{"gene":"CENPK","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2009,"finding":"CENP-K and CENP-H form a stable subcomplex with approximately 1:1 stoichiometry, even under high-salt conditions, as demonstrated by tandem affinity purification (TAP) from HEK 293 cells expressing TAP-CENP-K. Bioinformatic analysis indicated both proteins are enriched in coiled-coil regions, and functional mapping showed their N- and C-terminal regions mediate the interaction, suggesting they form heterodimeric coiled-coils within the inner kinetochore.","method":"Tandem affinity purification (TAP) from HEK 293 cells; bioinformatic coiled-coil prediction; deletion mapping of interacting regions","journal":"Science in China. Series C, Life sciences","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — reciprocal co-purification by TAP with stoichiometric evidence, supported by bioinformatic structural prediction and domain mapping, but single lab and no in vitro reconstitution or structural validation","pmids":["19381461"],"is_preprint":false},{"year":2017,"finding":"Overexpression of CENP-K in HCC cells stimulated tyrosine phosphorylation of AKT and MDM2 proteins while inhibiting tyrosine phosphorylation of TP53, placing CENP-K upstream of the AKT/MDM2/TP53 signaling axis in promoting cell proliferation.","method":"Western blot for phosphorylation; overexpression and knockdown in SMMC7721, Focus, MHCC-LM3, and QGY7703 cell lines; proliferation and migration assays","journal":"Oncotarget","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, single method (western blot for phosphorylation), no direct binding or enzymatic assay establishing mechanism","pmids":["29088763"],"is_preprint":false},{"year":2019,"finding":"CENPK knockdown suppressed proliferation, migration, invasion, and EMT in hepatocellular carcinoma cells; these inhibitory effects were partially rescued by restoring YAP1 expression, placing CENPK upstream of YAP1 in regulating HCC malignant progression.","method":"shRNA knockdown; Cell Counting Kit-8, colony formation, wound healing, transwell invasion assays; Western blot for EMT markers and YAP1; genetic rescue experiment with YAP1 re-expression","journal":"OncoTargets and therapy","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, genetic rescue establishes pathway order but no direct molecular interaction between CENPK and YAP1 was demonstrated","pmids":["30774374"],"is_preprint":false},{"year":2021,"finding":"CENPK knockdown in gastric cancer cells decreased expression of PI3K, phospho-AKT (Ser437), and phospho-GSK3β (Ser9) while increasing PTEN expression, indicating CENPK promotes cell growth and survival through the PTEN-PI3K-AKT signaling pathway.","method":"shRNA knockdown; Western blot for pathway components; proliferation assays in vitro; xenograft tumor growth in vivo; KEGG pathway analysis","journal":"Journal of cellular and molecular medicine","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, pathway association by western blot without direct molecular interaction or epistasis rescue","pmids":["34382342"],"is_preprint":false},{"year":2022,"finding":"CENPK physically interacts with XRCC5 (Ku80) in gastric cancer cells, as identified by Co-immunoprecipitation followed by LC-MS proteomics; functional rescue assays confirmed that XRCC5 mediates the pro-proliferative and pro-migratory effects of CENPK in gastric cancer cells.","method":"Co-immunoprecipitation; LC-MS proteomics; functional rescue assay; proliferation, migration, invasion assays; flow cytometry for apoptosis and cell cycle; xenograft model","journal":"Gastric cancer","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — Co-IP plus LC-MS identification plus functional rescue in a single lab, providing multiple orthogonal methods to establish the CENPK–XRCC5 interaction and its functional relevance","pmids":["35715658"],"is_preprint":false},{"year":2022,"finding":"ZC3H13-mediated m6A modification of CENPK mRNA promotes CENPK expression in cervical cancer. CENPK protein directly binds SOX6 and disrupts CENPK–β-catenin interactions, leading to increased β-catenin nuclear translocation, p53 ubiquitination, and activation of Wnt/β-catenin signaling with suppression of the p53 pathway, thereby enhancing stemness, chemoresistance, and EMT.","method":"Methylated RNA immunoprecipitation (MeRIP) for m6A detection; co-immunoprecipitation for CENPK–SOX6 and CENPK–β-catenin interactions; chromatin immunoprecipitation; luciferase reporter assay; cycloheximide chase assay; cell fractionation; Western blot; tumorsphere formation; clonogenic and xenograft assays","journal":"Military Medical Research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (MeRIP, Co-IP, ChIP, luciferase, fractionation) in a single lab establishing both the m6A regulatory mechanism and the downstream CENPK–SOX6–β-catenin pathway","pmids":["35418160"],"is_preprint":false},{"year":2022,"finding":"CENPK knockdown in colorectal cancer cells was associated with upregulation of CUL4A (Cullin 4A), and overexpression of CUL4A partially rescued the anti-proliferative effects of CENPK silencing, suggesting CENPK acts upstream of CUL4A-mediated regulation in CRC.","method":"shRNA lentiviral knockdown; qPCR; Western blot; MTT assay; flow cytometry; xenograft fluorescence imaging in vivo; CUL4A overexpression rescue experiment","journal":"World journal of gastroenterology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, rescue experiment suggests pathway order but no direct molecular interaction demonstrated between CENPK and CUL4A","pmids":["36312839"],"is_preprint":false},{"year":2024,"finding":"A splice variant of CENPK lacking exon 8 (CENPK-delta8) specifically binds FLNA (filamin A) and FLOT1 (flotillin-1), interactions not observed with wild-type CENPK, linking CENPK-delta8 to cytoskeleton organization and cell migration and conferring abiraterone resistance in metastatic castration-resistant prostate cancer.","method":"Protein binding assays identifying FLNA and FLOT1 as CENPK-delta8-specific interactors; patient-derived xenograft (PDX) models; 3D organoids from responders and non-responders; in vitro migration and proliferation assays","journal":"Cells","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — specific binding partners identified for the splice variant vs wild-type, validated in PDX and organoid models as well as in vitro, single lab but multiple model systems","pmids":["39404386"],"is_preprint":false},{"year":2024,"finding":"E2F1 transcription factor directly regulates CENPK transcription in ovarian cancer; CENPK silencing suppresses the mTOR pathway; CENPK promotes sensitivity to the mTOR inhibitor rapamycin; CENPK interacts with GOLPH3 to mediate mTOR signaling activation.","method":"Transcription factor binding/reporter assays establishing E2F1 as CENPK regulator; CENPK siRNA knockdown with mTOR pathway readouts by Western blot; rapamycin sensitivity assays; co-immunoprecipitation/interaction assays with GOLPH3","journal":"Molecular and cellular endocrinology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — single lab, mechanistic details not fully described in abstract; E2F1 regulation and GOLPH3 interaction inferred from functional and reporter assays without full structural or biochemical validation","pmids":["38670220"],"is_preprint":false},{"year":2026,"finding":"CTCF directly binds to the CENPK promoter and positively regulates its transcription (confirmed by ChIP-qPCR and luciferase reporter assay). The CTCF–CENPK axis activates JAK1/STAT3 signaling (increased JAK1 and STAT3 phosphorylation). Zeylenone directly binds CTCF (thermal shift assay; molecular docking), inhibiting CTCF-mediated CENPK transcription and thereby suppressing JAK1/STAT3 pathway activity and tamoxifen resistance in breast cancer.","method":"ChIP-qPCR; luciferase reporter assay; thermal shift assay for Zey–CTCF binding; molecular docking; Western blot for JAK1/STAT3 phosphorylation; CENPK and JAK1 knockdown rescue experiments; in vitro and in vivo tumor models","journal":"Drug development research","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal methods (ChIP, luciferase, thermal shift, rescue experiments) in a single lab establishing CTCF as a transcriptional regulator of CENPK and the downstream JAK1/STAT3 pathway connection","pmids":["41797275"],"is_preprint":false}],"current_model":"CENPK (CENP-K) is a constitutive centromere-associated network (CCAN) protein that forms a stable heterodimeric coiled-coil subcomplex with CENP-H at the inner kinetochore; in cancer contexts, its transcription is driven by E2F1 and CTCF, its mRNA is stabilized by ZC3H13-mediated m6A modification, and its protein promotes cell proliferation, migration, and survival by interacting with partners including XRCC5, SOX6/β-catenin, GOLPH3/mTOR, and—in a specific exon-8-deleted splice variant—FLNA and FLOT1, collectively activating AKT, Wnt/β-catenin, JAK/STAT3, and mTOR signaling pathways."},"narrative":{"mechanistic_narrative":"CENPK is an inner kinetochore protein that forms a stable, salt-resistant heterodimeric coiled-coil subcomplex with CENP-H at approximately 1:1 stoichiometry within the constitutive centromere-associated network [PMID:19381461]. Beyond this structural role, the available corpus characterizes CENPK predominantly as a transcriptionally controlled driver of malignant cell behavior across multiple cancers. Its transcription is directly activated by E2F1 [PMID:38670220] and CTCF [PMID:41797275], and its mRNA is stabilized by ZC3H13-mediated m6A modification [PMID:35418160]. At the protein level, CENPK engages distinct partners to activate parallel pro-tumorigenic signaling axes: it binds XRCC5 (Ku80) to promote proliferation and migration [PMID:35715658], binds SOX6 and competitively disrupts CENPK–β-catenin interactions to drive β-catenin nuclear translocation, p53 ubiquitination, and Wnt/β-catenin activation [PMID:35418160], and interacts with GOLPH3 to activate mTOR signaling and confer rapamycin sensitivity [PMID:38670220]. An exon-8-deleted splice variant (CENPK-delta8) acquires specific binding to FLNA and FLOT1 not seen with wild-type protein, linking it to cytoskeletal organization, migration, and abiraterone resistance in prostate cancer [PMID:39404386]. The CTCF–CENPK axis additionally activates JAK1/STAT3 signaling [PMID:41797275]. Beyond the kinetochore heterodimer with CENP-H [PMID:19381461], the mechanistic basis by which CENPK transduces these many signaling outputs has not been resolved into a unified biochemical model in the available corpus.","teleology":[{"year":2009,"claim":"Established that CENP-K is a stable structural component of the inner kinetochore by defining its direct partnership with CENP-H, addressing how these CCAN proteins assemble.","evidence":"Tandem affinity purification from HEK293 cells with coiled-coil prediction and deletion mapping of interacting regions","pmids":["19381461"],"confidence":"Medium","gaps":["No in vitro reconstitution or structural validation of the heterodimer","Stoichiometry and architecture within the full CCAN not resolved","Single lab"]},{"year":2017,"claim":"First linked CENPK overexpression to oncogenic signaling, placing it upstream of the AKT/MDM2/TP53 axis in hepatocellular carcinoma proliferation.","evidence":"Overexpression/knockdown with western blot for phosphorylation and proliferation/migration assays in HCC lines","pmids":["29088763"],"confidence":"Low","gaps":["Single method (phospho-western), no direct binding or enzymatic mechanism","How CENPK affects phosphorylation is unknown"]},{"year":2019,"claim":"Positioned CENPK upstream of YAP1 in driving HCC malignant progression and EMT via a genetic rescue, establishing pathway order without molecular contact.","evidence":"shRNA knockdown with functional assays and YAP1 re-expression rescue in HCC cells","pmids":["30774374"],"confidence":"Low","gaps":["No direct CENPK–YAP1 interaction demonstrated","Mechanism connecting CENPK to YAP1 unknown"]},{"year":2021,"claim":"Connected CENPK to the PTEN-PI3K-AKT pathway in gastric cancer, refining the signaling context for its pro-survival role.","evidence":"shRNA knockdown with pathway westerns, in vitro proliferation and xenograft assays","pmids":["34382342"],"confidence":"Low","gaps":["Pathway association by western blot only, no direct interaction or epistasis rescue","Causal node unidentified"]},{"year":2022,"claim":"Identified XRCC5 (Ku80) as a direct physical partner mediating CENPK's pro-proliferative and pro-migratory effects in gastric cancer.","evidence":"Co-IP plus LC-MS proteomics with functional rescue and xenograft validation","pmids":["35715658"],"confidence":"Medium","gaps":["Interaction interface and stoichiometry not mapped","Mechanistic consequence of binding not biochemically defined"]},{"year":2022,"claim":"Defined both an upstream regulatory mechanism (ZC3H13/m6A mRNA stabilization) and a downstream effector pathway, showing CENPK binds SOX6 and disrupts CENPK–β-catenin interactions to drive Wnt/β-catenin activation and p53 suppression in cervical cancer.","evidence":"MeRIP, Co-IP, ChIP, luciferase, cycloheximide chase and fractionation with tumorsphere and xenograft assays","pmids":["35418160"],"confidence":"Medium","gaps":["Structural basis of the competitive SOX6/β-catenin binding not resolved","Generality beyond cervical cancer untested"]},{"year":2022,"claim":"Placed CENPK upstream of CUL4A regulation in colorectal cancer proliferation via a rescue experiment.","evidence":"shRNA knockdown with qPCR, western, MTT, flow cytometry, xenograft imaging, and CUL4A overexpression rescue","pmids":["36312839"],"confidence":"Low","gaps":["No direct CENPK–CUL4A interaction shown","Mechanism of CUL4A regulation unknown"]},{"year":2024,"claim":"Revealed a splice-variant-specific interactome, with CENPK-delta8 acquiring FLNA and FLOT1 binding absent in wild-type protein, linking the isoform to cytoskeletal organization and abiraterone resistance.","evidence":"Variant-specific protein binding assays validated in PDX models, 3D organoids, and in vitro migration assays","pmids":["39404386"],"confidence":"Medium","gaps":["Structural basis for exon-8-dependent partner specificity unknown","Single lab"]},{"year":2024,"claim":"Connected E2F1-driven CENPK transcription to mTOR pathway activation via a GOLPH3 interaction, explaining rapamycin sensitivity in ovarian cancer.","evidence":"TF reporter assays, siRNA knockdown with mTOR readouts, rapamycin sensitivity and Co-IP with GOLPH3","pmids":["38670220"],"confidence":"Low","gaps":["GOLPH3 interaction not structurally validated","Mechanism linking CENPK–GOLPH3 to mTOR not fully described"]},{"year":2026,"claim":"Established CTCF as a direct transcriptional activator of CENPK driving JAK1/STAT3 signaling and tamoxifen resistance, and identified Zeylenone as a pharmacological inhibitor of this axis.","evidence":"ChIP-qPCR, luciferase reporter, thermal shift assay, molecular docking, and CENPK/JAK1 knockdown rescue in vitro and in vivo","pmids":["41797275"],"confidence":"Medium","gaps":["How CENPK protein mechanistically activates JAK1/STAT3 not defined","Single lab"]},{"year":null,"claim":"It remains unresolved how a single inner-kinetochore protein mechanistically transduces such diverse and partner-specific signaling outputs (Wnt, mTOR, JAK/STAT, PI3K/AKT) and whether these activities are separable from its CCAN structural role.","evidence":"","pmids":[],"confidence":"Low","gaps":["No structural model of CENPK signaling complexes","Relationship between kinetochore function and oncogenic signaling unexplored","Most downstream pathway links rest on single-lab western/rescue data"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0005198","term_label":"structural molecule activity","supporting_discovery_ids":[0]},{"term_id":"GO:0060089","term_label":"molecular transducer activity","supporting_discovery_ids":[4,5,8]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0]},{"term_id":"R-HSA-162582","term_label":"Signal Transduction","supporting_discovery_ids":[5,8,9]},{"term_id":"R-HSA-1643685","term_label":"Disease","supporting_discovery_ids":[4,5,7,9]}],"complexes":["CENP-H/CENP-K subcomplex","CCAN (constitutive centromere-associated network)"],"partners":["CENPH","XRCC5","SOX6","CTNNB1","GOLPH3","FLNA","FLOT1"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q9BS16","full_name":"Centromere protein K","aliases":["Interphase centromere complex protein 37","Protein AF-5alpha","p33"],"length_aa":269,"mass_kda":31.7,"function":"Component of the CENPA-CAD (nucleosome distal) complex, a complex recruited to centromeres which is involved in assembly of kinetochore proteins, mitotic progression and chromosome segregation. May be involved in incorporation of newly synthesized CENPA into centromeres via its interaction with the CENPA-NAC complex. Acts in coordination with KNL1 to recruit the NDC80 complex to the outer kinetochore","subcellular_location":"Nucleus; Chromosome, centromere; Chromosome, centromere, kinetochore","url":"https://www.uniprot.org/uniprotkb/Q9BS16/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CENPK","classification":"Common Essential","n_dependent_lines":1200,"n_total_lines":1208,"dependency_fraction":0.9933774834437086},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CENPK","total_profiled":1310},"omim":[{"mim_id":"611503","title":"CENTROMERIC PROTEIN L; CENPL","url":"https://www.omim.org/entry/611503"},{"mim_id":"611502","title":"CENTROMERIC PROTEIN K; CENPK","url":"https://www.omim.org/entry/611502"},{"mim_id":"610152","title":"CENTROMERIC PROTEIN M; CENPM","url":"https://www.omim.org/entry/610152"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"","locations":[],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":23.5},{"tissue":"lymphoid tissue","ntpm":20.7}],"url":"https://www.proteinatlas.org/search/CENPK"},"hgnc":{"alias_symbol":["FKSG14","SOLT","CENP-K"],"prev_symbol":[]},"alphafold":{"accession":"Q9BS16","domains":[{"cath_id":"-","chopping":"209-269","consensus_level":"medium","plddt":87.5302,"start":209,"end":269},{"cath_id":"1.20.5","chopping":"86-154","consensus_level":"medium","plddt":93.8648,"start":86,"end":154},{"cath_id":"1.20.5","chopping":"155-188","consensus_level":"medium","plddt":91.5012,"start":155,"end":188}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BS16","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BS16-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9BS16-F1-predicted_aligned_error_v6.png","plddt_mean":81.0},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CENPK","jax_strain_url":"https://www.jax.org/strain/search?query=CENPK"},"sequence":{"accession":"Q9BS16","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9BS16.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9BS16/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9BS16"}},"corpus_meta":[{"pmid":"35418160","id":"PMC_35418160","title":"N6-methyladenosine modification of CENPK mRNA by ZC3H13 promotes cervical cancer stemness and chemoresistance.","date":"2022","source":"Military Medical Research","url":"https://pubmed.ncbi.nlm.nih.gov/35418160","citation_count":92,"is_preprint":false},{"pmid":"16201081","id":"PMC_16201081","title":"Hepatocyte growth factor as well as vascular endothelial growth factor gene induction effectively promotes liver regeneration after hepatectomy in Solt-Farber rats.","date":"2005","source":"Hepato-gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/16201081","citation_count":38,"is_preprint":false},{"pmid":"30774374","id":"PMC_30774374","title":"Downregulation of CENPK suppresses hepatocellular carcinoma malignant progression through regulating YAP1.","date":"2019","source":"OncoTargets and therapy","url":"https://pubmed.ncbi.nlm.nih.gov/30774374","citation_count":31,"is_preprint":false},{"pmid":"3552207","id":"PMC_3552207","title":"Coordinate polypeptide expression during hepatocarcinogenesis in male F-344 rats: comparison of the Solt-Farber and Reddy models.","date":"1987","source":"Cancer research","url":"https://pubmed.ncbi.nlm.nih.gov/3552207","citation_count":28,"is_preprint":false},{"pmid":"33658048","id":"PMC_33658048","title":"LINC00958 promotes the proliferation of TSCC via miR-211-5p/CENPK axis and activating the JAK/STAT3 signaling pathway.","date":"2021","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/33658048","citation_count":27,"is_preprint":false},{"pmid":"29088763","id":"PMC_29088763","title":"Overexpression of centromere protein K (CENP-K) gene in hepatocellular carcinoma promote cell proliferation by activating AKT/TP53 signal pathway.","date":"2017","source":"Oncotarget","url":"https://pubmed.ncbi.nlm.nih.gov/29088763","citation_count":24,"is_preprint":false},{"pmid":"10996314","id":"PMC_10996314","title":"Characterization of Solt, a novel SoxLZ/Sox6 binding protein expressed in adult mouse testis.","date":"2000","source":"FEBS letters","url":"https://pubmed.ncbi.nlm.nih.gov/10996314","citation_count":20,"is_preprint":false},{"pmid":"34382342","id":"PMC_34382342","title":"Knockdown of CENPK inhibits cell growth and facilitates apoptosis via PTEN-PI3K-AKT signalling pathway in gastric cancer.","date":"2021","source":"Journal of cellular and molecular medicine","url":"https://pubmed.ncbi.nlm.nih.gov/34382342","citation_count":15,"is_preprint":false},{"pmid":"30662630","id":"PMC_30662630","title":"Diwu Yanggan capsule inhibits the occurrence and development of liver cancer in the Solt-Farber rat model by regulating the Ras/Raf/Mek/Erk signaling pathway.","date":"2018","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/30662630","citation_count":10,"is_preprint":false},{"pmid":"33904381","id":"PMC_33904381","title":"Overexpression of centromere protein K (CENPK) gene in Differentiated Thyroid Carcinoma promote cell Proliferation and Migration.","date":"2021","source":"Bioengineered","url":"https://pubmed.ncbi.nlm.nih.gov/33904381","citation_count":9,"is_preprint":false},{"pmid":"19381461","id":"PMC_19381461","title":"CENP-K and CENP-H may form coiled-coils in the kinetochores.","date":"2009","source":"Science in China. Series C, Life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/19381461","citation_count":8,"is_preprint":false},{"pmid":"35715658","id":"PMC_35715658","title":"Centromeric protein K (CENPK) promotes gastric cancer proliferation and migration via interacting with XRCC5.","date":"2022","source":"Gastric cancer : official journal of the International Gastric Cancer Association and the Japanese Gastric Cancer Association","url":"https://pubmed.ncbi.nlm.nih.gov/35715658","citation_count":7,"is_preprint":false},{"pmid":"36312839","id":"PMC_36312839","title":"Lentivirus-mediated short hairpin RNA interference of CENPK inhibits growth of colorectal cancer cells with overexpression of Cullin 4A.","date":"2022","source":"World journal of gastroenterology","url":"https://pubmed.ncbi.nlm.nih.gov/36312839","citation_count":6,"is_preprint":false},{"pmid":"35035680","id":"PMC_35035680","title":"Pan-cancer investigation of CENPK gene: clinical significance and oncogenic immunology.","date":"2021","source":"American journal of translational research","url":"https://pubmed.ncbi.nlm.nih.gov/35035680","citation_count":5,"is_preprint":false},{"pmid":"34044826","id":"PMC_34044826","title":"LINC01158 works as an oncogene in glioma via sponging miR-6734-3p to boost CENPK expression.","date":"2021","source":"Cancer cell international","url":"https://pubmed.ncbi.nlm.nih.gov/34044826","citation_count":4,"is_preprint":false},{"pmid":"38670220","id":"PMC_38670220","title":"CENPK orchestrates ovarian cancer progression via GOLPH3-Mediated activation of mTOR signaling.","date":"2024","source":"Molecular and cellular endocrinology","url":"https://pubmed.ncbi.nlm.nih.gov/38670220","citation_count":3,"is_preprint":false},{"pmid":"39404386","id":"PMC_39404386","title":"The Role of CENPK Splice Variant in Abiraterone Response in Metastatic Castration-Resistant Prostate Cancer.","date":"2024","source":"Cells","url":"https://pubmed.ncbi.nlm.nih.gov/39404386","citation_count":1,"is_preprint":false},{"pmid":"41797275","id":"PMC_41797275","title":"Zeylenone Attenuates Tamoxifen Resistance by Directly Binding to CTCF and Inhibiting the CTCF-CENPK-JAK1/STAT3 Signaling Axis in Breast Cancer.","date":"2026","source":"Drug development research","url":"https://pubmed.ncbi.nlm.nih.gov/41797275","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11766,"output_tokens":2966,"usd":0.039894,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":10323,"output_tokens":3369,"usd":0.06792,"stage2_stop_reason":"end_turn"},"total_usd":0.107814,"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\": 2009,\n      \"finding\": \"CENP-K and CENP-H form a stable subcomplex with approximately 1:1 stoichiometry, even under high-salt conditions, as demonstrated by tandem affinity purification (TAP) from HEK 293 cells expressing TAP-CENP-K. Bioinformatic analysis indicated both proteins are enriched in coiled-coil regions, and functional mapping showed their N- and C-terminal regions mediate the interaction, suggesting they form heterodimeric coiled-coils within the inner kinetochore.\",\n      \"method\": \"Tandem affinity purification (TAP) from HEK 293 cells; bioinformatic coiled-coil prediction; deletion mapping of interacting regions\",\n      \"journal\": \"Science in China. Series C, Life sciences\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — reciprocal co-purification by TAP with stoichiometric evidence, supported by bioinformatic structural prediction and domain mapping, but single lab and no in vitro reconstitution or structural validation\",\n      \"pmids\": [\"19381461\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"Overexpression of CENP-K in HCC cells stimulated tyrosine phosphorylation of AKT and MDM2 proteins while inhibiting tyrosine phosphorylation of TP53, placing CENP-K upstream of the AKT/MDM2/TP53 signaling axis in promoting cell proliferation.\",\n      \"method\": \"Western blot for phosphorylation; overexpression and knockdown in SMMC7721, Focus, MHCC-LM3, and QGY7703 cell lines; proliferation and migration assays\",\n      \"journal\": \"Oncotarget\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, single method (western blot for phosphorylation), no direct binding or enzymatic assay establishing mechanism\",\n      \"pmids\": [\"29088763\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2019,\n      \"finding\": \"CENPK knockdown suppressed proliferation, migration, invasion, and EMT in hepatocellular carcinoma cells; these inhibitory effects were partially rescued by restoring YAP1 expression, placing CENPK upstream of YAP1 in regulating HCC malignant progression.\",\n      \"method\": \"shRNA knockdown; Cell Counting Kit-8, colony formation, wound healing, transwell invasion assays; Western blot for EMT markers and YAP1; genetic rescue experiment with YAP1 re-expression\",\n      \"journal\": \"OncoTargets and therapy\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, genetic rescue establishes pathway order but no direct molecular interaction between CENPK and YAP1 was demonstrated\",\n      \"pmids\": [\"30774374\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"CENPK knockdown in gastric cancer cells decreased expression of PI3K, phospho-AKT (Ser437), and phospho-GSK3β (Ser9) while increasing PTEN expression, indicating CENPK promotes cell growth and survival through the PTEN-PI3K-AKT signaling pathway.\",\n      \"method\": \"shRNA knockdown; Western blot for pathway components; proliferation assays in vitro; xenograft tumor growth in vivo; KEGG pathway analysis\",\n      \"journal\": \"Journal of cellular and molecular medicine\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, pathway association by western blot without direct molecular interaction or epistasis rescue\",\n      \"pmids\": [\"34382342\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CENPK physically interacts with XRCC5 (Ku80) in gastric cancer cells, as identified by Co-immunoprecipitation followed by LC-MS proteomics; functional rescue assays confirmed that XRCC5 mediates the pro-proliferative and pro-migratory effects of CENPK in gastric cancer cells.\",\n      \"method\": \"Co-immunoprecipitation; LC-MS proteomics; functional rescue assay; proliferation, migration, invasion assays; flow cytometry for apoptosis and cell cycle; xenograft model\",\n      \"journal\": \"Gastric cancer\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — Co-IP plus LC-MS identification plus functional rescue in a single lab, providing multiple orthogonal methods to establish the CENPK–XRCC5 interaction and its functional relevance\",\n      \"pmids\": [\"35715658\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"ZC3H13-mediated m6A modification of CENPK mRNA promotes CENPK expression in cervical cancer. CENPK protein directly binds SOX6 and disrupts CENPK–β-catenin interactions, leading to increased β-catenin nuclear translocation, p53 ubiquitination, and activation of Wnt/β-catenin signaling with suppression of the p53 pathway, thereby enhancing stemness, chemoresistance, and EMT.\",\n      \"method\": \"Methylated RNA immunoprecipitation (MeRIP) for m6A detection; co-immunoprecipitation for CENPK–SOX6 and CENPK–β-catenin interactions; chromatin immunoprecipitation; luciferase reporter assay; cycloheximide chase assay; cell fractionation; Western blot; tumorsphere formation; clonogenic and xenograft assays\",\n      \"journal\": \"Military Medical Research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (MeRIP, Co-IP, ChIP, luciferase, fractionation) in a single lab establishing both the m6A regulatory mechanism and the downstream CENPK–SOX6–β-catenin pathway\",\n      \"pmids\": [\"35418160\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"CENPK knockdown in colorectal cancer cells was associated with upregulation of CUL4A (Cullin 4A), and overexpression of CUL4A partially rescued the anti-proliferative effects of CENPK silencing, suggesting CENPK acts upstream of CUL4A-mediated regulation in CRC.\",\n      \"method\": \"shRNA lentiviral knockdown; qPCR; Western blot; MTT assay; flow cytometry; xenograft fluorescence imaging in vivo; CUL4A overexpression rescue experiment\",\n      \"journal\": \"World journal of gastroenterology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, rescue experiment suggests pathway order but no direct molecular interaction demonstrated between CENPK and CUL4A\",\n      \"pmids\": [\"36312839\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"A splice variant of CENPK lacking exon 8 (CENPK-delta8) specifically binds FLNA (filamin A) and FLOT1 (flotillin-1), interactions not observed with wild-type CENPK, linking CENPK-delta8 to cytoskeleton organization and cell migration and conferring abiraterone resistance in metastatic castration-resistant prostate cancer.\",\n      \"method\": \"Protein binding assays identifying FLNA and FLOT1 as CENPK-delta8-specific interactors; patient-derived xenograft (PDX) models; 3D organoids from responders and non-responders; in vitro migration and proliferation assays\",\n      \"journal\": \"Cells\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — specific binding partners identified for the splice variant vs wild-type, validated in PDX and organoid models as well as in vitro, single lab but multiple model systems\",\n      \"pmids\": [\"39404386\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"E2F1 transcription factor directly regulates CENPK transcription in ovarian cancer; CENPK silencing suppresses the mTOR pathway; CENPK promotes sensitivity to the mTOR inhibitor rapamycin; CENPK interacts with GOLPH3 to mediate mTOR signaling activation.\",\n      \"method\": \"Transcription factor binding/reporter assays establishing E2F1 as CENPK regulator; CENPK siRNA knockdown with mTOR pathway readouts by Western blot; rapamycin sensitivity assays; co-immunoprecipitation/interaction assays with GOLPH3\",\n      \"journal\": \"Molecular and cellular endocrinology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — single lab, mechanistic details not fully described in abstract; E2F1 regulation and GOLPH3 interaction inferred from functional and reporter assays without full structural or biochemical validation\",\n      \"pmids\": [\"38670220\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2026,\n      \"finding\": \"CTCF directly binds to the CENPK promoter and positively regulates its transcription (confirmed by ChIP-qPCR and luciferase reporter assay). The CTCF–CENPK axis activates JAK1/STAT3 signaling (increased JAK1 and STAT3 phosphorylation). Zeylenone directly binds CTCF (thermal shift assay; molecular docking), inhibiting CTCF-mediated CENPK transcription and thereby suppressing JAK1/STAT3 pathway activity and tamoxifen resistance in breast cancer.\",\n      \"method\": \"ChIP-qPCR; luciferase reporter assay; thermal shift assay for Zey–CTCF binding; molecular docking; Western blot for JAK1/STAT3 phosphorylation; CENPK and JAK1 knockdown rescue experiments; in vitro and in vivo tumor models\",\n      \"journal\": \"Drug development research\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal methods (ChIP, luciferase, thermal shift, rescue experiments) in a single lab establishing CTCF as a transcriptional regulator of CENPK and the downstream JAK1/STAT3 pathway connection\",\n      \"pmids\": [\"41797275\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CENPK (CENP-K) is a constitutive centromere-associated network (CCAN) protein that forms a stable heterodimeric coiled-coil subcomplex with CENP-H at the inner kinetochore; in cancer contexts, its transcription is driven by E2F1 and CTCF, its mRNA is stabilized by ZC3H13-mediated m6A modification, and its protein promotes cell proliferation, migration, and survival by interacting with partners including XRCC5, SOX6/β-catenin, GOLPH3/mTOR, and—in a specific exon-8-deleted splice variant—FLNA and FLOT1, collectively activating AKT, Wnt/β-catenin, JAK/STAT3, and mTOR signaling pathways.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CENPK is an inner kinetochore protein that forms a stable, salt-resistant heterodimeric coiled-coil subcomplex with CENP-H at approximately 1:1 stoichiometry within the constitutive centromere-associated network [#0]. Beyond this structural role, the available corpus characterizes CENPK predominantly as a transcriptionally controlled driver of malignant cell behavior across multiple cancers. Its transcription is directly activated by E2F1 [#8] and CTCF [#9], and its mRNA is stabilized by ZC3H13-mediated m6A modification [#5]. At the protein level, CENPK engages distinct partners to activate parallel pro-tumorigenic signaling axes: it binds XRCC5 (Ku80) to promote proliferation and migration [#4], binds SOX6 and competitively disrupts CENPK–\\u03b2-catenin interactions to drive \\u03b2-catenin nuclear translocation, p53 ubiquitination, and Wnt/\\u03b2-catenin activation [#5], and interacts with GOLPH3 to activate mTOR signaling and confer rapamycin sensitivity [#8]. An exon-8-deleted splice variant (CENPK-delta8) acquires specific binding to FLNA and FLOT1 not seen with wild-type protein, linking it to cytoskeletal organization, migration, and abiraterone resistance in prostate cancer [#7]. The CTCF–CENPK axis additionally activates JAK1/STAT3 signaling [#9]. Beyond the kinetochore heterodimer with CENP-H [#0], the mechanistic basis by which CENPK transduces these many signaling outputs has not been resolved into a unified biochemical model in the available corpus.\",\n  \"teleology\": [\n    {\n      \"year\": 2009,\n      \"claim\": \"Established that CENP-K is a stable structural component of the inner kinetochore by defining its direct partnership with CENP-H, addressing how these CCAN proteins assemble.\",\n      \"evidence\": \"Tandem affinity purification from HEK293 cells with coiled-coil prediction and deletion mapping of interacting regions\",\n      \"pmids\": [\"19381461\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No in vitro reconstitution or structural validation of the heterodimer\", \"Stoichiometry and architecture within the full CCAN not resolved\", \"Single lab\"]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"First linked CENPK overexpression to oncogenic signaling, placing it upstream of the AKT/MDM2/TP53 axis in hepatocellular carcinoma proliferation.\",\n      \"evidence\": \"Overexpression/knockdown with western blot for phosphorylation and proliferation/migration assays in HCC lines\",\n      \"pmids\": [\"29088763\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Single method (phospho-western), no direct binding or enzymatic mechanism\", \"How CENPK affects phosphorylation is unknown\"]\n    },\n    {\n      \"year\": 2019,\n      \"claim\": \"Positioned CENPK upstream of YAP1 in driving HCC malignant progression and EMT via a genetic rescue, establishing pathway order without molecular contact.\",\n      \"evidence\": \"shRNA knockdown with functional assays and YAP1 re-expression rescue in HCC cells\",\n      \"pmids\": [\"30774374\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct CENPK–YAP1 interaction demonstrated\", \"Mechanism connecting CENPK to YAP1 unknown\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Connected CENPK to the PTEN-PI3K-AKT pathway in gastric cancer, refining the signaling context for its pro-survival role.\",\n      \"evidence\": \"shRNA knockdown with pathway westerns, in vitro proliferation and xenograft assays\",\n      \"pmids\": [\"34382342\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"Pathway association by western blot only, no direct interaction or epistasis rescue\", \"Causal node unidentified\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified XRCC5 (Ku80) as a direct physical partner mediating CENPK's pro-proliferative and pro-migratory effects in gastric cancer.\",\n      \"evidence\": \"Co-IP plus LC-MS proteomics with functional rescue and xenograft validation\",\n      \"pmids\": [\"35715658\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interaction interface and stoichiometry not mapped\", \"Mechanistic consequence of binding not biochemically defined\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Defined both an upstream regulatory mechanism (ZC3H13/m6A mRNA stabilization) and a downstream effector pathway, showing CENPK binds SOX6 and disrupts CENPK–\\u03b2-catenin interactions to drive Wnt/\\u03b2-catenin activation and p53 suppression in cervical cancer.\",\n      \"evidence\": \"MeRIP, Co-IP, ChIP, luciferase, cycloheximide chase and fractionation with tumorsphere and xenograft assays\",\n      \"pmids\": [\"35418160\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis of the competitive SOX6/\\u03b2-catenin binding not resolved\", \"Generality beyond cervical cancer untested\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Placed CENPK upstream of CUL4A regulation in colorectal cancer proliferation via a rescue experiment.\",\n      \"evidence\": \"shRNA knockdown with qPCR, western, MTT, flow cytometry, xenograft imaging, and CUL4A overexpression rescue\",\n      \"pmids\": [\"36312839\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct CENPK–CUL4A interaction shown\", \"Mechanism of CUL4A regulation unknown\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Revealed a splice-variant-specific interactome, with CENPK-delta8 acquiring FLNA and FLOT1 binding absent in wild-type protein, linking the isoform to cytoskeletal organization and abiraterone resistance.\",\n      \"evidence\": \"Variant-specific protein binding assays validated in PDX models, 3D organoids, and in vitro migration assays\",\n      \"pmids\": [\"39404386\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Structural basis for exon-8-dependent partner specificity unknown\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Connected E2F1-driven CENPK transcription to mTOR pathway activation via a GOLPH3 interaction, explaining rapamycin sensitivity in ovarian cancer.\",\n      \"evidence\": \"TF reporter assays, siRNA knockdown with mTOR readouts, rapamycin sensitivity and Co-IP with GOLPH3\",\n      \"pmids\": [\"38670220\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"GOLPH3 interaction not structurally validated\", \"Mechanism linking CENPK–GOLPH3 to mTOR not fully described\"]\n    },\n    {\n      \"year\": 2026,\n      \"claim\": \"Established CTCF as a direct transcriptional activator of CENPK driving JAK1/STAT3 signaling and tamoxifen resistance, and identified Zeylenone as a pharmacological inhibitor of this axis.\",\n      \"evidence\": \"ChIP-qPCR, luciferase reporter, thermal shift assay, molecular docking, and CENPK/JAK1 knockdown rescue in vitro and in vivo\",\n      \"pmids\": [\"41797275\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"How CENPK protein mechanistically activates JAK1/STAT3 not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"It remains unresolved how a single inner-kinetochore protein mechanistically transduces such diverse and partner-specific signaling outputs (Wnt, mTOR, JAK/STAT, PI3K/AKT) and whether these activities are separable from its CCAN structural role.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No structural model of CENPK signaling complexes\", \"Relationship between kinetochore function and oncogenic signaling unexplored\", \"Most downstream pathway links rest on single-lab western/rescue data\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0005198\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"GO:0060089\", \"supporting_discovery_ids\": [4, 5, 8]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [5, 8, 9]},\n      {\"term_id\": \"R-HSA-1643685\", \"supporting_discovery_ids\": [4, 5, 7, 9]}\n    ],\n    \"complexes\": [\"CENP-H/CENP-K subcomplex\", \"CCAN (constitutive centromere-associated network)\"],\n    \"partners\": [\"CENPH\", \"XRCC5\", \"SOX6\", \"CTNNB1\", \"GOLPH3\", \"FLNA\", \"FLOT1\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":3,"faith_total":5,"faith_pct":60.0}}