{"gene":"CENPL","run_date":"2026-06-09T22:57:18","timeline":{"discoveries":[{"year":2006,"finding":"CENP-L (CENPL) is a component of the CENP-A nucleosome-associated complex (NAC) at human centromeres. CENP-A nucleosomes directly recruit the NAC, and CENP-L is identified as one of seven CENP-A-nucleosome distal (CAD) components assembling on the NAC. Disruption of the NAC causes chromosome alignment and segregation errors that preclude cell survival.","method":"Affinity purification / mass spectrometry (AP-MS) of CENP-A nucleosome complexes; RNAi depletion with mitotic phenotype readout","journal":"Nature Cell Biology","confidence":"High","confidence_rationale":"Tier 2 / Strong — reciprocal affinity purification with MS identification, functional depletion experiments, replicated by subsequent studies","pmids":["16622419"],"is_preprint":false},{"year":2012,"finding":"CENPL is essential for the stability of kinetochore microtubules, for a homogeneous poleward microtubule flux rate, and for the kinetochore pushing force that accelerates centrosome separation in prometaphase. CENP-L depletion causes massive chromosome alignment and segregation defects due to syntelic and merotelic kinetochore-microtubule attachments.","method":"siRNA depletion of CENP-L in human cells; live-cell imaging and quantitative measurement of centrosome separation kinetics, microtubule flux, and kinetochore-microtubule attachment geometry","journal":"Journal of Cell Science","confidence":"High","confidence_rationale":"Tier 2 / Moderate — clean KO/KD with defined cellular phenotypes (microtubule stability, flux, attachment fidelity), multiple orthogonal readouts in a single study","pmids":["22399803"],"is_preprint":false},{"year":2012,"finding":"CENP-P/O/R/Q/U sub-complex binding to the CCAN is largely mediated through interactions with CENP-L (as the CENP-N binding partner) and CENP-K. CENP-L thus acts as a hub connecting the CENP-N module to the CENP-PORQU sub-complex.","method":"Fluorescent three-hybrid (F3H) assay, FRET in living mammalian cells, and co-immunoprecipitation to map protein-protein interactions","journal":"PloS One","confidence":"Medium","confidence_rationale":"Tier 2 / Moderate — multiple orthogonal interaction assays (F3H, FRET, Co-IP) in a single study, single lab","pmids":["23028590"],"is_preprint":false},{"year":2018,"finding":"CENP-L co-migrates with CENP-K in the nucleoplasm outside centromeres in living human interphase cells, demonstrating that CENP-L and CENP-K form part of a soluble CENP-C/H/I/K/M/T/W/N/L complex in the nucleoplasm independently of centromere binding.","method":"Fluorescence cross-correlation spectroscopy (FCCS) in living human interphase cells","journal":"PloS One","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — direct in-cell biophysical measurement (FCCS) establishing co-migration, single lab, single method","pmids":["29509805"],"is_preprint":false},{"year":2022,"finding":"Phosphorylation of CENP-L and CENP-N controls CENP-LN complex formation and centromere localization in a cell cycle-dependent manner. Mimicking constitutive phosphorylation of either CENP-L or CENP-N, or simultaneously preventing phosphorylation of both, prevents CENP-LN localization and disrupts chromosome segregation. Cycles of phosphorylation and dephosphorylation are critical for CENP-LN complex recruitment and CCAN reorganization across the cell cycle.","method":"Phosphomimetic and phospho-null mutagenesis of CENP-L and CENP-N; live-cell imaging; chromosome segregation assays","journal":"Molecular Biology of the Cell","confidence":"High","confidence_rationale":"Tier 1-2 / Moderate — direct mutagenesis of phosphosites with functional readouts (localization and chromosome segregation), multiple mutant conditions tested, single lab","pmids":["35830614"],"is_preprint":false},{"year":2013,"finding":"The yeast CENP-L ortholog Iml3 adopts an elongated conformation with intramolecular interactions (crystal structure resolved). Its binding partner Chl4 (CENP-N ortholog) C-terminal domain forms a dimer and is responsible for Iml3 binding. The Chl4-Iml3 heterodimer exhibits low-affinity nonspecific DNA-binding activity, potentially relevant to kinetochore assembly.","method":"X-ray crystallography of Iml3; pull-down assays mapping Chl4-Iml3 interaction domains; in vitro DNA-binding assays","journal":"Acta Crystallographica Section D","confidence":"Medium","confidence_rationale":"Tier 1 / Weak — crystal structure and in vitro biochemistry for the yeast ortholog, not directly demonstrated for human CENPL","pmids":["24311582"],"is_preprint":false},{"year":2024,"finding":"CENPL is a transcriptional target of E2F8 and mediates E2F8's role in homologous recombination (HR) repair of DNA double-strand breaks and chemoresistance. CENPL overexpression in E2F8-knockdown cells partially rescued HR repair and chemotherapy sensitivity; CENPL knockdown alone impaired HR repair and sensitized cells to DNA-damaging drugs.","method":"siRNA knockdown and overexpression of CENPL and E2F8; HR repair assays; chemosensitivity assays in breast cancer cells; transcriptional target validation","journal":"Cellular Signalling","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — rescue epistasis experiment placing CENPL downstream of E2F8 in HR repair, single lab, multiple functional assays","pmids":["38522807"],"is_preprint":false},{"year":2023,"finding":"CENPL activates the MEK1/2-ERK1/2 signaling pathway to promote cell proliferation and glycolysis in hepatocellular carcinoma cells. Pathway inhibition of MEK1/2-ERK1/2 reversed the effects of CENPL overexpression.","method":"CENPL overexpression and knockdown in HCC cells; MEK/ERK pathway inhibitor treatment; proliferation and glycolysis assays; western blotting for MAPK pathway components","journal":"The International Journal of Biochemistry & Cell Biology","confidence":"Low","confidence_rationale":"Tier 3 / Weak — pharmacological inhibitor plus OE/KD, single lab, no direct demonstration of CENPL-MEK interaction","pmids":["37914022"],"is_preprint":false},{"year":2024,"finding":"KDM1A promotes epithelial ovarian cancer progression through a KDM1A→ZFP64→CENPL transcriptional axis: KDM1A demethylates H3K9me2 at the ZFP64 promoter to activate ZFP64, which in turn activates CENPL transcription. CENPL re-expression rescued tumor growth suppressed by ZFP64 knockdown in vivo.","method":"Chromatin immunoprecipitation (ChIP) for H3K9me2 at ZFP64 promoter; knockdown/overexpression epistasis; in vivo xenograft rescue experiment","journal":"Cytotechnology","confidence":"Medium","confidence_rationale":"Tier 2 / Weak — ChIP for epigenetic mark plus in vivo epistasis rescue, single lab, establishes transcriptional regulation of CENPL","pmids":["39628712"],"is_preprint":false}],"current_model":"CENPL encodes a constitutive centromere-associated network (CCAN) protein that is recruited to CENP-A nucleosomes as a distal component of the NAC, where it acts as a scaffold bridging CENP-N to the CENP-PORQU sub-complex and CENP-K; CENPL is essential for kinetochore microtubule stability, poleward flux, and the kinetochore pushing force that ensures faithful amphitelic attachment, and its centromere localization is dynamically regulated by cell cycle-dependent phosphorylation of the CENP-LN complex; beyond its canonical kinetochore role, CENPL has been placed downstream of the E2F8 transcription factor in homologous recombination repair and chemoresistance, and downstream of a KDM1A-ZFP64 epigenetic axis in cancer contexts."},"narrative":{"mechanistic_narrative":"CENPL encodes a constitutive centromere-associated network (CCAN) protein recruited to CENP-A nucleosomes as a CENP-A-nucleosome distal component of the NAC, where its disruption causes chromosome alignment and segregation failures incompatible with cell survival [PMID:16622419]. CENPL functions as a scaffolding hub: it binds CENP-N and, together with CENP-K, mediates docking of the CENP-PORQU sub-complex to the CCAN [PMID:23028590], and it co-migrates with CENP-K within a soluble nucleoplasmic CCAN complex outside centromeres [PMID:29509805]. At the kinetochore, CENPL is required for kinetochore microtubule stability, homogeneous poleward microtubule flux, and the kinetochore pushing force driving centrosome separation, such that its loss produces syntelic and merotelic attachments [PMID:22399803]. Recruitment of the CENP-LN module is gated by cell cycle-dependent phosphorylation of both CENPL and CENP-N, with cycles of phosphorylation and dephosphorylation required for proper centromere localization and CCAN reorganization [PMID:35830614]. Beyond its kinetochore role, CENPL is a transcriptional target acting downstream of E2F8 in homologous recombination repair of double-strand breaks and chemoresistance [PMID:38522807] and downstream of a KDM1A→ZFP64 epigenetic axis in cancer [PMID:39628712].","teleology":[{"year":2006,"claim":"Established that CENPL is a bona fide centromere protein by placing it within the CENP-A nucleosome-associated complex and showing its loss abolishes faithful chromosome segregation.","evidence":"AP-MS of CENP-A nucleosome complexes plus RNAi depletion with mitotic phenotype readout in human cells","pmids":["16622419"],"confidence":"High","gaps":["Did not resolve which CCAN partners directly contact CENPL","Molecular basis of the segregation defect not defined"]},{"year":2012,"claim":"Defined the specific mechanical contribution of CENPL by linking it to kinetochore microtubule stability, flux, and pushing force, explaining why its loss yields erroneous attachments.","evidence":"siRNA depletion with live-cell imaging quantifying centrosome separation, microtubule flux, and attachment geometry in human cells","pmids":["22399803"],"confidence":"High","gaps":["Does not establish whether these defects are direct or downstream of CCAN disassembly","No structural mechanism for force generation"]},{"year":2012,"claim":"Positioned CENPL as a scaffold by mapping it as the CENP-N binding partner that, with CENP-K, recruits the CENP-PORQU sub-complex to the CCAN.","evidence":"Fluorescent three-hybrid, FRET, and co-immunoprecipitation interaction mapping in mammalian cells","pmids":["23028590"],"confidence":"Medium","gaps":["Interaction interfaces not resolved at residue level","Single lab; affinities not quantified"]},{"year":2013,"claim":"Provided structural and biochemical insight via the yeast ortholog, revealing an elongated Iml3 fold, heterodimerization with the Chl4 (CENP-N) C-terminus, and low-affinity DNA binding by the heterodimer.","evidence":"X-ray crystallography of Iml3, pull-down domain mapping, and in vitro DNA-binding assays in yeast","pmids":["24311582"],"confidence":"Medium","gaps":["Not directly demonstrated for human CENPL","Functional relevance of DNA binding in cells untested"]},{"year":2018,"claim":"Showed that CENPL exists in a preassembled soluble CCAN pool, indicating complex formation precedes centromere docking.","evidence":"Fluorescence cross-correlation spectroscopy in living human interphase cells","pmids":["29509805"],"confidence":"Medium","gaps":["Stoichiometry of the soluble complex not determined","Single method, single lab"]},{"year":2022,"claim":"Identified the regulatory logic of CENPL recruitment, showing phosphorylation cycles on CENPL and CENP-N control CENP-LN complex formation and centromere localization across the cell cycle.","evidence":"Phosphomimetic and phospho-null mutagenesis with live-cell imaging and chromosome segregation assays","pmids":["35830614"],"confidence":"High","gaps":["Responsible kinases and phosphatases not identified","Phosphosite occupancy in vivo not quantified"]},{"year":2023,"claim":"Reported a non-canonical role linking CENPL to MEK1/2-ERK1/2 signaling driving proliferation and glycolysis in hepatocellular carcinoma.","evidence":"Overexpression/knockdown plus MEK/ERK inhibitor treatment with proliferation and glycolysis assays in HCC cells","pmids":["37914022"],"confidence":"Low","gaps":["No direct CENPL-MEK interaction demonstrated; pharmacological inference only","Single lab, not independently confirmed","Mechanistic connection to kinetochore role unclear"]},{"year":2024,"claim":"Placed CENPL downstream of E2F8 in homologous recombination repair and chemoresistance via a rescue epistasis relationship.","evidence":"Knockdown/overexpression of CENPL and E2F8 with HR repair and chemosensitivity assays in breast cancer cells","pmids":["38522807"],"confidence":"Medium","gaps":["Molecular mechanism by which CENPL promotes HR not defined","Single lab"]},{"year":2024,"claim":"Established transcriptional control of CENPL by a KDM1A→ZFP64 epigenetic axis driving ovarian cancer growth.","evidence":"ChIP for H3K9me2 at the ZFP64 promoter, knockdown/overexpression epistasis, and in vivo xenograft rescue","pmids":["39628712"],"confidence":"Medium","gaps":["Direct ZFP64 binding at the CENPL promoter not shown","Downstream effector function of CENPL in this context unresolved"]},{"year":null,"claim":"How the kinetochore scaffolding role of CENPL mechanistically connects to its reported roles in HR repair, signaling, and cancer transcriptional programs remains unresolved.","evidence":"","pmids":[],"confidence":"Medium","gaps":["No unifying mechanism linking centromere function to DNA repair/proliferation roles","Kinases regulating CENP-LN phosphorylation unidentified","No human structural model of the CENPL-CENP-N interface"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,3]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0,1]},{"term_id":"GO:0005654","term_label":"nucleoplasm","supporting_discovery_ids":[3]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1,4]}],"complexes":["CENP-A nucleosome-associated complex (NAC/CCAN)","CENP-LN complex"],"partners":["CENPN","CENPK","CENPC","CENPH","CENPI","CENPM","CENPT","CENPW"],"other_free_text":[]}},"prefetch_data":{"uniprot":{"accession":"Q8N0S6","full_name":"Centromere protein L","aliases":["Interphase centromere complex protein 33"],"length_aa":344,"mass_kda":39.0,"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","subcellular_location":"Nucleus; Chromosome, centromere","url":"https://www.uniprot.org/uniprotkb/Q8N0S6/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CENPL","classification":"Common Essential","n_dependent_lines":588,"n_total_lines":1208,"dependency_fraction":0.4867549668874172},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CENPL","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":8.9},{"tissue":"testis","ntpm":9.1}],"url":"https://www.proteinatlas.org/search/CENPL"},"hgnc":{"alias_symbol":["dJ383J4.3","FLJ31044"],"prev_symbol":["C1orf155"]},"alphafold":{"accession":"Q8N0S6","domains":[{"cath_id":"-","chopping":"47-106_113-209","consensus_level":"high","plddt":88.2866,"start":47,"end":209},{"cath_id":"-","chopping":"214-316","consensus_level":"high","plddt":93.3078,"start":214,"end":316}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N0S6","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N0S6-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q8N0S6-F1-predicted_aligned_error_v6.png","plddt_mean":83.06},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CENPL","jax_strain_url":"https://www.jax.org/strain/search?query=CENPL"},"sequence":{"accession":"Q8N0S6","fasta_url":"https://rest.uniprot.org/uniprotkb/Q8N0S6.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q8N0S6/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q8N0S6"}},"corpus_meta":[{"pmid":"16622419","id":"PMC_16622419","title":"The human CENP-A centromeric nucleosome-associated complex.","date":"2006","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/16622419","citation_count":594,"is_preprint":false},{"pmid":"22399803","id":"PMC_22399803","title":"Kinetochores accelerate centrosome separation to ensure faithful chromosome segregation.","date":"2012","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/22399803","citation_count":39,"is_preprint":false},{"pmid":"21548129","id":"PMC_21548129","title":"Distinctive phenotype in 9 patients with deletion of chromosome 1q24-q25.","date":"2011","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/21548129","citation_count":34,"is_preprint":false},{"pmid":"23028590","id":"PMC_23028590","title":"Step-wise assembly, maturation and dynamic behavior of the human CENP-P/O/R/Q/U kinetochore sub-complex.","date":"2012","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/23028590","citation_count":33,"is_preprint":false},{"pmid":"24402816","id":"PMC_24402816","title":"Identifying novel oncogenes: a machine learning approach.","date":"2014","source":"Interdisciplinary sciences, computational life sciences","url":"https://pubmed.ncbi.nlm.nih.gov/24402816","citation_count":17,"is_preprint":false},{"pmid":"24311370","id":"PMC_24311370","title":"Pituitary deficiency and congenital infiltrating lipomatosis of the face in a girl with deletion of chromosome 1q24.3q31.1.","date":"2013","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/24311370","citation_count":14,"is_preprint":false},{"pmid":"35830614","id":"PMC_35830614","title":"Dynamic cell cycle-dependent phosphorylation modulates CENP-L-CENP-N centromere recruitment.","date":"2022","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/35830614","citation_count":13,"is_preprint":false},{"pmid":"29509805","id":"PMC_29509805","title":"CENP-C/H/I/K/M/T/W/N/L and hMis12 but not CENP-S/X participate in complex formation in the nucleoplasm of living human interphase cells outside centromeres.","date":"2018","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/29509805","citation_count":11,"is_preprint":false},{"pmid":"34012666","id":"PMC_34012666","title":"The roles of the cell division cycle-associated gene family in hepatocellular carcinoma.","date":"2021","source":"Journal of gastrointestinal oncology","url":"https://pubmed.ncbi.nlm.nih.gov/34012666","citation_count":9,"is_preprint":false},{"pmid":"38522807","id":"PMC_38522807","title":"E2F8-CENPL pathway contributes to homologous recombination repair and chemoresistance in breast cancer.","date":"2024","source":"Cellular signalling","url":"https://pubmed.ncbi.nlm.nih.gov/38522807","citation_count":8,"is_preprint":false},{"pmid":"24311582","id":"PMC_24311582","title":"Structural insights into the role of the Chl4-Iml3 complex in kinetochore assembly.","date":"2013","source":"Acta crystallographica. Section D, Biological crystallography","url":"https://pubmed.ncbi.nlm.nih.gov/24311582","citation_count":8,"is_preprint":false},{"pmid":"37914022","id":"PMC_37914022","title":"CENPL accelerates cell proliferation, cell cycle, apoptosis, and glycolysis via the MEK1/2-ERK1/2 pathway in hepatocellular carcinoma.","date":"2023","source":"The international journal of biochemistry & cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/37914022","citation_count":6,"is_preprint":false},{"pmid":"27416976","id":"PMC_27416976","title":"Nine year old boy with chromosome 1q23.3-q25.1 deletion.","date":"2016","source":"American journal of medical genetics. Part A","url":"https://pubmed.ncbi.nlm.nih.gov/27416976","citation_count":5,"is_preprint":false},{"pmid":"574496","id":"PMC_574496","title":"A cytogenetic survey of an institution for the metnally retarded. III. Q-Band chromosome heteromorphisms.","date":"1979","source":"Human genetics","url":"https://pubmed.ncbi.nlm.nih.gov/574496","citation_count":5,"is_preprint":false},{"pmid":"37605944","id":"PMC_37605944","title":"Delineation of the CENP-LN sub-complex dissociation mechanism upon multisite phosphorylation during mitosis.","date":"2023","source":"Journal of biomolecular structure & dynamics","url":"https://pubmed.ncbi.nlm.nih.gov/37605944","citation_count":4,"is_preprint":false},{"pmid":"39628712","id":"PMC_39628712","title":"KDM1A-mediated ZFP64 demethylation activates CENPL to promote epithelial ovarian cancer progression.","date":"2024","source":"Cytotechnology","url":"https://pubmed.ncbi.nlm.nih.gov/39628712","citation_count":1,"is_preprint":false},{"pmid":"41751958","id":"PMC_41751958","title":"FOXM1 Signaling Network Transcriptionally Upregulates Expression of Proteins Involved in Mitotic Progression to Induce High Proliferation and Chromosomal Instability in Androgen Receptor-Low Triple-Negative Breast Cancer.","date":"2026","source":"International journal of molecular sciences","url":"https://pubmed.ncbi.nlm.nih.gov/41751958","citation_count":1,"is_preprint":false},{"pmid":"40067534","id":"PMC_40067534","title":"Towards identification of a holocentromere marker in the lepidopteran model Spodoptera frugiperda.","date":"2025","source":"Chromosoma","url":"https://pubmed.ncbi.nlm.nih.gov/40067534","citation_count":0,"is_preprint":false},{"pmid":"41927050","id":"PMC_41927050","title":"Functional characterization of porcine septin12 and its role in male reproduction.","date":"2026","source":"Animal bioscience","url":"https://pubmed.ncbi.nlm.nih.gov/41927050","citation_count":0,"is_preprint":false},{"pmid":"33062452","id":"PMC_33062452","title":"Loss of inner kinetochore genes is associated with the transition to an unconventional point centromere in budding yeast.","date":"2020","source":"PeerJ","url":"https://pubmed.ncbi.nlm.nih.gov/33062452","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":11579,"output_tokens":2469,"usd":0.035886,"stage1_stop_reason":"end_turn"},"stage2":{"model":"claude-opus-4-8","input_tokens":9555,"output_tokens":2868,"usd":0.059737,"stage2_stop_reason":"end_turn"},"total_usd":0.095623,"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\": 2006,\n      \"finding\": \"CENP-L (CENPL) is a component of the CENP-A nucleosome-associated complex (NAC) at human centromeres. CENP-A nucleosomes directly recruit the NAC, and CENP-L is identified as one of seven CENP-A-nucleosome distal (CAD) components assembling on the NAC. Disruption of the NAC causes chromosome alignment and segregation errors that preclude cell survival.\",\n      \"method\": \"Affinity purification / mass spectrometry (AP-MS) of CENP-A nucleosome complexes; RNAi depletion with mitotic phenotype readout\",\n      \"journal\": \"Nature Cell Biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Strong — reciprocal affinity purification with MS identification, functional depletion experiments, replicated by subsequent studies\",\n      \"pmids\": [\"16622419\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"CENPL is essential for the stability of kinetochore microtubules, for a homogeneous poleward microtubule flux rate, and for the kinetochore pushing force that accelerates centrosome separation in prometaphase. CENP-L depletion causes massive chromosome alignment and segregation defects due to syntelic and merotelic kinetochore-microtubule attachments.\",\n      \"method\": \"siRNA depletion of CENP-L in human cells; live-cell imaging and quantitative measurement of centrosome separation kinetics, microtubule flux, and kinetochore-microtubule attachment geometry\",\n      \"journal\": \"Journal of Cell Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — clean KO/KD with defined cellular phenotypes (microtubule stability, flux, attachment fidelity), multiple orthogonal readouts in a single study\",\n      \"pmids\": [\"22399803\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2012,\n      \"finding\": \"CENP-P/O/R/Q/U sub-complex binding to the CCAN is largely mediated through interactions with CENP-L (as the CENP-N binding partner) and CENP-K. CENP-L thus acts as a hub connecting the CENP-N module to the CENP-PORQU sub-complex.\",\n      \"method\": \"Fluorescent three-hybrid (F3H) assay, FRET in living mammalian cells, and co-immunoprecipitation to map protein-protein interactions\",\n      \"journal\": \"PloS One\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Moderate — multiple orthogonal interaction assays (F3H, FRET, Co-IP) in a single study, single lab\",\n      \"pmids\": [\"23028590\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2018,\n      \"finding\": \"CENP-L co-migrates with CENP-K in the nucleoplasm outside centromeres in living human interphase cells, demonstrating that CENP-L and CENP-K form part of a soluble CENP-C/H/I/K/M/T/W/N/L complex in the nucleoplasm independently of centromere binding.\",\n      \"method\": \"Fluorescence cross-correlation spectroscopy (FCCS) in living human interphase cells\",\n      \"journal\": \"PloS One\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — direct in-cell biophysical measurement (FCCS) establishing co-migration, single lab, single method\",\n      \"pmids\": [\"29509805\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"Phosphorylation of CENP-L and CENP-N controls CENP-LN complex formation and centromere localization in a cell cycle-dependent manner. Mimicking constitutive phosphorylation of either CENP-L or CENP-N, or simultaneously preventing phosphorylation of both, prevents CENP-LN localization and disrupts chromosome segregation. Cycles of phosphorylation and dephosphorylation are critical for CENP-LN complex recruitment and CCAN reorganization across the cell cycle.\",\n      \"method\": \"Phosphomimetic and phospho-null mutagenesis of CENP-L and CENP-N; live-cell imaging; chromosome segregation assays\",\n      \"journal\": \"Molecular Biology of the Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 / Moderate — direct mutagenesis of phosphosites with functional readouts (localization and chromosome segregation), multiple mutant conditions tested, single lab\",\n      \"pmids\": [\"35830614\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2013,\n      \"finding\": \"The yeast CENP-L ortholog Iml3 adopts an elongated conformation with intramolecular interactions (crystal structure resolved). Its binding partner Chl4 (CENP-N ortholog) C-terminal domain forms a dimer and is responsible for Iml3 binding. The Chl4-Iml3 heterodimer exhibits low-affinity nonspecific DNA-binding activity, potentially relevant to kinetochore assembly.\",\n      \"method\": \"X-ray crystallography of Iml3; pull-down assays mapping Chl4-Iml3 interaction domains; in vitro DNA-binding assays\",\n      \"journal\": \"Acta Crystallographica Section D\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1 / Weak — crystal structure and in vitro biochemistry for the yeast ortholog, not directly demonstrated for human CENPL\",\n      \"pmids\": [\"24311582\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"CENPL is a transcriptional target of E2F8 and mediates E2F8's role in homologous recombination (HR) repair of DNA double-strand breaks and chemoresistance. CENPL overexpression in E2F8-knockdown cells partially rescued HR repair and chemotherapy sensitivity; CENPL knockdown alone impaired HR repair and sensitized cells to DNA-damaging drugs.\",\n      \"method\": \"siRNA knockdown and overexpression of CENPL and E2F8; HR repair assays; chemosensitivity assays in breast cancer cells; transcriptional target validation\",\n      \"journal\": \"Cellular Signalling\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — rescue epistasis experiment placing CENPL downstream of E2F8 in HR repair, single lab, multiple functional assays\",\n      \"pmids\": [\"38522807\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CENPL activates the MEK1/2-ERK1/2 signaling pathway to promote cell proliferation and glycolysis in hepatocellular carcinoma cells. Pathway inhibition of MEK1/2-ERK1/2 reversed the effects of CENPL overexpression.\",\n      \"method\": \"CENPL overexpression and knockdown in HCC cells; MEK/ERK pathway inhibitor treatment; proliferation and glycolysis assays; western blotting for MAPK pathway components\",\n      \"journal\": \"The International Journal of Biochemistry & Cell Biology\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 / Weak — pharmacological inhibitor plus OE/KD, single lab, no direct demonstration of CENPL-MEK interaction\",\n      \"pmids\": [\"37914022\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"KDM1A promotes epithelial ovarian cancer progression through a KDM1A→ZFP64→CENPL transcriptional axis: KDM1A demethylates H3K9me2 at the ZFP64 promoter to activate ZFP64, which in turn activates CENPL transcription. CENPL re-expression rescued tumor growth suppressed by ZFP64 knockdown in vivo.\",\n      \"method\": \"Chromatin immunoprecipitation (ChIP) for H3K9me2 at ZFP64 promoter; knockdown/overexpression epistasis; in vivo xenograft rescue experiment\",\n      \"journal\": \"Cytotechnology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 / Weak — ChIP for epigenetic mark plus in vivo epistasis rescue, single lab, establishes transcriptional regulation of CENPL\",\n      \"pmids\": [\"39628712\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CENPL encodes a constitutive centromere-associated network (CCAN) protein that is recruited to CENP-A nucleosomes as a distal component of the NAC, where it acts as a scaffold bridging CENP-N to the CENP-PORQU sub-complex and CENP-K; CENPL is essential for kinetochore microtubule stability, poleward flux, and the kinetochore pushing force that ensures faithful amphitelic attachment, and its centromere localization is dynamically regulated by cell cycle-dependent phosphorylation of the CENP-LN complex; beyond its canonical kinetochore role, CENPL has been placed downstream of the E2F8 transcription factor in homologous recombination repair and chemoresistance, and downstream of a KDM1A-ZFP64 epigenetic axis in cancer contexts.\"\n}\n```","stage2_raw":"{\n  \"mechanistic_narrative\": \"CENPL encodes a constitutive centromere-associated network (CCAN) protein recruited to CENP-A nucleosomes as a CENP-A-nucleosome distal component of the NAC, where its disruption causes chromosome alignment and segregation failures incompatible with cell survival [#0]. CENPL functions as a scaffolding hub: it binds CENP-N and, together with CENP-K, mediates docking of the CENP-PORQU sub-complex to the CCAN [#2], and it co-migrates with CENP-K within a soluble nucleoplasmic CCAN complex outside centromeres [#3]. At the kinetochore, CENPL is required for kinetochore microtubule stability, homogeneous poleward microtubule flux, and the kinetochore pushing force driving centrosome separation, such that its loss produces syntelic and merotelic attachments [#1]. Recruitment of the CENP-LN module is gated by cell cycle-dependent phosphorylation of both CENPL and CENP-N, with cycles of phosphorylation and dephosphorylation required for proper centromere localization and CCAN reorganization [#4]. Beyond its kinetochore role, CENPL is a transcriptional target acting downstream of E2F8 in homologous recombination repair of double-strand breaks and chemoresistance [#6] and downstream of a KDM1A\\u2192ZFP64 epigenetic axis in cancer [#8].\",\n  \"teleology\": [\n    {\n      \"year\": 2006,\n      \"claim\": \"Established that CENPL is a bona fide centromere protein by placing it within the CENP-A nucleosome-associated complex and showing its loss abolishes faithful chromosome segregation.\",\n      \"evidence\": \"AP-MS of CENP-A nucleosome complexes plus RNAi depletion with mitotic phenotype readout in human cells\",\n      \"pmids\": [\"16622419\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Did not resolve which CCAN partners directly contact CENPL\", \"Molecular basis of the segregation defect not defined\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Defined the specific mechanical contribution of CENPL by linking it to kinetochore microtubule stability, flux, and pushing force, explaining why its loss yields erroneous attachments.\",\n      \"evidence\": \"siRNA depletion with live-cell imaging quantifying centrosome separation, microtubule flux, and attachment geometry in human cells\",\n      \"pmids\": [\"22399803\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Does not establish whether these defects are direct or downstream of CCAN disassembly\", \"No structural mechanism for force generation\"]\n    },\n    {\n      \"year\": 2012,\n      \"claim\": \"Positioned CENPL as a scaffold by mapping it as the CENP-N binding partner that, with CENP-K, recruits the CENP-PORQU sub-complex to the CCAN.\",\n      \"evidence\": \"Fluorescent three-hybrid, FRET, and co-immunoprecipitation interaction mapping in mammalian cells\",\n      \"pmids\": [\"23028590\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Interaction interfaces not resolved at residue level\", \"Single lab; affinities not quantified\"]\n    },\n    {\n      \"year\": 2013,\n      \"claim\": \"Provided structural and biochemical insight via the yeast ortholog, revealing an elongated Iml3 fold, heterodimerization with the Chl4 (CENP-N) C-terminus, and low-affinity DNA binding by the heterodimer.\",\n      \"evidence\": \"X-ray crystallography of Iml3, pull-down domain mapping, and in vitro DNA-binding assays in yeast\",\n      \"pmids\": [\"24311582\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Not directly demonstrated for human CENPL\", \"Functional relevance of DNA binding in cells untested\"]\n    },\n    {\n      \"year\": 2018,\n      \"claim\": \"Showed that CENPL exists in a preassembled soluble CCAN pool, indicating complex formation precedes centromere docking.\",\n      \"evidence\": \"Fluorescence cross-correlation spectroscopy in living human interphase cells\",\n      \"pmids\": [\"29509805\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Stoichiometry of the soluble complex not determined\", \"Single method, single lab\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Identified the regulatory logic of CENPL recruitment, showing phosphorylation cycles on CENPL and CENP-N control CENP-LN complex formation and centromere localization across the cell cycle.\",\n      \"evidence\": \"Phosphomimetic and phospho-null mutagenesis with live-cell imaging and chromosome segregation assays\",\n      \"pmids\": [\"35830614\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Responsible kinases and phosphatases not identified\", \"Phosphosite occupancy in vivo not quantified\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"Reported a non-canonical role linking CENPL to MEK1/2-ERK1/2 signaling driving proliferation and glycolysis in hepatocellular carcinoma.\",\n      \"evidence\": \"Overexpression/knockdown plus MEK/ERK inhibitor treatment with proliferation and glycolysis assays in HCC cells\",\n      \"pmids\": [\"37914022\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No direct CENPL-MEK interaction demonstrated; pharmacological inference only\", \"Single lab, not independently confirmed\", \"Mechanistic connection to kinetochore role unclear\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Placed CENPL downstream of E2F8 in homologous recombination repair and chemoresistance via a rescue epistasis relationship.\",\n      \"evidence\": \"Knockdown/overexpression of CENPL and E2F8 with HR repair and chemosensitivity assays in breast cancer cells\",\n      \"pmids\": [\"38522807\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Molecular mechanism by which CENPL promotes HR not defined\", \"Single lab\"]\n    },\n    {\n      \"year\": 2024,\n      \"claim\": \"Established transcriptional control of CENPL by a KDM1A\\u2192ZFP64 epigenetic axis driving ovarian cancer growth.\",\n      \"evidence\": \"ChIP for H3K9me2 at the ZFP64 promoter, knockdown/overexpression epistasis, and in vivo xenograft rescue\",\n      \"pmids\": [\"39628712\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Direct ZFP64 binding at the CENPL promoter not shown\", \"Downstream effector function of CENPL in this context unresolved\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"How the kinetochore scaffolding role of CENPL mechanistically connects to its reported roles in HR repair, signaling, and cancer transcriptional programs remains unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No unifying mechanism linking centromere function to DNA repair/proliferation roles\", \"Kinases regulating CENP-LN phosphorylation unidentified\", \"No human structural model of the CENPL-CENP-N interface\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 3]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"GO:0005654\", \"supporting_discovery_ids\": [3]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1, 4]}\n    ],\n    \"complexes\": [\"CENP-A nucleosome-associated complex (NAC/CCAN)\", \"CENP-LN complex\"],\n    \"partners\": [\"CENPN\", \"CENPK\", \"CENPC\", \"CENPH\", \"CENPI\", \"CENPM\", \"CENPT\", \"CENPW\"],\n    \"other_free_text\": []\n  }\n}","audit_flag":null,"evaluation":{"pairwise":"win","faith_supported":5,"faith_total":5,"faith_pct":100.0}}