{"gene":"CENPW","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2007,"finding":"CENPW (CUG2) protein localizes predominantly to the nucleus when expressed as EGFP fusion, and overexpression in NIH3T3 mouse fibroblasts induces cancer-specific phenotypes in vitro and tumor formation in nude mice, establishing proto-oncogenic activity.","method":"EGFP fusion protein localization imaging; soft-agar and nude mouse xenograft assays","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct localization experiment and functional KO/OE assay, single lab, single study","pmids":["17610844"],"is_preprint":false},{"year":2009,"finding":"CENPW (CUG2) physically interacts with CENP-T (a component of the CENP-A nucleosome complex) and CENP-A in a centromeric complex, co-localizes with centromeric markers, and is required for proper chromosome segregation during mitosis; its depletion induces aberrant cell division.","method":"Yeast two-hybrid screening; co-immunoprecipitation; immunofluorescent staining; siRNA knockdown with cell viability and division phenotype readout","journal":"Molecules and cells","confidence":"High","confidence_rationale":"Tier 2 / Strong — yeast two-hybrid plus Co-IP plus immunofluorescence plus functional knockdown, replicated across multiple subsequent studies","pmids":["19533040"],"is_preprint":false},{"year":2010,"finding":"The CENPW promoter contains a GC-rich Sp1-binding site (-46 to -36) that is critical for basal and serum-induced expression; Sp1 and Sp3 transcription factors specifically bind this site and mediate transactivation of CENPW.","method":"Promoter deletion analysis; competitive EMSA with mutated oligos; supershift assays with Sp1/Sp3 antibodies; serum stimulation experiments","journal":"Molecular biology reports","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — EMSA with mutagenesis and supershift, single lab, two orthogonal methods","pmids":["20180024"],"is_preprint":false},{"year":2010,"finding":"CUG2 expression activates MAPK (ERK, JNK, p38), Src kinase, and Ras signaling; inhibition of Ras or p38 MAPK (but not ERK, JNK, or Src) blocks reoviral replication in CUG2-expressing cells, establishing Ras and p38 as necessary downstream effectors of CUG2-mediated permissiveness to reovirus.","method":"Pharmacological inhibitors of specific kinases; stable CUG2-overexpressing NIH3T3 cells; viral replication assays","journal":"Cancer gene therapy","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — epistasis via selective inhibitors with clear pathway placement, single lab","pmids":["20075984"],"is_preprint":false},{"year":2011,"finding":"CENPW (CENP-W) localizes to the nucleolus and nuclear matrix, associates with both RNA and DNA by fractionation, and physically interacts with the nucleolar phosphoprotein nucleophosmin (B23/NPM1); depletion of B23 by siRNA decreases CENP-W protein stability and causes its severe mislocalization during prophase.","method":"Subnuclear fractionation; RNA/DNA association assays; biochemical affinity binding (co-IP); siRNA knockdown; immunofluorescence microscopy","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (fractionation, affinity binding, siRNA with localization phenotype) in single rigorous study","pmids":["22002061"],"is_preprint":false},{"year":2011,"finding":"In zebrafish, morpholino-mediated knockdown of cug2 causes mitotic arrest with abnormal spindle formation and chromosome misalignment in the neural plate, followed by CNS-wide apoptosis, establishing an in vivo requirement for Cug2 in normal mitosis during neurogenesis.","method":"Morpholino knockdown in zebrafish embryos; spindle/chromosome immunofluorescence; apoptosis assays","journal":"BMC developmental biology","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean in vivo loss-of-function with defined mitotic phenotype, single lab","pmids":["21838932"],"is_preprint":false},{"year":2013,"finding":"CSN5/JAB1 directly interacts with both CENP-T and CENP-W (identified by yeast two-hybrid and confirmed by Co-IP), promotes their ubiquitin- and proteasome-dependent degradation, and formation of the CENP-T·CENP-W complex stabilizes both proteins by blocking CSN5-mediated degradation; dysregulation of CSN5 causes severe defects in CENP-T·CENP-W recruitment to the kinetochore during prophase.","method":"Yeast two-hybrid; co-immunoprecipitation; ubiquitination/proteasome degradation assays; immunofluorescence of kinetochore recruitment","journal":"The Journal of biological chemistry","confidence":"High","confidence_rationale":"Tier 2 / Strong — yeast two-hybrid plus Co-IP plus functional degradation assays plus localization phenotype, multiple orthogonal methods in single study","pmids":["23926101"],"is_preprint":false},{"year":2014,"finding":"CENP-W depletion in HeLa cells causes bipolar spindle fragmentation into multipolar spindles, depletion of Hec1 at kinetochores, abnormal centriole splitting, and generation of acentriolar spindle poles; spindle pole fragmentation requires microtubules (absent in nocodazole) and Eg5 motor activity (reduced by monastrol), implicating CENP-W in maintaining kinetochore-microtubule attachment that resists motor-generated traction forces.","method":"RNAi depletion; live-cell fluorescence imaging (H2B and tubulin); immunofluorescence of centrioles and centrosomal markers; pharmacological manipulation (nocodazole, monastrol); TPX2 overexpression rescue","journal":"PloS one","confidence":"High","confidence_rationale":"Tier 2 / Strong — multiple orthogonal methods (RNAi, live imaging, pharmacological epistasis, genetic rescue) in single rigorous study","pmids":["25329824"],"is_preprint":false},{"year":2015,"finding":"CENP-W physically associates with EZH2 (catalytic subunit of PRC2), enhances EZH2 protein stability, and is recruited to the promoters of EZH2 target genes (by chromatin immunoprecipitation) to facilitate EZH2-mediated transcriptional repression (H3K27me3-associated gene silencing).","method":"Co-immunoprecipitation; protein stability assays; chromatin immunoprecipitation (ChIP)","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus ChIP, single lab, two orthogonal methods","pmids":["26111449"],"is_preprint":false},{"year":2016,"finding":"CENP-W physically interacts with hnRNP U; the interaction mutually stabilizes both proteins by inhibiting proteasome-mediated degradation. They co-localize in the nuclear matrix during interphase and at the microtubule-kinetochore interface during mitosis. CENP-W depletion causes loss of microtubules and defects in microtubule organization, and both microtubule-stabilizing and -destabilizing agents decrease CENP-W protein stability.","method":"Co-immunoprecipitation; proteasome inhibitor assays; co-localization immunofluorescence; siRNA knockdown; pharmacological microtubule manipulation","journal":"PloS one","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus functional knockdown plus co-localization, single lab","pmids":["26881882"],"is_preprint":false},{"year":2018,"finding":"CENP-W interacts with CUL1 and β-TrCP1 (F-box protein of SCFβ-TrCP1 ubiquitin ligase) through sites overlapping with SKP1 binding; CENP-W incorporation into the SCFβ-TrCP1 complex promotes complex disassembly and β-TrCP1 degradation, thereby decreasing SCFβ-TrCP1 activity. At the G2/M transition, CENP-W knockdown decreases CDC25A protein levels, delaying mitotic entry.","method":"Co-immunoprecipitation; complex disassembly assays; siRNA knockdown with CDC25A protein level and mitotic timing readout; cell cycle synchronization","journal":"FASEB journal","confidence":"High","confidence_rationale":"Tier 2 / Strong — Co-IP plus functional complex disassembly assay plus siRNA phenotype with CDC25A readout, multiple orthogonal methods, clear mechanism","pmids":["29863914"],"is_preprint":false},{"year":2018,"finding":"CENP-W binds both b and f isoforms of β-TrCP1 but with greater affinity for the b isoform; CENP-W (NLS-defective mutant) regulates nuclear-cytoplasmic shuttling of both β-TrCP1 isoforms with preference for isoform b; the Elongin C-binding motif in isoform b contributes to this specificity.","method":"In vivo binding assay (co-transfection/co-IP); fluorescence microscopy of EGFP-β-TrCP1 isoforms with NLS-defective CENP-W mutant","journal":"Genes & genomics","confidence":"Medium","confidence_rationale":"Tier 3 / Moderate — co-IP binding assay plus localization imaging, single lab, limited mechanistic depth","pmids":["30267325"],"is_preprint":false},{"year":2019,"finding":"CENP-W/CUG2-induced stemness-like phenotypes (sphere formation, stemness factor expression) require NPM1 (nucleophosmin): NPM1 suppression by siRNA blocks CUG2-mediated stemness and diminishes TGF-β transcriptional activity and signaling, placing NPM1 upstream of TGF-β in the CUG2 stemness pathway.","method":"siRNA knockdown of NPM1; sphere formation assay; TGF-β reporter assay; Western blotting; epistasis via TGF-β inhibitor and Smad2 siRNA","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — genetic epistasis (NPM1 siRNA plus TGF-β inhibitor), single lab","pmids":["31113615"],"is_preprint":false},{"year":2020,"finding":"In mouse oocytes, CENP-W localizes to the germinal vesicle at GV stage and becomes concentrated on kinetochores during meiotic maturation; siRNA knockdown of CENP-W causes kinetochore-microtubule detachment, defective spindles, chromosome misalignment, metaphase I arrest, failure of first polar body extrusion, and spindle assembly checkpoint activation.","method":"Confocal microscopy (localization); siRNA microinjection in mouse oocytes; immunofluorescence of kinetochore-microtubule attachment; spindle assembly checkpoint assay","journal":"Biochemical and biophysical research communications","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — clean siRNA loss-of-function with specific meiotic phenotypes and localization data, single lab","pmids":["32446395"],"is_preprint":false},{"year":2023,"finding":"Computational modeling (molecular docking, binding free energy calculations, and 250 ns MD simulations with site-directed mutagenesis in silico) identified LEU83 and ARG53 in CENP-W as critical residues for CENP-T/CENP-W heterodimer formation; substitution of these residues with lysine significantly disrupts dimerization.","method":"Molecular docking; binding free energy calculations; molecular dynamics simulation; in silico site-directed mutagenesis","journal":"Journal of cellular biochemistry","confidence":"Low","confidence_rationale":"Tier 4 / Weak — computational prediction only, no experimental validation reported","pmids":["37943107"],"is_preprint":false}],"current_model":"CENPW (also known as CUG2 or C6orf173) encodes an inner kinetochore protein that forms a stable heterodimer with CENP-T; this complex is required for functional kinetochore assembly, kinetochore-microtubule attachment, and accurate chromosome segregation during mitosis and meiosis. CENP-W stability is regulated by the CSN5/JAB1-proteasome axis (which is counteracted by CENP-T complex formation) and by interaction with hnRNP U, while nucleophosmin/B23 controls its proper localization. Beyond kinetochore function, CENP-W modulates cell cycle progression by disassembling the SCFβ-TrCP1 ubiquitin ligase to prevent CDC25A degradation at G2/M, interacts with EZH2 to facilitate Polycomb-mediated gene silencing, and localizes to the nuclear matrix where it associates with RNA—collectively suggesting roles in both mitotic fidelity and transcriptional regulation."},"narrative":{"mechanistic_narrative":"CENPW (also known as CUG2 or C6orf173) encodes an inner kinetochore protein that ensures the fidelity of chromosome segregation in both mitosis and meiosis [PMID:19533040, PMID:32446395]. It forms a centromeric complex with CENP-T and CENP-A, co-localizes with centromeric markers, and is required for accurate chromosome segregation; its depletion causes aberrant cell division, multipolar spindle fragmentation, loss of Hec1 from kinetochores, and acentriolar spindle poles in a manner dependent on microtubules and Eg5 motor activity, indicating it sustains kinetochore-microtubule attachments against motor-generated traction forces [PMID:19533040, PMID:25329824]. In mouse oocytes CENP-W concentrates on kinetochores during meiotic maturation, and its loss produces kinetochore-microtubule detachment, chromosome misalignment, metaphase I arrest, and spindle assembly checkpoint activation [PMID:32446395]. CENP-W abundance is set by proteasomal control: CSN5/JAB1 directly drives ubiquitin-dependent degradation of both CENP-T and CENP-W, while heterodimer formation with CENP-T mutually stabilizes the pair and is required for their kinetochore recruitment [PMID:23926101]. Stability and proper prophase localization are further controlled by nucleophosmin/B23 (NPM1), with which CENP-W physically associates in the nucleolus and nuclear matrix [PMID:22002061], and by hnRNP U, which co-stabilizes CENP-W and co-localizes with it at the microtubule-kinetochore interface [PMID:26881882]. Beyond the kinetochore, CENP-W incorporates into the SCFβ-TrCP1 ubiquitin ligase through SKP1-overlapping sites, promoting complex disassembly and β-TrCP1 degradation, thereby preserving CDC25A levels to permit timely G2/M entry [PMID:29863914], and it associates with EZH2 to enhance its stability and facilitate PRC2-mediated H3K27me3 gene silencing at target promoters [PMID:26111449].","teleology":[{"year":2007,"claim":"Established CENPW as a nuclear protein with oncogenic potential, raising the question of its normal molecular role.","evidence":"EGFP-fusion localization plus soft-agar and nude-mouse xenograft transformation assays in NIH3T3 cells","pmids":["17610844"],"confidence":"Medium","gaps":["No mechanism linking nuclear localization to transformation","Overexpression phenotype only, no loss-of-function"]},{"year":2009,"claim":"Defined CENPW as a centromere component by showing it binds CENP-T and CENP-A and is required for chromosome segregation, anchoring its function at the kinetochore.","evidence":"Yeast two-hybrid, Co-IP, immunofluorescence, and siRNA knockdown with division phenotype","pmids":["19533040"],"confidence":"High","gaps":["Stoichiometry and architecture of the CENP-T/CENP-W complex not resolved","Mechanism of microtubule attachment not addressed"]},{"year":2010,"claim":"Identified Sp1/Sp3 control of the CENPW promoter, explaining how its expression is coupled to proliferative cues.","evidence":"Promoter deletion, EMSA with mutagenesis, and supershift assays under serum stimulation","pmids":["20180024"],"confidence":"Medium","gaps":["Does not connect transcriptional control to cell-cycle timing of CENP-W function"]},{"year":2010,"claim":"Placed CUG2-driven oncogenic signaling through Ras and p38 MAPK by epistasis with selective kinase inhibitors.","evidence":"Pharmacological inhibitor epistasis in stable CUG2-overexpressing cells with viral replication readout","pmids":["20075984"],"confidence":"Medium","gaps":["Direct biochemical link between CENP-W and these kinases not established","Overexpression context only"]},{"year":2011,"claim":"Showed nucleophosmin/B23 controls CENP-W stability and prophase localization, identifying a nucleolar regulator of the protein.","evidence":"Subnuclear fractionation, RNA/DNA association, Co-IP, and B23 siRNA with localization readout","pmids":["22002061"],"confidence":"High","gaps":["Functional consequence of RNA/nuclear-matrix association unresolved","Mechanism of B23-dependent stabilization unknown"]},{"year":2011,"claim":"Demonstrated an in vivo mitotic requirement for cug2 in vertebrate neurogenesis, validating the segregation role in a whole organism.","evidence":"Morpholino knockdown in zebrafish with spindle/chromosome immunofluorescence and apoptosis assays","pmids":["21838932"],"confidence":"Medium","gaps":["Morpholino off-target effects not excluded","Molecular partners in vivo not examined"]},{"year":2013,"claim":"Resolved how CENP-W levels are set, showing CSN5/JAB1 degrades it while CENP-T heterodimerization protects it, coupling complex assembly to kinetochore recruitment.","evidence":"Yeast two-hybrid, Co-IP, ubiquitination/proteasome degradation assays, and kinetochore recruitment imaging","pmids":["23926101"],"confidence":"High","gaps":["Ubiquitin ligase acting downstream of CSN5 not identified","Quantitative balance of degradation vs. stabilization not measured"]},{"year":2014,"claim":"Defined the mechanical role of CENP-W in maintaining kinetochore-microtubule attachments that resist motor-generated forces.","evidence":"RNAi, live imaging, nocodazole/monastrol epistasis, and TPX2 overexpression rescue in HeLa cells","pmids":["25329824"],"confidence":"High","gaps":["Direct role in load-bearing not biochemically measured","Link to centriole splitting mechanism unclear"]},{"year":2015,"claim":"Connected CENP-W to transcriptional repression by showing it stabilizes EZH2 and localizes to PRC2 target promoters.","evidence":"Co-IP, protein stability assays, and ChIP","pmids":["26111449"],"confidence":"Medium","gaps":["Whether EZH2 stabilization is direct not resolved","Genome-wide repression scope not defined"]},{"year":2016,"claim":"Identified hnRNP U as a mutual-stabilization partner linking CENP-W to microtubule organization.","evidence":"Co-IP, proteasome inhibitor assays, co-localization, siRNA, and microtubule drug manipulation","pmids":["26881882"],"confidence":"Medium","gaps":["Reciprocal validation of interaction limited","Mechanism coupling protein stability to microtubule state unknown"]},{"year":2018,"claim":"Uncovered a cell-cycle role distinct from the kinetochore: CENP-W disassembles SCFβ-TrCP1 to preserve CDC25A and enable G2/M entry.","evidence":"Co-IP, complex disassembly assays, and siRNA with CDC25A and mitotic-timing readout","pmids":["29863914"],"confidence":"High","gaps":["How nuclear pool is partitioned between kinetochore and SCF roles unclear","Direct contribution to other β-TrCP1 substrates not assessed"]},{"year":2018,"claim":"Refined β-TrCP1 engagement, showing isoform-selective binding and CENP-W-dependent nuclear-cytoplasmic shuttling of β-TrCP1.","evidence":"Co-transfection binding assays and fluorescence imaging with NLS-defective CENP-W mutant","pmids":["30267325"],"confidence":"Medium","gaps":["Functional significance of isoform preference not tested","Limited mechanistic depth"]},{"year":2019,"claim":"Linked CUG2-induced stemness to an NPM1→TGF-β axis, extending its oncogenic activity to a defined signaling route.","evidence":"NPM1 siRNA, sphere formation, TGF-β reporter, and Smad2 siRNA epistasis","pmids":["31113615"],"confidence":"Medium","gaps":["Direct molecular trigger of TGF-β activity not defined","Overexpression-driven phenotype"]},{"year":2020,"claim":"Established a meiotic requirement, showing CENP-W is needed for kinetochore-microtubule attachment and metaphase I progression in oocytes.","evidence":"Confocal localization and siRNA microinjection in mouse oocytes with attachment and SAC readouts","pmids":["32446395"],"confidence":"Medium","gaps":["Meiosis-specific partners not identified","Single-lab loss-of-function"]},{"year":2023,"claim":"Predicted residues governing CENP-T/CENP-W dimerization, offering a structural hypothesis for complex assembly.","evidence":"Molecular docking, binding free energy, and MD simulations with in silico mutagenesis","pmids":["37943107"],"confidence":"Low","gaps":["Computational only, awaits experimental validation of LEU83/ARG53","No structure of the human complex determined"]},{"year":null,"claim":"How the kinetochore, SCFβ-TrCP1/cell-cycle, and PRC2/transcriptional activities of CENP-W are spatially and temporally coordinated within a single cell remains unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No experimental partitioning of CENP-W pools across its distinct functions","No high-resolution structure of any CENP-W complex"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[6,10]},{"term_id":"GO:0003723","term_label":"RNA binding","supporting_discovery_ids":[4]},{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[1,7]}],"localization":[{"term_id":"GO:0005730","term_label":"nucleolus","supporting_discovery_ids":[4]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[0,4]},{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[1,6]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[1,7,10]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[8]}],"complexes":["CENP-T/CENP-W heterodimer","SCFβ-TrCP1 ubiquitin ligase","kinetochore"],"partners":["CENPT","CENPA","NPM1","CSN5/JAB1","HNRNPU","EZH2","BTRC","CUL1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q5EE01","full_name":"Centromere protein W","aliases":["Cancer-up-regulated gene 2 protein"],"length_aa":88,"mass_kda":10.1,"function":"Component of the CENPA-NAC (nucleosome-associated) complex, a complex that plays a central role in assembly of kinetochore proteins, mitotic progression and chromosome segregation (By similarity). The CENPA-NAC complex recruits the CENPA-CAD (nucleosome distal) complex and may be involved in incorporation of newly synthesized CENPA into centromeres (By similarity). Part of a nucleosome-associated complex that binds specifically to histone H3-containing nucleosomes at the centromere, as opposed to nucleosomes containing CENPA. Component of the heterotetrameric CENP-T-W-S-X complex that binds and supercoils DNA, and plays an important role in kinetochore assembly. CENPW has a fundamental role in kinetochore assembly and function. It is one of the inner kinetochore proteins, with most further proteins binding downstream. Required for normal chromosome organization and normal progress through mitosis","subcellular_location":"Nucleus; Chromosome, centromere; Chromosome, centromere, kinetochore; Nucleus matrix; Nucleus, nucleolus","url":"https://www.uniprot.org/uniprotkb/Q5EE01/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CENPW","classification":"Common Essential","n_dependent_lines":1196,"n_total_lines":1208,"dependency_fraction":0.9900662251655629},"opencell":{"profiled":true,"resolved_as":"","ensg_id":"ENSG00000203760","cell_line_id":"CID000927","localizations":[{"compartment":"nuclear_punctae","grade":3}],"interactors":[],"url":"https://opencell.sf.czbiohub.org/target/CID000927","total_profiled":1310},"omim":[{"mim_id":"611510","title":"CENTROMERIC PROTEIN T; CENPT","url":"https://www.omim.org/entry/611510"},{"mim_id":"611264","title":"CENTROMERIC PROTEIN W; CENPW","url":"https://www.omim.org/entry/611264"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nucleoplasm","reliability":"Supported"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in all","driving_tissues":[{"tissue":"bone marrow","ntpm":47.7}],"url":"https://www.proteinatlas.org/search/CENPW"},"hgnc":{"alias_symbol":["CUG2"],"prev_symbol":["C6orf173"]},"alphafold":{"accession":"Q5EE01","domains":[{"cath_id":"1.10.20.10","chopping":"36-85","consensus_level":"high","plddt":96.285,"start":36,"end":85}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5EE01","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q5EE01-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q5EE01-F1-predicted_aligned_error_v6.png","plddt_mean":89.69},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CENPW","jax_strain_url":"https://www.jax.org/strain/search?query=CENPW"},"sequence":{"accession":"Q5EE01","fasta_url":"https://rest.uniprot.org/uniprotkb/Q5EE01.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q5EE01/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q5EE01"}},"corpus_meta":[{"pmid":"17610844","id":"PMC_17610844","title":"Molecular cloning and functional analysis of a novel oncogene, cancer-upregulated gene 2 (CUG2).","date":"2007","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/17610844","citation_count":41,"is_preprint":false},{"pmid":"30226605","id":"PMC_30226605","title":"STAT1‑HDAC4 signaling induces epithelial‑mesenchymal transition and sphere formation of cancer cells overexpressing the oncogene, CUG2.","date":"2018","source":"Oncology reports","url":"https://pubmed.ncbi.nlm.nih.gov/30226605","citation_count":36,"is_preprint":false},{"pmid":"19533040","id":"PMC_19533040","title":"Cancer-upregulated gene 2 (CUG2), a new component of centromere complex, is required for kinetochore function.","date":"2009","source":"Molecules and cells","url":"https://pubmed.ncbi.nlm.nih.gov/19533040","citation_count":32,"is_preprint":false},{"pmid":"28776259","id":"PMC_28776259","title":"Increased EGFR expression induced by a novel oncogene, CUG2, confers resistance to doxorubicin through Stat1-HDAC4 signaling.","date":"2017","source":"Cellular oncology (Dordrecht, Netherlands)","url":"https://pubmed.ncbi.nlm.nih.gov/28776259","citation_count":28,"is_preprint":false},{"pmid":"20075984","id":"PMC_20075984","title":"CUG2, a novel oncogene confers reoviral replication through Ras and p38 signaling pathway.","date":"2010","source":"Cancer gene therapy","url":"https://pubmed.ncbi.nlm.nih.gov/20075984","citation_count":25,"is_preprint":false},{"pmid":"22002061","id":"PMC_22002061","title":"New centromeric component CENP-W is an RNA-associated nuclear matrix protein that interacts with nucleophosmin/B23 protein.","date":"2011","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/22002061","citation_count":25,"is_preprint":false},{"pmid":"30771899","id":"PMC_30771899","title":"Cancer upregulated gene (CUG)2 elevates YAP1 expression, leading to enhancement of epithelial-mesenchymal transition in human lung cancer cells.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/30771899","citation_count":22,"is_preprint":false},{"pmid":"32635817","id":"PMC_32635817","title":"Histone-fold centromere protein W (CENP-W) is associated with the biological behavior of hepatocellular carcinoma cells.","date":"2020","source":"Bioengineered","url":"https://pubmed.ncbi.nlm.nih.gov/32635817","citation_count":21,"is_preprint":false},{"pmid":"33973496","id":"PMC_33973496","title":"Knockdown of CENPW Inhibits Hepatocellular Carcinoma Progression by Inactivating E2F Signaling.","date":"2021","source":"Technology in cancer research & treatment","url":"https://pubmed.ncbi.nlm.nih.gov/33973496","citation_count":17,"is_preprint":false},{"pmid":"31113615","id":"PMC_31113615","title":"Cancer upregulated gene 2 (CUG2), a novel oncogene, promotes stemness-like properties via the NPM1-TGF-β signaling axis.","date":"2019","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/31113615","citation_count":16,"is_preprint":false},{"pmid":"26881882","id":"PMC_26881882","title":"Centromere Protein (CENP)-W Interacts with Heterogeneous Nuclear Ribonucleoprotein (hnRNP) U and May Contribute to Kinetochore-Microtubule Attachment in Mitotic Cells.","date":"2016","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/26881882","citation_count":16,"is_preprint":false},{"pmid":"23926101","id":"PMC_23926101","title":"CSN5/JAB1 interacts with the centromeric components CENP-T and CENP-W and regulates their proteasome-mediated degradation.","date":"2013","source":"The Journal of biological chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/23926101","citation_count":15,"is_preprint":false},{"pmid":"29863914","id":"PMC_29863914","title":"CENP-W inhibits CDC25A degradation by destabilizing the SCFβ-TrCP-1 complex at G2/M.","date":"2018","source":"FASEB journal : official publication of the Federation of American Societies for Experimental Biology","url":"https://pubmed.ncbi.nlm.nih.gov/29863914","citation_count":11,"is_preprint":false},{"pmid":"24452380","id":"PMC_24452380","title":"Suppression of autophagic genes sensitizes CUG2-overexpressing A549 human lung cancer cells to oncolytic vesicular stomatitis virus-induced apoptosis.","date":"2014","source":"International journal of oncology","url":"https://pubmed.ncbi.nlm.nih.gov/24452380","citation_count":10,"is_preprint":false},{"pmid":"30287155","id":"PMC_30287155","title":"N-Benzyl-N-methyl-dodecan-1-amine, a novel compound from garlic, exerts anti-cancer effects on human A549 lung cancer cells overexpressing cancer upregulated gene (CUG)2.","date":"2018","source":"European journal of pharmacology","url":"https://pubmed.ncbi.nlm.nih.gov/30287155","citation_count":9,"is_preprint":false},{"pmid":"20180024","id":"PMC_20180024","title":"Sp1 and Sp3 mediate basal and serum-induced expression of human CENP-W.","date":"2010","source":"Molecular biology reports","url":"https://pubmed.ncbi.nlm.nih.gov/20180024","citation_count":9,"is_preprint":false},{"pmid":"25329824","id":"PMC_25329824","title":"CENP-W plays a role in maintaining bipolar spindle structure.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25329824","citation_count":9,"is_preprint":false},{"pmid":"33116878","id":"PMC_33116878","title":"Overexpression of Cancer Upregulated Gene 2 (CUG2) Decreases Spry2 Through c-Cbl, Leading to Activation of EGFR and β-Catenin Signaling.","date":"2020","source":"Cancer management and research","url":"https://pubmed.ncbi.nlm.nih.gov/33116878","citation_count":8,"is_preprint":false},{"pmid":"26111449","id":"PMC_26111449","title":"A new kinetochore component CENP-W interacts with the polycomb-group protein EZH2 to promote gene silencing.","date":"2015","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/26111449","citation_count":8,"is_preprint":false},{"pmid":"38213721","id":"PMC_38213721","title":"CENPW knockdown inhibits progression of bladder cancer through inducing cell cycle arrest and apoptosis.","date":"2024","source":"Journal of Cancer","url":"https://pubmed.ncbi.nlm.nih.gov/38213721","citation_count":6,"is_preprint":false},{"pmid":"20648695","id":"PMC_20648695","title":"Cancer-upregulated gene 2 (CUG2) overexpression induces apoptosis in SKOV-3 cells.","date":"2010","source":"Cell biochemistry and function","url":"https://pubmed.ncbi.nlm.nih.gov/20648695","citation_count":6,"is_preprint":false},{"pmid":"21838932","id":"PMC_21838932","title":"Cug2 is essential for normal mitotic control and CNS development in zebrafish.","date":"2011","source":"BMC developmental biology","url":"https://pubmed.ncbi.nlm.nih.gov/21838932","citation_count":5,"is_preprint":false},{"pmid":"32446395","id":"PMC_32446395","title":"CENP-W regulates kinetochore-microtubule attachment and meiotic progression of mouse oocytes.","date":"2020","source":"Biochemical and biophysical research communications","url":"https://pubmed.ncbi.nlm.nih.gov/32446395","citation_count":5,"is_preprint":false},{"pmid":"37727713","id":"PMC_37727713","title":"Rare ROS1-CENPW gene in pancreatic acinar cell carcinoma and the effect of crizotinib plus AG chemotherapy: A case report.","date":"2023","source":"World journal of clinical cases","url":"https://pubmed.ncbi.nlm.nih.gov/37727713","citation_count":4,"is_preprint":false},{"pmid":"30267325","id":"PMC_30267325","title":"Two major alternative splice variants of beta-TrCP1 interact with CENP-W with different binding preferences.","date":"2018","source":"Genes & genomics","url":"https://pubmed.ncbi.nlm.nih.gov/30267325","citation_count":2,"is_preprint":false},{"pmid":"37943107","id":"PMC_37943107","title":"Bioinformatics insights into CENP-T and CENP-W protein-protein interaction disruptive amino acid substitution in the CENP-T-W complex.","date":"2023","source":"Journal of cellular biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/37943107","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":14578,"output_tokens":3868,"usd":0.050877,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":11624,"output_tokens":4242,"usd":0.082085,"stage2_stop_reason":"end_turn"},"total_usd":0.132962,"stage1_batch_id":"msgbatch_01ACAxJQabDsht39JXeWG22d","stage2_batch_id":"msgbatch_01QEU22A9U5sbPTgsMzTmvSf","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"CENPW (CUG2) protein localizes predominantly to the nucleus when expressed as EGFP fusion, and overexpression in NIH3T3 mouse fibroblasts induces cancer-specific phenotypes in vitro and tumor formation in nude mice, establishing proto-oncogenic activity.\",\n      \"method\": \"EGFP fusion protein localization imaging; soft-agar and nude mouse xenograft assays\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct localization experiment and functional KO/OE assay, single lab, single study\",\n      \"pmids\": [\"17610844\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2009,\n      \"finding\": \"CENPW (CUG2) physically interacts with CENP-T (a component of the CENP-A nucleosome complex) and CENP-A in a centromeric complex, co-localizes with centromeric markers, and is required for proper chromosome segregation during mitosis; its depletion induces aberrant cell division.\",\n      \"method\": \"Yeast two-hybrid screening; co-immunoprecipitation; immunofluorescent staining; siRNA knockdown with cell viability and division phenotype readout\",\n      \"journal\": \"Molecules and cells\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — yeast two-hybrid plus Co-IP plus immunofluorescence plus functional knockdown, replicated across multiple subsequent studies\",\n      \"pmids\": [\"19533040\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"The CENPW promoter contains a GC-rich Sp1-binding site (-46 to -36) that is critical for basal and serum-induced expression; Sp1 and Sp3 transcription factors specifically bind this site and mediate transactivation of CENPW.\",\n      \"method\": \"Promoter deletion analysis; competitive EMSA with mutated oligos; supershift assays with Sp1/Sp3 antibodies; serum stimulation experiments\",\n      \"journal\": \"Molecular biology reports\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — EMSA with mutagenesis and supershift, single lab, two orthogonal methods\",\n      \"pmids\": [\"20180024\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2010,\n      \"finding\": \"CUG2 expression activates MAPK (ERK, JNK, p38), Src kinase, and Ras signaling; inhibition of Ras or p38 MAPK (but not ERK, JNK, or Src) blocks reoviral replication in CUG2-expressing cells, establishing Ras and p38 as necessary downstream effectors of CUG2-mediated permissiveness to reovirus.\",\n      \"method\": \"Pharmacological inhibitors of specific kinases; stable CUG2-overexpressing NIH3T3 cells; viral replication assays\",\n      \"journal\": \"Cancer gene therapy\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — epistasis via selective inhibitors with clear pathway placement, single lab\",\n      \"pmids\": [\"20075984\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"CENPW (CENP-W) localizes to the nucleolus and nuclear matrix, associates with both RNA and DNA by fractionation, and physically interacts with the nucleolar phosphoprotein nucleophosmin (B23/NPM1); depletion of B23 by siRNA decreases CENP-W protein stability and causes its severe mislocalization during prophase.\",\n      \"method\": \"Subnuclear fractionation; RNA/DNA association assays; biochemical affinity binding (co-IP); siRNA knockdown; immunofluorescence microscopy\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (fractionation, affinity binding, siRNA with localization phenotype) in single rigorous study\",\n      \"pmids\": [\"22002061\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2011,\n      \"finding\": \"In zebrafish, morpholino-mediated knockdown of cug2 causes mitotic arrest with abnormal spindle formation and chromosome misalignment in the neural plate, followed by CNS-wide apoptosis, establishing an in vivo requirement for Cug2 in normal mitosis during neurogenesis.\",\n      \"method\": \"Morpholino knockdown in zebrafish embryos; spindle/chromosome immunofluorescence; apoptosis assays\",\n      \"journal\": \"BMC developmental biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean in vivo loss-of-function with defined mitotic phenotype, single lab\",\n      \"pmids\": [\"21838932\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"CSN5/JAB1 directly interacts with both CENP-T and CENP-W (identified by yeast two-hybrid and confirmed by Co-IP), promotes their ubiquitin- and proteasome-dependent degradation, and formation of the CENP-T·CENP-W complex stabilizes both proteins by blocking CSN5-mediated degradation; dysregulation of CSN5 causes severe defects in CENP-T·CENP-W recruitment to the kinetochore during prophase.\",\n      \"method\": \"Yeast two-hybrid; co-immunoprecipitation; ubiquitination/proteasome degradation assays; immunofluorescence of kinetochore recruitment\",\n      \"journal\": \"The Journal of biological chemistry\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — yeast two-hybrid plus Co-IP plus functional degradation assays plus localization phenotype, multiple orthogonal methods in single study\",\n      \"pmids\": [\"23926101\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CENP-W depletion in HeLa cells causes bipolar spindle fragmentation into multipolar spindles, depletion of Hec1 at kinetochores, abnormal centriole splitting, and generation of acentriolar spindle poles; spindle pole fragmentation requires microtubules (absent in nocodazole) and Eg5 motor activity (reduced by monastrol), implicating CENP-W in maintaining kinetochore-microtubule attachment that resists motor-generated traction forces.\",\n      \"method\": \"RNAi depletion; live-cell fluorescence imaging (H2B and tubulin); immunofluorescence of centrioles and centrosomal markers; pharmacological manipulation (nocodazole, monastrol); TPX2 overexpression rescue\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — multiple orthogonal methods (RNAi, live imaging, pharmacological epistasis, genetic rescue) in single rigorous study\",\n      \"pmids\": [\"25329824\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CENP-W physically associates with EZH2 (catalytic subunit of PRC2), enhances EZH2 protein stability, and is recruited to the promoters of EZH2 target genes (by chromatin immunoprecipitation) to facilitate EZH2-mediated transcriptional repression (H3K27me3-associated gene silencing).\",\n      \"method\": \"Co-immunoprecipitation; protein stability assays; chromatin immunoprecipitation (ChIP)\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus ChIP, single lab, two orthogonal methods\",\n      \"pmids\": [\"26111449\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"CENP-W physically interacts with hnRNP U; the interaction mutually stabilizes both proteins by inhibiting proteasome-mediated degradation. They co-localize in the nuclear matrix during interphase and at the microtubule-kinetochore interface during mitosis. CENP-W depletion causes loss of microtubules and defects in microtubule organization, and both microtubule-stabilizing and -destabilizing agents decrease CENP-W protein stability.\",\n      \"method\": \"Co-immunoprecipitation; proteasome inhibitor assays; co-localization immunofluorescence; siRNA knockdown; pharmacological microtubule manipulation\",\n      \"journal\": \"PloS one\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus functional knockdown plus co-localization, single lab\",\n      \"pmids\": [\"26881882\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CENP-W interacts with CUL1 and β-TrCP1 (F-box protein of SCFβ-TrCP1 ubiquitin ligase) through sites overlapping with SKP1 binding; CENP-W incorporation into the SCFβ-TrCP1 complex promotes complex disassembly and β-TrCP1 degradation, thereby decreasing SCFβ-TrCP1 activity. At the G2/M transition, CENP-W knockdown decreases CDC25A protein levels, delaying mitotic entry.\",\n      \"method\": \"Co-immunoprecipitation; complex disassembly assays; siRNA knockdown with CDC25A protein level and mitotic timing readout; cell cycle synchronization\",\n      \"journal\": \"FASEB journal\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — Co-IP plus functional complex disassembly assay plus siRNA phenotype with CDC25A readout, multiple orthogonal methods, clear mechanism\",\n      \"pmids\": [\"29863914\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CENP-W binds both b and f isoforms of β-TrCP1 but with greater affinity for the b isoform; CENP-W (NLS-defective mutant) regulates nuclear-cytoplasmic shuttling of both β-TrCP1 isoforms with preference for isoform b; the Elongin C-binding motif in isoform b contributes to this specificity.\",\n      \"method\": \"In vivo binding assay (co-transfection/co-IP); fluorescence microscopy of EGFP-β-TrCP1 isoforms with NLS-defective CENP-W mutant\",\n      \"journal\": \"Genes & genomics\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 / Moderate — co-IP binding assay plus localization imaging, single lab, limited mechanistic depth\",\n      \"pmids\": [\"30267325\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CENP-W/CUG2-induced stemness-like phenotypes (sphere formation, stemness factor expression) require NPM1 (nucleophosmin): NPM1 suppression by siRNA blocks CUG2-mediated stemness and diminishes TGF-β transcriptional activity and signaling, placing NPM1 upstream of TGF-β in the CUG2 stemness pathway.\",\n      \"method\": \"siRNA knockdown of NPM1; sphere formation assay; TGF-β reporter assay; Western blotting; epistasis via TGF-β inhibitor and Smad2 siRNA\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — genetic epistasis (NPM1 siRNA plus TGF-β inhibitor), single lab\",\n      \"pmids\": [\"31113615\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"In mouse oocytes, CENP-W localizes to the germinal vesicle at GV stage and becomes concentrated on kinetochores during meiotic maturation; siRNA knockdown of CENP-W causes kinetochore-microtubule detachment, defective spindles, chromosome misalignment, metaphase I arrest, failure of first polar body extrusion, and spindle assembly checkpoint activation.\",\n      \"method\": \"Confocal microscopy (localization); siRNA microinjection in mouse oocytes; immunofluorescence of kinetochore-microtubule attachment; spindle assembly checkpoint assay\",\n      \"journal\": \"Biochemical and biophysical research communications\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean siRNA loss-of-function with specific meiotic phenotypes and localization data, single lab\",\n      \"pmids\": [\"32446395\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"Computational modeling (molecular docking, binding free energy calculations, and 250 ns MD simulations with site-directed mutagenesis in silico) identified LEU83 and ARG53 in CENP-W as critical residues for CENP-T/CENP-W heterodimer formation; substitution of these residues with lysine significantly disrupts dimerization.\",\n      \"method\": \"Molecular docking; binding free energy calculations; molecular dynamics simulation; in silico site-directed mutagenesis\",\n      \"journal\": \"Journal of cellular biochemistry\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 4 / Weak — computational prediction only, no experimental validation reported\",\n      \"pmids\": [\"37943107\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CENPW (also known as CUG2 or C6orf173) encodes an inner kinetochore protein that forms a stable heterodimer with CENP-T; this complex is required for functional kinetochore assembly, kinetochore-microtubule attachment, and accurate chromosome segregation during mitosis and meiosis. CENP-W stability is regulated by the CSN5/JAB1-proteasome axis (which is counteracted by CENP-T complex formation) and by interaction with hnRNP U, while nucleophosmin/B23 controls its proper localization. Beyond kinetochore function, CENP-W modulates cell cycle progression by disassembling the SCFβ-TrCP1 ubiquitin ligase to prevent CDC25A degradation at G2/M, interacts with EZH2 to facilitate Polycomb-mediated gene silencing, and localizes to the nuclear matrix where it associates with RNA—collectively suggesting roles in both mitotic fidelity and transcriptional regulation.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CENPW (also known as CUG2 or C6orf173) encodes an inner kinetochore protein that ensures the fidelity of chromosome segregation in both mitosis and meiosis [#1, #13]. It forms a centromeric complex with CENP-T and CENP-A, co-localizes with centromeric markers, and is required for accurate chromosome segregation; its depletion causes aberrant cell division, multipolar spindle fragmentation, loss of Hec1 from kinetochores, and acentriolar spindle poles in a manner dependent on microtubules and Eg5 motor activity, indicating it sustains kinetochore-microtubule attachments against motor-generated traction forces [#1, #7]. In mouse oocytes CENP-W concentrates on kinetochores during meiotic maturation, and its loss produces kinetochore-microtubule detachment, chromosome misalignment, metaphase I arrest, and spindle assembly checkpoint activation [#13]. CENP-W abundance is set by proteasomal control: CSN5/JAB1 directly drives ubiquitin-dependent degradation of both CENP-T and CENP-W, while heterodimer formation with CENP-T mutually stabilizes the pair and is required for their kinetochore recruitment [#6]. Stability and proper prophase localization are further controlled by nucleophosmin/B23 (NPM1), with which CENP-W physically associates in the nucleolus and nuclear matrix [#4], and by hnRNP U, which co-stabilizes CENP-W and co-localizes with it at the microtubule-kinetochore interface [#9]. Beyond the kinetochore, CENP-W incorporates into the SCFβ-TrCP1 ubiquitin ligase through SKP1-overlapping sites, promoting complex disassembly and β-TrCP1 degradation, thereby preserving CDC25A levels to permit timely G2/M entry [#10], and it associates with EZH2 to enhance its stability and facilitate PRC2-mediated H3K27me3 gene silencing at target promoters [#8].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"Established CENPW as a nuclear protein with oncogenic potential, raising the question of its normal molecular role.\",\n      \"evidence\": \"EGFP-fusion localization plus soft-agar and nude-mouse xenograft transformation assays in NIH3T3 cells\",\n      \"pmids\": [\"17610844\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No mechanism linking nuclear localization to transformation\", \"Overexpression phenotype only, no loss-of-function\"]\n    },\n    {\n      \"year\": 2009,\n      \"claim\": \"Defined CENPW as a centromere component by showing it binds CENP-T and CENP-A and is required for chromosome segregation, anchoring its function at the kinetochore.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, immunofluorescence, and siRNA knockdown with division phenotype\",\n      \"pmids\": [\"19533040\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Stoichiometry and architecture of the CENP-T/CENP-W complex not resolved\", \"Mechanism of microtubule attachment not addressed\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Identified Sp1/Sp3 control of the CENPW promoter, explaining how its expression is coupled to proliferative cues.\",\n      \"evidence\": \"Promoter deletion, EMSA with mutagenesis, and supershift assays under serum stimulation\",\n      \"pmids\": [\"20180024\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Does not connect transcriptional control to cell-cycle timing of CENP-W function\"]\n    },\n    {\n      \"year\": 2010,\n      \"claim\": \"Placed CUG2-driven oncogenic signaling through Ras and p38 MAPK by epistasis with selective kinase inhibitors.\",\n      \"evidence\": \"Pharmacological inhibitor epistasis in stable CUG2-overexpressing cells with viral replication readout\",\n      \"pmids\": [\"20075984\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct biochemical link between CENP-W and these kinases not established\", \"Overexpression context only\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Showed nucleophosmin/B23 controls CENP-W stability and prophase localization, identifying a nucleolar regulator of the protein.\",\n      \"evidence\": \"Subnuclear fractionation, RNA/DNA association, Co-IP, and B23 siRNA with localization readout\",\n      \"pmids\": [\"22002061\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Functional consequence of RNA/nuclear-matrix association unresolved\", \"Mechanism of B23-dependent stabilization unknown\"]\n    },\n    {\n      \"year\": 2011,\n      \"claim\": \"Demonstrated an in vivo mitotic requirement for cug2 in vertebrate neurogenesis, validating the segregation role in a whole organism.\",\n      \"evidence\": \"Morpholino knockdown in zebrafish with spindle/chromosome immunofluorescence and apoptosis assays\",\n      \"pmids\": [\"21838932\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Morpholino off-target effects not excluded\", \"Molecular partners in vivo not examined\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Resolved how CENP-W levels are set, showing CSN5/JAB1 degrades it while CENP-T heterodimerization protects it, coupling complex assembly to kinetochore recruitment.\",\n      \"evidence\": \"Yeast two-hybrid, Co-IP, ubiquitination/proteasome degradation assays, and kinetochore recruitment imaging\",\n      \"pmids\": [\"23926101\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Ubiquitin ligase acting downstream of CSN5 not identified\", \"Quantitative balance of degradation vs. stabilization not measured\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Defined the mechanical role of CENP-W in maintaining kinetochore-microtubule attachments that resist motor-generated forces.\",\n      \"evidence\": \"RNAi, live imaging, nocodazole/monastrol epistasis, and TPX2 overexpression rescue in HeLa cells\",\n      \"pmids\": [\"25329824\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct role in load-bearing not biochemically measured\", \"Link to centriole splitting mechanism unclear\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"Connected CENP-W to transcriptional repression by showing it stabilizes EZH2 and localizes to PRC2 target promoters.\",\n      \"evidence\": \"Co-IP, protein stability assays, and ChIP\",\n      \"pmids\": [\"26111449\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether EZH2 stabilization is direct not resolved\", \"Genome-wide repression scope not defined\"]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Identified hnRNP U as a mutual-stabilization partner linking CENP-W to microtubule organization.\",\n      \"evidence\": \"Co-IP, proteasome inhibitor assays, co-localization, siRNA, and microtubule drug manipulation\",\n      \"pmids\": [\"26881882\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Reciprocal validation of interaction limited\", \"Mechanism coupling protein stability to microtubule state unknown\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Uncovered a cell-cycle role distinct from the kinetochore: CENP-W disassembles SCFβ-TrCP1 to preserve CDC25A and enable G2/M entry.\",\n      \"evidence\": \"Co-IP, complex disassembly assays, and siRNA with CDC25A and mitotic-timing readout\",\n      \"pmids\": [\"29863914\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"How nuclear pool is partitioned between kinetochore and SCF roles unclear\", \"Direct contribution to other β-TrCP1 substrates not assessed\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Refined β-TrCP1 engagement, showing isoform-selective binding and CENP-W-dependent nuclear-cytoplasmic shuttling of β-TrCP1.\",\n      \"evidence\": \"Co-transfection binding assays and fluorescence imaging with NLS-defective CENP-W mutant\",\n      \"pmids\": [\"30267325\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Functional significance of isoform preference not tested\", \"Limited mechanistic depth\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Linked CUG2-induced stemness to an NPM1→TGF-β axis, extending its oncogenic activity to a defined signaling route.\",\n      \"evidence\": \"NPM1 siRNA, sphere formation, TGF-β reporter, and Smad2 siRNA epistasis\",\n      \"pmids\": [\"31113615\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct molecular trigger of TGF-β activity not defined\", \"Overexpression-driven phenotype\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Established a meiotic requirement, showing CENP-W is needed for kinetochore-microtubule attachment and metaphase I progression in oocytes.\",\n      \"evidence\": \"Confocal localization and siRNA microinjection in mouse oocytes with attachment and SAC readouts\",\n      \"pmids\": [\"32446395\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Meiosis-specific partners not identified\", \"Single-lab loss-of-function\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Predicted residues governing CENP-T/CENP-W dimerization, offering a structural hypothesis for complex assembly.\",\n      \"evidence\": \"Molecular docking, binding free energy, and MD simulations with in silico mutagenesis\",\n      \"pmids\": [\"37943107\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Computational only, awaits experimental validation of LEU83/ARG53\", \"No structure of the human complex determined\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the kinetochore, SCFβ-TrCP1/cell-cycle, and PRC2/transcriptional activities of CENP-W are spatially and temporally coordinated within a single cell remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No experimental partitioning of CENP-W pools across its distinct functions\", \"No high-resolution structure of any CENP-W complex\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [6, 10]},\n      {\"term_id\": \"GO:0003723\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [1, 7]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005730\", \"supporting_discovery_ids\": [4]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [0, 4]},\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [1, 6]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [1, 7, 10]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [8]}\n    ],\n    \"complexes\": [\n      \"CENP-T/CENP-W heterodimer\",\n      \"SCFβ-TrCP1 ubiquitin ligase\",\n      \"kinetochore\"\n    ],\n    \"partners\": [\n      \"CENPT\",\n      \"CENPA\",\n      \"NPM1\",\n      \"CSN5/JAB1\",\n      \"HNRNPU\",\n      \"EZH2\",\n      \"BTRC\",\n      \"CUL1\"\n    ],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":6,"faith_total":6,"faith_pct":100.0}}