrioles for this function unknown\", \"Relationship between the autophagy-receptor and Hsp90-cochaperone roles not integrated\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Linked NudCL2 to chromosome loading of cohesin/condensin components in pluripotent cells, suggesting a role beyond protein stabilization in chromosome organization.\",\n      \"evidence\": \"Co-IP, LC-MS/MS, immunostaining in RetSat-knockout mouse embryonic stem cells (NudCL2 as secondary finding)\",\n      \"pmids\": [\"39172275\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No direct functional manipulation of NudCL2 in this system\", \"Whether NudCL2 chromosome loading is cause or consequence of cohesin recruitment unclear\"]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Implicated NudCL2 as a suppressor of pancreatic cancer invasion via transcriptional control of SLC7A11 and the EMT pathway, extending its function to metastasis regulation.\",\n      \"evidence\": \"RNAi, invasion/migration assays, in vivo metastasis models, and epistasis showing SLC7A11 knockdown reverses NudCL2-loss phenotype\",\n      \"pmids\": [\"40907782\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism by which NudCL2 regulates SLC7A11 transcription not resolved\", \"Connection between this transcriptional role and the cochaperone function unestablished\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unknown how NudCL2 achieves client selectivity across its diverse substrates and how its Hsp90-cochaperone activity is mechanistically reconciled with its LIR-dependent autophagy-receptor and transcriptional-regulatory roles.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No structure of NudCL2 bound to Hsp90 or any client\", \"No unifying determinant of substrate recognition identified\", \"Switch between chaperone and autophagy-receptor modes uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0140096\", \"supporting_discovery_ids\": [0, 2]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [0, 2, 3, 4, 6]},\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"GO:0044183\", \"supporting_discovery_ids\": [0, 2]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005815\", \"supporting_discovery_ids\": [0, 3]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0005856\", \"supporting_discovery_ids\": [4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 2, 6]},\n      {\"term_id\": \"R-HSA-392499\", \"supporting_discovery_ids\": [0, 2, 4, 6]},\n      {\"term_id\": \"R-HSA-9612973\", \"supporting_discovery_ids\": [5]},\n      {\"term_id\": \"R-HSA-1852241\", \"supporting_discovery_ids\": [3, 5]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"HSP90\", \"LIS1\", \"MYH9\", \"HERC2\", \"RCC2\", \"LC3\", \"CP110\", \"SMC1A\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}