{"gene":"CENPI","run_date":"2026-04-28T17:28:52","timeline":{"discoveries":[{"year":2003,"finding":"Human CENP-I is required for the localization of CENP-F and the spindle checkpoint proteins MAD1 and MAD2 to kinetochores; depletion of CENP-I causes G2 delay and inability to arrest in mitosis despite unattached kinetochores, with residual mitotic delay being MAD2-dependent and requiring collective signal from many unattached kinetochores.","method":"RNAi depletion, immunofluorescence, checkpoint analysis in human cells","journal":"Nature cell biology","confidence":"High","confidence_rationale":"Tier 2 — clean depletion with defined cellular phenotypes, multiple orthogonal readouts, replicated concept across studies","pmids":["12640463"],"is_preprint":false},{"year":2002,"finding":"CENP-I is a constitutive centromere component that colocalizes with CENP-A, -C, and -H throughout the cell cycle; loss of CENP-I in chicken DT40 cells causes prometaphase arrest with misaligned chromosomes; both CENP-I and CENP-H are required for localization of CENP-C but not CENP-A to the centromere.","method":"Conditional knockout in chicken DT40 cells, immunocytochemistry, co-localization analysis","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 — conditional loss-of-function with defined molecular hierarchy, replicated in vertebrate model","pmids":["11970896"],"is_preprint":false},{"year":2014,"finding":"CENP-I is required to generate stable association of the RZZ complex and Mad1 with kinetochores, and also inhibits their dynein-mediated stripping; Aurora B regulates RZZ/Mad1 association while CENP-I inhibits dissociation, together forming a molecular switch maintaining spindle checkpoint signal at prometaphase kinetochores until mature microtubule attachment.","method":"CENP-I depletion (RNAi), Aurora B inhibition, immunofluorescence, epistasis analysis in human cells","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 2 — epistasis between CENP-I and Aurora B established with multiple orthogonal methods, clear mechanistic model","pmids":["24862574"],"is_preprint":false},{"year":2015,"finding":"CENP-I can recruit M18BP1 (Mis18BP1) and enhance its assembly at centromeres downstream of CENP-C, functioning as a key connecting factor between kinetochore structure and CENP-A assembly; tethering CENP-I to ectopic sites induced de novo CENP-A assembly by recruiting CENP-C and subsequently M18BP1.","method":"Tetracycline repressor tethering to synthetic alphoid DNA arrays in human artificial chromosomes, immunofluorescence","journal":"Journal of cell science","confidence":"Medium","confidence_rationale":"Tier 2 — direct tethering experiment with functional readout, single study","pmids":["26527398"],"is_preprint":false},{"year":2023,"finding":"CENP-I directly interacts with centromeric DNA, preferentially recognizing AT-rich elements via a consecutive DNA-binding surface formed by conserved charged residues at the end of N-terminal HEAT repeats; DNA-binding-deficient mutants retain interaction with CENP-H/K and CENP-M but show significantly diminished centromeric localization, impaired chromosome alignment, and reduced centromeric loading of newly synthesized CENP-A; CENP-I stabilizes CENP-A nucleosomes by binding nucleosomal DNA rather than histones.","method":"In vitro DNA-binding assays, mutagenesis, co-immunoprecipitation, immunofluorescence, CENP-A loading assays in human cells","journal":"Proceedings of the National Academy of Sciences of the United States of America","confidence":"High","confidence_rationale":"Tier 1 — in vitro reconstitution with mutagenesis, structural surface mapping, and functional cellular validation","pmids":["36888657"],"is_preprint":false},{"year":2020,"finding":"A conserved helix (α11) in CENP-I forms intramolecular interactions with N-terminal HEAT repeats; deletion of this helix in human CENP-I dramatically reduces interaction with CENP-H and CENP-M, and mutations of conserved residues on the helix specifically weaken binding to CENP-M but not CENP-H in HeLa cells.","method":"Structural analysis, in vitro aggregation assays, co-immunoprecipitation in HeLa cells, mutagenesis","journal":"Journal of molecular recognition","confidence":"Medium","confidence_rationale":"Tier 1-2 — mutagenesis with in vitro and cellular binding assays, single study","pmids":["32017295"],"is_preprint":false},{"year":2021,"finding":"A conserved surface of Ctf3/CENP-I (budding yeast ortholog) provides a direct binding site for the desumoylase Ulp2; Ctf3 mutations disabling Ulp2 recruitment cause elevated inner kinetochore sumoylation and defective chromosome segregation.","method":"Structural analysis, mutagenesis, yeast genetics, sumoylation assays, chromosome segregation assays","journal":"The Journal of cell biology","confidence":"High","confidence_rationale":"Tier 1-2 — structural surface identified, mutagenesis with functional readout, ortholog of CENP-I","pmids":["34081091"],"is_preprint":false},{"year":2021,"finding":"Loss of CENP-I impairs homologous recombination (HR) DSB repair while having no effect on non-homologous end-joining; CENP-I deficiency increases R-loop formation, and RNaseH1 expression restores HR capacity in CENP-I-deficient cells, linking CENP-I to R-loop resolution as part of its HR function.","method":"CENP-I knockout/knockdown, HR and NHEJ reporter assays, 53BP1 foci analysis, RNaseH1 rescue experiments, PARP inhibitor sensitivity assay","journal":"Cancers","confidence":"Medium","confidence_rationale":"Tier 2 — multiple orthogonal assays establishing HR role, single study","pmids":["34206916"],"is_preprint":false},{"year":2022,"finding":"The inner kinetochore protein Mis6 (CENP-I ortholog in fission yeast) and Mis15 (CENP-N) retain pre-existing CENP-A at centromeres during mitosis by blocking non-coding RNA transcription at the central core centromere region; eliminating Mis6 during mitosis caused immediate loss of CENP-A due to upregulation of non-coding RNAs, and RNA polymerase II inhibition prevented CENP-A loss in mis6 mutants.","method":"Conditional depletion in fission yeast, immunofluorescence, RNA analysis, RNA Pol II inhibition rescue experiment","journal":"Communications biology","confidence":"Medium","confidence_rationale":"Tier 2 — genetic epistasis with pharmacological rescue, fission yeast ortholog, single study","pmids":["35970865"],"is_preprint":false},{"year":2002,"finding":"Budding yeast Ctf3p (CENP-I ortholog) interacts with Mcm22p and Mcm16p at the outer kinetochore, and all three bind centromere DNA in a Ctf19p-dependent manner as shown by chromatin immunoprecipitation; unlike fission yeast Mis6, Ctf3p is not required for loading of the CENP-A homolog Cse4p, but Ctf3p and Ctf19p fail to bind centromere DNA in a cse4-1 mutant.","method":"Synthetic dosage lethality screen, two-hybrid interaction, chromatin immunoprecipitation, genetic epistasis in budding yeast","journal":"Genes & development","confidence":"High","confidence_rationale":"Tier 2 — ChIP plus genetic epistasis, multiple orthogonal methods, foundational study","pmids":["11782448"],"is_preprint":false},{"year":2000,"finding":"The fission yeast CENP-A homolog (SpCENP-A) binding to inner centromeres depends on Mis6 (CENP-I ortholog); Mis6 acts during G1-S phase and is required for recruiting SpCENP-A to form proper centromere chromatin for chromosome segregation.","method":"Temperature-sensitive mutant analysis, genetic epistasis, cell cycle staging, chromosome segregation assays in fission yeast","journal":"Science","confidence":"High","confidence_rationale":"Tier 2 — epistasis with cell cycle resolution, highly cited foundational study","pmids":["10864871"],"is_preprint":false},{"year":2005,"finding":"The Mis6-complex (containing fission yeast CENP-I ortholog) physically interacts with Mad2 under conditions where the Mad2-dependent checkpoint is activated, and is required for accumulation of Mad2 onto unattached kinetochores; the Mis6-complex also collaborates with the Nuf2-complex to monitor spindle-kinetochore attachment state.","method":"Co-immunoprecipitation, immunofluorescence, genetic analysis of kinetochore components in fission yeast","journal":"Molecular biology of the cell","confidence":"Medium","confidence_rationale":"Tier 2-3 — direct interaction by co-IP plus genetic requirement, fission yeast ortholog, single study","pmids":["15930132"],"is_preprint":false}],"current_model":"CENP-I is a constitutive inner kinetochore protein that directly binds AT-rich centromeric DNA via its N-terminal HEAT repeats to stabilize CENP-A nucleosomes and promote new CENP-A loading, recruits M18BP1 downstream of CENP-C to maintain centromere identity, is required for localization of CENP-C, CENP-F, MAD1, and MAD2 to kinetochores, acts together with Aurora B as a molecular switch to sustain spindle checkpoint signaling by stabilizing RZZ/Mad1 at unattached kinetochores and inhibiting dynein-mediated stripping, plays a role in homologous recombination DSB repair through R-loop suppression, and in yeast its ortholog docks the desumoylase Ulp2 to regulate kinetochore sumoylation."},"narrative":{"teleology":[{"year":2000,"claim":"The first functional link between a CENP-I ortholog and centromere chromatin was established when fission yeast Mis6 was shown to be required during G1-S for loading the CENP-A homolog, defining CENP-I as upstream of centromeric nucleosome specification.","evidence":"Temperature-sensitive mis6 mutant analysis with cell-cycle staging and chromosome segregation assays in S. pombe","pmids":["10864871"],"confidence":"High","gaps":["Mechanism by which Mis6 promotes CENP-A incorporation was unknown","Whether the requirement was direct or indirect was unresolved","Applicability to vertebrate centromeres not yet tested"]},{"year":2002,"claim":"Parallel work in budding yeast (Ctf3p) and chicken DT40 cells placed CENP-I as a constitutive centromere component required for CENP-C localization and proper chromosome segregation, revealing a conserved kinetochore assembly hierarchy in which CENP-I and CENP-H act upstream of CENP-C but downstream of CENP-A.","evidence":"Conditional knockout in DT40 cells with co-localization analysis; ChIP and genetic epistasis in budding yeast","pmids":["11970896","11782448"],"confidence":"High","gaps":["Whether CENP-I contacts centromeric DNA directly was unknown","Relationship between CENP-I and outer kinetochore assembly undefined in vertebrates"]},{"year":2003,"claim":"Human CENP-I was shown to be required for kinetochore recruitment of CENP-F, MAD1, and MAD2, establishing its role as a platform for spindle assembly checkpoint signaling in human cells.","evidence":"RNAi depletion of CENP-I in human cells with immunofluorescence and checkpoint analysis","pmids":["12640463"],"confidence":"High","gaps":["Whether CENP-I directly interacts with checkpoint proteins or acts indirectly was unclear","Mechanism by which residual MAD2-dependent delay persists after CENP-I depletion not resolved"]},{"year":2005,"claim":"Physical interaction between the Mis6 complex (CENP-I ortholog) and Mad2 was demonstrated in fission yeast, providing evidence that the inner kinetochore directly engages checkpoint machinery rather than simply serving as a structural scaffold.","evidence":"Co-immunoprecipitation and genetic analysis in S. pombe under checkpoint-activating conditions","pmids":["15930132"],"confidence":"Medium","gaps":["Whether the interaction is direct or bridged by other kinetochore components was not determined","Co-IP performed under one condition; generality across checkpoint states untested"]},{"year":2014,"claim":"The checkpoint role of CENP-I was refined to a dual function: generating stable RZZ/Mad1 association at kinetochores and inhibiting dynein-mediated stripping, working together with Aurora B as a molecular switch that sustains checkpoint signaling until mature microtubule attachment.","evidence":"Epistasis between CENP-I depletion and Aurora B inhibition in human cells with quantitative kinetochore localization analysis","pmids":["24862574"],"confidence":"High","gaps":["Direct biochemical basis for CENP-I inhibition of dynein-mediated stripping was not identified","How attachment status feeds back to relieve CENP-I's inhibitory role remained unknown"]},{"year":2015,"claim":"CENP-I was linked to centromere maintenance by its ability to recruit M18BP1 and induce de novo CENP-A assembly downstream of CENP-C, revealing it as a coupling factor between kinetochore structure and epigenetic centromere propagation.","evidence":"Ectopic tethering of CENP-I to synthetic alphoid arrays on human artificial chromosomes with immunofluorescence readout","pmids":["26527398"],"confidence":"Medium","gaps":["Physiological relevance of ectopic tethering system to endogenous centromeres not fully validated","Whether CENP-I directly contacts M18BP1 or acts through intermediate factors was not resolved"]},{"year":2020,"claim":"Structural dissection of a conserved intramolecular helix (α11) in CENP-I revealed that it mediates assembly with CENP-H and CENP-M through distinct binding interfaces, beginning to define the structural basis of the CENP-H/I/K/M sub-complex.","evidence":"Mutagenesis, in vitro aggregation assays, and co-immunoprecipitation in HeLa cells","pmids":["32017295"],"confidence":"Medium","gaps":["No high-resolution structure of the CENP-H/I/K/M complex was available","In vivo consequences of helix mutations on kinetochore function not tested"]},{"year":2021,"claim":"Two distinct non-kinetochore functions were uncovered: in budding yeast, a conserved surface of Ctf3/CENP-I docks the desumoylase Ulp2 to regulate inner kinetochore sumoylation; in human cells, CENP-I promotes homologous recombination DSB repair by suppressing R-loop accumulation.","evidence":"Structural analysis and mutagenesis with sumoylation/segregation assays in yeast; HR/NHEJ reporter assays and RNaseH1 rescue in human cells","pmids":["34081091","34206916"],"confidence":"High","gaps":["Whether the Ulp2-docking function is conserved in human CENP-I is unknown","Mechanism by which CENP-I suppresses R-loops (direct RNA binding vs indirect) was not determined","Whether the HR function operates at centromeric or genome-wide loci is unclear"]},{"year":2022,"claim":"Fission yeast Mis6 (CENP-I) was shown to retain pre-existing CENP-A during mitosis by blocking non-coding RNA transcription at centromeres, providing a mechanistic explanation for CENP-A stabilization distinct from direct DNA binding.","evidence":"Conditional depletion of Mis6 in mitotic S. pombe cells with RNA analysis and RNA Pol II inhibition rescue","pmids":["35970865"],"confidence":"Medium","gaps":["Whether this transcription-blocking mechanism operates in human cells is untested","Relationship between this RNA-based mechanism and direct DNA-binding stabilization of CENP-A is unresolved"]},{"year":2023,"claim":"The long-standing question of whether CENP-I contacts centromeric DNA directly was resolved: CENP-I binds AT-rich DNA through conserved charged residues at the end of N-terminal HEAT repeats, and this interaction is required for centromeric localization, chromosome alignment, and CENP-A loading — independent of CENP-H/K/M binding.","evidence":"In vitro DNA-binding assays, charge-reversal mutagenesis, co-immunoprecipitation, and CENP-A loading assays in human cells","pmids":["36888657"],"confidence":"High","gaps":["No atomic-resolution structure of CENP-I bound to nucleosomal DNA exists","Whether DNA-binding and transcription-blocking are the same or separable mechanisms is unclear"]},{"year":null,"claim":"A high-resolution structure of CENP-I bound to centromeric nucleosomal DNA is lacking, and the molecular basis by which CENP-I inhibits dynein-mediated stripping of checkpoint proteins, the conservation of the Ulp2-docking function in vertebrates, and the mechanism linking CENP-I to R-loop suppression remain unresolved.","evidence":"","pmids":[],"confidence":"Low","gaps":["No cryo-EM or crystal structure of CENP-I on nucleosomal DNA","Dynein-stripping inhibition mechanism not biochemically defined","R-loop suppression mechanism (direct vs indirect) uncharacterized"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0003677","term_label":"DNA binding","supporting_discovery_ids":[4,9]},{"term_id":"GO:0098772","term_label":"molecular function regulator activity","supporting_discovery_ids":[2,6]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0,1,4,9]},{"term_id":"GO:0005634","term_label":"nucleus","supporting_discovery_ids":[7]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[0,1,2,10]},{"term_id":"R-HSA-73894","term_label":"DNA Repair","supporting_discovery_ids":[7]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[4,8,10]}],"complexes":["CENP-H/I/K/M complex","Constitutive centromere-associated network (CCAN)"],"partners":["CENPH","CENPM","CENPK","CENPC","MAD1L1","MAD2L1","MIS18BP1"],"other_free_text":[]},"mechanistic_narrative":"CENP-I is a constitutive inner kinetochore protein built around N-terminal HEAT repeats that directly binds AT-rich centromeric DNA to stabilize CENP-A nucleosomes and promote new CENP-A loading, thereby maintaining centromere identity across cell divisions [PMID:36888657, PMID:10864871, PMID:35970865]. It occupies a central position in kinetochore hierarchy: CENP-I (together with CENP-H) is required for centromeric localization of CENP-C, and it recruits M18BP1 downstream of CENP-C to couple kinetochore structure to the CENP-A assembly pathway [PMID:11970896, PMID:26527398]. CENP-I is also essential for spindle assembly checkpoint signaling, generating stable kinetochore association of the RZZ complex and Mad1/Mad2 while inhibiting their dynein-mediated stripping; together with Aurora B kinase activity, CENP-I forms a molecular switch that sustains checkpoint signaling at unattached kinetochores until stable microtubule attachment is achieved [PMID:12640463, PMID:24862574]. Beyond mitosis, CENP-I promotes homologous-recombination-mediated DNA double-strand break repair by suppressing R-loop accumulation [PMID:34206916]."},"prefetch_data":{"uniprot":{"accession":"Q92674","full_name":"Centromere protein I","aliases":["FSH primary response protein 1","Follicle-stimulating hormone primary response protein","Interphase centromere complex protein 19","Leucine-rich primary response protein 1"],"length_aa":756,"mass_kda":86.7,"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. Required for the localization of CENPF, MAD1L1 and MAD2 (MAD2L1 or MAD2L2) to kinetochores. Involved in the response of gonadal tissues to follicle-stimulating hormone","subcellular_location":"Nucleus; Chromosome, centromere","url":"https://www.uniprot.org/uniprotkb/Q92674/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/CENPI","classification":"Common Essential","n_dependent_lines":852,"n_total_lines":1208,"dependency_fraction":0.7052980132450332},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[],"url":"https://opencell.sf.czbiohub.org/search/CENPI","total_profiled":1310},"omim":[{"mim_id":"611511","title":"MLF1-INTERACTING PROTEIN; MLF1IP","url":"https://www.omim.org/entry/611511"},{"mim_id":"611506","title":"CENTROMERIC PROTEIN Q; CENPQ","url":"https://www.omim.org/entry/611506"},{"mim_id":"611505","title":"CENTROMERIC PROTEIN P; CENPP","url":"https://www.omim.org/entry/611505"},{"mim_id":"611504","title":"CENTROMERIC PROTEIN O; CENPO","url":"https://www.omim.org/entry/611504"},{"mim_id":"611503","title":"CENTROMERIC PROTEIN L; CENPL","url":"https://www.omim.org/entry/611503"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Supported","locations":[{"location":"Nuclear bodies","reliability":"Supported"},{"location":"Nucleoplasm","reliability":"Additional"}],"tissue_specificity":"Tissue enhanced","tissue_distribution":"Detected in many","driving_tissues":[{"tissue":"bone marrow","ntpm":5.2},{"tissue":"lymphoid tissue","ntpm":4.9}],"url":"https://www.proteinatlas.org/search/CENPI"},"hgnc":{"alias_symbol":["LRPR1","CENP-I","Mis6"],"prev_symbol":["FSHPRH1"]},"alphafold":{"accession":"Q92674","domains":[{"cath_id":"-","chopping":"687-737","consensus_level":"medium","plddt":82.6702,"start":687,"end":737},{"cath_id":"1.25.40","chopping":"85-248","consensus_level":"medium","plddt":83.1731,"start":85,"end":248}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q92674","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q92674-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q92674-F1-predicted_aligned_error_v6.png","plddt_mean":73.75},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=CENPI","jax_strain_url":"https://www.jax.org/strain/search?query=CENPI"},"sequence":{"accession":"Q92674","fasta_url":"https://rest.uniprot.org/uniprotkb/Q92674.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q92674/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q92674"}},"corpus_meta":[{"pmid":"10864871","id":"PMC_10864871","title":"Requirement of Mis6 centromere connector for localizing a CENP-A-like protein in fission yeast.","date":"2000","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/10864871","citation_count":341,"is_preprint":false},{"pmid":"9230309","id":"PMC_9230309","title":"Mis6, a fission yeast inner centromere protein, acts during G1/S and forms specialized chromatin required for equal segregation.","date":"1997","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/9230309","citation_count":198,"is_preprint":false},{"pmid":"10398680","id":"PMC_10398680","title":"Proper metaphase spindle length is determined by centromere proteins Mis12 and Mis6 required for faithful chromosome segregation.","date":"1999","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/10398680","citation_count":178,"is_preprint":false},{"pmid":"12640463","id":"PMC_12640463","title":"Human CENP-I specifies localization of CENP-F, MAD1 and MAD2 to kinetochores and is essential for mitosis.","date":"2003","source":"Nature cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/12640463","citation_count":129,"is_preprint":false},{"pmid":"11970896","id":"PMC_11970896","title":"CENP-I is essential for centromere function in vertebrate cells.","date":"2002","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/11970896","citation_count":118,"is_preprint":false},{"pmid":"11782448","id":"PMC_11782448","title":"Ctf3p, the Mis6 budding yeast homolog, interacts with Mcm22p and Mcm16p at the yeast outer kinetochore.","date":"2002","source":"Genes & development","url":"https://pubmed.ncbi.nlm.nih.gov/11782448","citation_count":107,"is_preprint":false},{"pmid":"26527398","id":"PMC_26527398","title":"CENP-C and CENP-I are key connecting factors for kinetochore and CENP-A assembly.","date":"2015","source":"Journal of cell science","url":"https://pubmed.ncbi.nlm.nih.gov/26527398","citation_count":54,"is_preprint":false},{"pmid":"24862574","id":"PMC_24862574","title":"CENP-I and Aurora B act as a molecular switch that ties RZZ/Mad1 recruitment to kinetochore attachment status.","date":"2014","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/24862574","citation_count":42,"is_preprint":false},{"pmid":"24789708","id":"PMC_24789708","title":"Eic1 links Mis18 with the CCAN/Mis6/Ctf19 complex to promote CENP-A assembly.","date":"2014","source":"Open biology","url":"https://pubmed.ncbi.nlm.nih.gov/24789708","citation_count":37,"is_preprint":false},{"pmid":"15930132","id":"PMC_15930132","title":"Spindle checkpoint signaling requires the mis6 kinetochore subcomplex, which interacts with mad2 and mitotic spindles.","date":"2005","source":"Molecular biology of the cell","url":"https://pubmed.ncbi.nlm.nih.gov/15930132","citation_count":31,"is_preprint":false},{"pmid":"29936263","id":"PMC_29936263","title":"CENPI is overexpressed in colorectal cancer and regulates cell migration and invasion.","date":"2018","source":"Gene","url":"https://pubmed.ncbi.nlm.nih.gov/29936263","citation_count":24,"is_preprint":false},{"pmid":"21445296","id":"PMC_21445296","title":"Mis17 is a regulatory module of the Mis6-Mal2-Sim4 centromere complex that is required for the recruitment of CenH3/CENP-A in fission yeast.","date":"2011","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/21445296","citation_count":21,"is_preprint":false},{"pmid":"25375240","id":"PMC_25375240","title":"The kinetochore protein Kis1/Eic1/Mis19 ensures the integrity of mitotic spindles through maintenance of kinetochore factors Mis6/CENP-I and CENP-A.","date":"2014","source":"PloS one","url":"https://pubmed.ncbi.nlm.nih.gov/25375240","citation_count":16,"is_preprint":false},{"pmid":"21888900","id":"PMC_21888900","title":"Anti-CENPI autoantibodies in scleroderma patients with features of autoimmune liver diseases.","date":"2011","source":"Clinica chimica acta; international journal of clinical chemistry","url":"https://pubmed.ncbi.nlm.nih.gov/21888900","citation_count":16,"is_preprint":false},{"pmid":"8921378","id":"PMC_8921378","title":"Sequence and chromosomal location of a human homologue of LRPR1, an FSH primary response gene.","date":"1996","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/8921378","citation_count":15,"is_preprint":false},{"pmid":"36888657","id":"PMC_36888657","title":"CENP-I directly targets centromeric DNA to support CENP-A deposition and centromere maintenance.","date":"2023","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/36888657","citation_count":9,"is_preprint":false},{"pmid":"31912435","id":"PMC_31912435","title":"Centromere protein I (CENPI) is a candidate gene for X-linked steroid sensitive nephrotic syndrome.","date":"2020","source":"Journal of nephrology","url":"https://pubmed.ncbi.nlm.nih.gov/31912435","citation_count":8,"is_preprint":false},{"pmid":"37465344","id":"PMC_37465344","title":"Pan-Cancer Analysis Reveals CENPI as a Potential Biomarker and Therapeutic Target in Adrenocortical Carcinoma.","date":"2023","source":"Journal of inflammation research","url":"https://pubmed.ncbi.nlm.nih.gov/37465344","citation_count":7,"is_preprint":false},{"pmid":"34081091","id":"PMC_34081091","title":"Ctf3/CENP-I provides a docking site for the desumoylase Ulp2 at the kinetochore.","date":"2021","source":"The Journal of cell biology","url":"https://pubmed.ncbi.nlm.nih.gov/34081091","citation_count":6,"is_preprint":false},{"pmid":"34206916","id":"PMC_34206916","title":"Loss of CENP-I Impairs Homologous Recombination and Sensitizes Cells to PARP1 Inhibition.","date":"2021","source":"Cancers","url":"https://pubmed.ncbi.nlm.nih.gov/34206916","citation_count":5,"is_preprint":false},{"pmid":"40406242","id":"PMC_40406242","title":"The m6A reader YTHDF3 promotes TNBC progression by regulating CENPI stabilization.","date":"2025","source":"Frontiers in oncology","url":"https://pubmed.ncbi.nlm.nih.gov/40406242","citation_count":4,"is_preprint":false},{"pmid":"9701787","id":"PMC_9701787","title":"Differential regulation of leucine-rich primary response gene 1 (LRPR1) mRNA expression in rat testis and ovary.","date":"1998","source":"Molecular human reproduction","url":"https://pubmed.ncbi.nlm.nih.gov/9701787","citation_count":4,"is_preprint":false},{"pmid":"35970865","id":"PMC_35970865","title":"The Mis6 inner kinetochore subcomplex maintains CENP-A nucleosomes against centromeric non-coding transcription during mitosis.","date":"2022","source":"Communications biology","url":"https://pubmed.ncbi.nlm.nih.gov/35970865","citation_count":3,"is_preprint":false},{"pmid":"30967422","id":"PMC_30967422","title":"Negative Regulation of the Mis17-Mis6 Centromere Complex by mRNA Decay Pathway and EKC/KEOPS Complex in Schizosaccharomyces pombe.","date":"2019","source":"G3 (Bethesda, Md.)","url":"https://pubmed.ncbi.nlm.nih.gov/30967422","citation_count":2,"is_preprint":false},{"pmid":"32017295","id":"PMC_32017295","title":"Structural insights into the intramolecular interactions of centromere protein CENP-I.","date":"2020","source":"Journal of molecular recognition : JMR","url":"https://pubmed.ncbi.nlm.nih.gov/32017295","citation_count":2,"is_preprint":false},{"pmid":"40784804","id":"PMC_40784804","title":"[CENPI promotes the migration of liver cancer cells and the epithelial-mesenchymal transition process by activating the RAS/MEK/ERK signaling axis].","date":"2025","source":"Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology","url":"https://pubmed.ncbi.nlm.nih.gov/40784804","citation_count":1,"is_preprint":false},{"pmid":"41206587","id":"PMC_41206587","title":"FOXM1/CENPI axis regulation of proline and arginine metabolism in glioblastoma cells.","date":"2026","source":"Journal of neuropathology and experimental neurology","url":"https://pubmed.ncbi.nlm.nih.gov/41206587","citation_count":0,"is_preprint":false}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":13632,"output_tokens":2977,"usd":0.042776},"stage2":{"model":"claude-opus-4-6","input_tokens":6322,"output_tokens":2892,"usd":0.155865},"total_usd":0.198641,"stage1_batch_id":"msgbatch_011kfMfE81o6CTcyhVUqumuW","stage2_batch_id":"msgbatch_01DTputS6WNkH745x1nVSoBo","note":"batch pricing = 50% of standard"},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2003,\n      \"finding\": \"Human CENP-I is required for the localization of CENP-F and the spindle checkpoint proteins MAD1 and MAD2 to kinetochores; depletion of CENP-I causes G2 delay and inability to arrest in mitosis despite unattached kinetochores, with residual mitotic delay being MAD2-dependent and requiring collective signal from many unattached kinetochores.\",\n      \"method\": \"RNAi depletion, immunofluorescence, checkpoint analysis in human cells\",\n      \"journal\": \"Nature cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — clean depletion with defined cellular phenotypes, multiple orthogonal readouts, replicated concept across studies\",\n      \"pmids\": [\"12640463\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"CENP-I is a constitutive centromere component that colocalizes with CENP-A, -C, and -H throughout the cell cycle; loss of CENP-I in chicken DT40 cells causes prometaphase arrest with misaligned chromosomes; both CENP-I and CENP-H are required for localization of CENP-C but not CENP-A to the centromere.\",\n      \"method\": \"Conditional knockout in chicken DT40 cells, immunocytochemistry, co-localization analysis\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — conditional loss-of-function with defined molecular hierarchy, replicated in vertebrate model\",\n      \"pmids\": [\"11970896\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"CENP-I is required to generate stable association of the RZZ complex and Mad1 with kinetochores, and also inhibits their dynein-mediated stripping; Aurora B regulates RZZ/Mad1 association while CENP-I inhibits dissociation, together forming a molecular switch maintaining spindle checkpoint signal at prometaphase kinetochores until mature microtubule attachment.\",\n      \"method\": \"CENP-I depletion (RNAi), Aurora B inhibition, immunofluorescence, epistasis analysis in human cells\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — epistasis between CENP-I and Aurora B established with multiple orthogonal methods, clear mechanistic model\",\n      \"pmids\": [\"24862574\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2015,\n      \"finding\": \"CENP-I can recruit M18BP1 (Mis18BP1) and enhance its assembly at centromeres downstream of CENP-C, functioning as a key connecting factor between kinetochore structure and CENP-A assembly; tethering CENP-I to ectopic sites induced de novo CENP-A assembly by recruiting CENP-C and subsequently M18BP1.\",\n      \"method\": \"Tetracycline repressor tethering to synthetic alphoid DNA arrays in human artificial chromosomes, immunofluorescence\",\n      \"journal\": \"Journal of cell science\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — direct tethering experiment with functional readout, single study\",\n      \"pmids\": [\"26527398\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2023,\n      \"finding\": \"CENP-I directly interacts with centromeric DNA, preferentially recognizing AT-rich elements via a consecutive DNA-binding surface formed by conserved charged residues at the end of N-terminal HEAT repeats; DNA-binding-deficient mutants retain interaction with CENP-H/K and CENP-M but show significantly diminished centromeric localization, impaired chromosome alignment, and reduced centromeric loading of newly synthesized CENP-A; CENP-I stabilizes CENP-A nucleosomes by binding nucleosomal DNA rather than histones.\",\n      \"method\": \"In vitro DNA-binding assays, mutagenesis, co-immunoprecipitation, immunofluorescence, CENP-A loading assays in human cells\",\n      \"journal\": \"Proceedings of the National Academy of Sciences of the United States of America\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — in vitro reconstitution with mutagenesis, structural surface mapping, and functional cellular validation\",\n      \"pmids\": [\"36888657\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2020,\n      \"finding\": \"A conserved helix (α11) in CENP-I forms intramolecular interactions with N-terminal HEAT repeats; deletion of this helix in human CENP-I dramatically reduces interaction with CENP-H and CENP-M, and mutations of conserved residues on the helix specifically weaken binding to CENP-M but not CENP-H in HeLa cells.\",\n      \"method\": \"Structural analysis, in vitro aggregation assays, co-immunoprecipitation in HeLa cells, mutagenesis\",\n      \"journal\": \"Journal of molecular recognition\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 1-2 — mutagenesis with in vitro and cellular binding assays, single study\",\n      \"pmids\": [\"32017295\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"A conserved surface of Ctf3/CENP-I (budding yeast ortholog) provides a direct binding site for the desumoylase Ulp2; Ctf3 mutations disabling Ulp2 recruitment cause elevated inner kinetochore sumoylation and defective chromosome segregation.\",\n      \"method\": \"Structural analysis, mutagenesis, yeast genetics, sumoylation assays, chromosome segregation assays\",\n      \"journal\": \"The Journal of cell biology\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1-2 — structural surface identified, mutagenesis with functional readout, ortholog of CENP-I\",\n      \"pmids\": [\"34081091\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2021,\n      \"finding\": \"Loss of CENP-I impairs homologous recombination (HR) DSB repair while having no effect on non-homologous end-joining; CENP-I deficiency increases R-loop formation, and RNaseH1 expression restores HR capacity in CENP-I-deficient cells, linking CENP-I to R-loop resolution as part of its HR function.\",\n      \"method\": \"CENP-I knockout/knockdown, HR and NHEJ reporter assays, 53BP1 foci analysis, RNaseH1 rescue experiments, PARP inhibitor sensitivity assay\",\n      \"journal\": \"Cancers\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal assays establishing HR role, single study\",\n      \"pmids\": [\"34206916\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2022,\n      \"finding\": \"The inner kinetochore protein Mis6 (CENP-I ortholog in fission yeast) and Mis15 (CENP-N) retain pre-existing CENP-A at centromeres during mitosis by blocking non-coding RNA transcription at the central core centromere region; eliminating Mis6 during mitosis caused immediate loss of CENP-A due to upregulation of non-coding RNAs, and RNA polymerase II inhibition prevented CENP-A loss in mis6 mutants.\",\n      \"method\": \"Conditional depletion in fission yeast, immunofluorescence, RNA analysis, RNA Pol II inhibition rescue experiment\",\n      \"journal\": \"Communications biology\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — genetic epistasis with pharmacological rescue, fission yeast ortholog, single study\",\n      \"pmids\": [\"35970865\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2002,\n      \"finding\": \"Budding yeast Ctf3p (CENP-I ortholog) interacts with Mcm22p and Mcm16p at the outer kinetochore, and all three bind centromere DNA in a Ctf19p-dependent manner as shown by chromatin immunoprecipitation; unlike fission yeast Mis6, Ctf3p is not required for loading of the CENP-A homolog Cse4p, but Ctf3p and Ctf19p fail to bind centromere DNA in a cse4-1 mutant.\",\n      \"method\": \"Synthetic dosage lethality screen, two-hybrid interaction, chromatin immunoprecipitation, genetic epistasis in budding yeast\",\n      \"journal\": \"Genes & development\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — ChIP plus genetic epistasis, multiple orthogonal methods, foundational study\",\n      \"pmids\": [\"11782448\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2000,\n      \"finding\": \"The fission yeast CENP-A homolog (SpCENP-A) binding to inner centromeres depends on Mis6 (CENP-I ortholog); Mis6 acts during G1-S phase and is required for recruiting SpCENP-A to form proper centromere chromatin for chromosome segregation.\",\n      \"method\": \"Temperature-sensitive mutant analysis, genetic epistasis, cell cycle staging, chromosome segregation assays in fission yeast\",\n      \"journal\": \"Science\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — epistasis with cell cycle resolution, highly cited foundational study\",\n      \"pmids\": [\"10864871\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2005,\n      \"finding\": \"The Mis6-complex (containing fission yeast CENP-I ortholog) physically interacts with Mad2 under conditions where the Mad2-dependent checkpoint is activated, and is required for accumulation of Mad2 onto unattached kinetochores; the Mis6-complex also collaborates with the Nuf2-complex to monitor spindle-kinetochore attachment state.\",\n      \"method\": \"Co-immunoprecipitation, immunofluorescence, genetic analysis of kinetochore components in fission yeast\",\n      \"journal\": \"Molecular biology of the cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2-3 — direct interaction by co-IP plus genetic requirement, fission yeast ortholog, single study\",\n      \"pmids\": [\"15930132\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"CENP-I is a constitutive inner kinetochore protein that directly binds AT-rich centromeric DNA via its N-terminal HEAT repeats to stabilize CENP-A nucleosomes and promote new CENP-A loading, recruits M18BP1 downstream of CENP-C to maintain centromere identity, is required for localization of CENP-C, CENP-F, MAD1, and MAD2 to kinetochores, acts together with Aurora B as a molecular switch to sustain spindle checkpoint signaling by stabilizing RZZ/Mad1 at unattached kinetochores and inhibiting dynein-mediated stripping, plays a role in homologous recombination DSB repair through R-loop suppression, and in yeast its ortholog docks the desumoylase Ulp2 to regulate kinetochore sumoylation.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"CENP-I is a constitutive inner kinetochore protein built around N-terminal HEAT repeats that directly binds AT-rich centromeric DNA to stabilize CENP-A nucleosomes and promote new CENP-A loading, thereby maintaining centromere identity across cell divisions [PMID:36888657, PMID:10864871, PMID:35970865]. It occupies a central position in kinetochore hierarchy: CENP-I (together with CENP-H) is required for centromeric localization of CENP-C, and it recruits M18BP1 downstream of CENP-C to couple kinetochore structure to the CENP-A assembly pathway [PMID:11970896, PMID:26527398]. CENP-I is also essential for spindle assembly checkpoint signaling, generating stable kinetochore association of the RZZ complex and Mad1/Mad2 while inhibiting their dynein-mediated stripping; together with Aurora B kinase activity, CENP-I forms a molecular switch that sustains checkpoint signaling at unattached kinetochores until stable microtubule attachment is achieved [PMID:12640463, PMID:24862574]. Beyond mitosis, CENP-I promotes homologous-recombination-mediated DNA double-strand break repair by suppressing R-loop accumulation [PMID:34206916].\",\n  \"teleology\": [\n    {\n      \"year\": 2000,\n      \"claim\": \"The first functional link between a CENP-I ortholog and centromere chromatin was established when fission yeast Mis6 was shown to be required during G1-S for loading the CENP-A homolog, defining CENP-I as upstream of centromeric nucleosome specification.\",\n      \"evidence\": \"Temperature-sensitive mis6 mutant analysis with cell-cycle staging and chromosome segregation assays in S. pombe\",\n      \"pmids\": [\"10864871\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Mechanism by which Mis6 promotes CENP-A incorporation was unknown\", \"Whether the requirement was direct or indirect was unresolved\", \"Applicability to vertebrate centromeres not yet tested\"]\n    },\n    {\n      \"year\": 2002,\n      \"claim\": \"Parallel work in budding yeast (Ctf3p) and chicken DT40 cells placed CENP-I as a constitutive centromere component required for CENP-C localization and proper chromosome segregation, revealing a conserved kinetochore assembly hierarchy in which CENP-I and CENP-H act upstream of CENP-C but downstream of CENP-A.\",\n      \"evidence\": \"Conditional knockout in DT40 cells with co-localization analysis; ChIP and genetic epistasis in budding yeast\",\n      \"pmids\": [\"11970896\", \"11782448\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CENP-I contacts centromeric DNA directly was unknown\", \"Relationship between CENP-I and outer kinetochore assembly undefined in vertebrates\"]\n    },\n    {\n      \"year\": 2003,\n      \"claim\": \"Human CENP-I was shown to be required for kinetochore recruitment of CENP-F, MAD1, and MAD2, establishing its role as a platform for spindle assembly checkpoint signaling in human cells.\",\n      \"evidence\": \"RNAi depletion of CENP-I in human cells with immunofluorescence and checkpoint analysis\",\n      \"pmids\": [\"12640463\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether CENP-I directly interacts with checkpoint proteins or acts indirectly was unclear\", \"Mechanism by which residual MAD2-dependent delay persists after CENP-I depletion not resolved\"]\n    },\n    {\n      \"year\": 2005,\n      \"claim\": \"Physical interaction between the Mis6 complex (CENP-I ortholog) and Mad2 was demonstrated in fission yeast, providing evidence that the inner kinetochore directly engages checkpoint machinery rather than simply serving as a structural scaffold.\",\n      \"evidence\": \"Co-immunoprecipitation and genetic analysis in S. pombe under checkpoint-activating conditions\",\n      \"pmids\": [\"15930132\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether the interaction is direct or bridged by other kinetochore components was not determined\", \"Co-IP performed under one condition; generality across checkpoint states untested\"]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"The checkpoint role of CENP-I was refined to a dual function: generating stable RZZ/Mad1 association at kinetochores and inhibiting dynein-mediated stripping, working together with Aurora B as a molecular switch that sustains checkpoint signaling until mature microtubule attachment.\",\n      \"evidence\": \"Epistasis between CENP-I depletion and Aurora B inhibition in human cells with quantitative kinetochore localization analysis\",\n      \"pmids\": [\"24862574\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Direct biochemical basis for CENP-I inhibition of dynein-mediated stripping was not identified\", \"How attachment status feeds back to relieve CENP-I's inhibitory role remained unknown\"]\n    },\n    {\n      \"year\": 2015,\n      \"claim\": \"CENP-I was linked to centromere maintenance by its ability to recruit M18BP1 and induce de novo CENP-A assembly downstream of CENP-C, revealing it as a coupling factor between kinetochore structure and epigenetic centromere propagation.\",\n      \"evidence\": \"Ectopic tethering of CENP-I to synthetic alphoid arrays on human artificial chromosomes with immunofluorescence readout\",\n      \"pmids\": [\"26527398\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Physiological relevance of ectopic tethering system to endogenous centromeres not fully validated\", \"Whether CENP-I directly contacts M18BP1 or acts through intermediate factors was not resolved\"]\n    },\n    {\n      \"year\": 2020,\n      \"claim\": \"Structural dissection of a conserved intramolecular helix (α11) in CENP-I revealed that it mediates assembly with CENP-H and CENP-M through distinct binding interfaces, beginning to define the structural basis of the CENP-H/I/K/M sub-complex.\",\n      \"evidence\": \"Mutagenesis, in vitro aggregation assays, and co-immunoprecipitation in HeLa cells\",\n      \"pmids\": [\"32017295\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"No high-resolution structure of the CENP-H/I/K/M complex was available\", \"In vivo consequences of helix mutations on kinetochore function not tested\"]\n    },\n    {\n      \"year\": 2021,\n      \"claim\": \"Two distinct non-kinetochore functions were uncovered: in budding yeast, a conserved surface of Ctf3/CENP-I docks the desumoylase Ulp2 to regulate inner kinetochore sumoylation; in human cells, CENP-I promotes homologous recombination DSB repair by suppressing R-loop accumulation.\",\n      \"evidence\": \"Structural analysis and mutagenesis with sumoylation/segregation assays in yeast; HR/NHEJ reporter assays and RNaseH1 rescue in human cells\",\n      \"pmids\": [\"34081091\", \"34206916\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"Whether the Ulp2-docking function is conserved in human CENP-I is unknown\", \"Mechanism by which CENP-I suppresses R-loops (direct RNA binding vs indirect) was not determined\", \"Whether the HR function operates at centromeric or genome-wide loci is unclear\"]\n    },\n    {\n      \"year\": 2022,\n      \"claim\": \"Fission yeast Mis6 (CENP-I) was shown to retain pre-existing CENP-A during mitosis by blocking non-coding RNA transcription at centromeres, providing a mechanistic explanation for CENP-A stabilization distinct from direct DNA binding.\",\n      \"evidence\": \"Conditional depletion of Mis6 in mitotic S. pombe cells with RNA analysis and RNA Pol II inhibition rescue\",\n      \"pmids\": [\"35970865\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\"Whether this transcription-blocking mechanism operates in human cells is untested\", \"Relationship between this RNA-based mechanism and direct DNA-binding stabilization of CENP-A is unresolved\"]\n    },\n    {\n      \"year\": 2023,\n      \"claim\": \"The long-standing question of whether CENP-I contacts centromeric DNA directly was resolved: CENP-I binds AT-rich DNA through conserved charged residues at the end of N-terminal HEAT repeats, and this interaction is required for centromeric localization, chromosome alignment, and CENP-A loading — independent of CENP-H/K/M binding.\",\n      \"evidence\": \"In vitro DNA-binding assays, charge-reversal mutagenesis, co-immunoprecipitation, and CENP-A loading assays in human cells\",\n      \"pmids\": [\"36888657\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\"No atomic-resolution structure of CENP-I bound to nucleosomal DNA exists\", \"Whether DNA-binding and transcription-blocking are the same or separable mechanisms is unclear\"]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"A high-resolution structure of CENP-I bound to centromeric nucleosomal DNA is lacking, and the molecular basis by which CENP-I inhibits dynein-mediated stripping of checkpoint proteins, the conservation of the Ulp2-docking function in vertebrates, and the mechanism linking CENP-I to R-loop suppression remain unresolved.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\"No cryo-EM or crystal structure of CENP-I on nucleosomal DNA\", \"Dynein-stripping inhibition mechanism not biochemically defined\", \"R-loop suppression mechanism (direct vs indirect) uncharacterized\"]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0003677\", \"supporting_discovery_ids\": [4, 9]},\n      {\"term_id\": \"GO:0098772\", \"supporting_discovery_ids\": [2, 6]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0, 1, 4, 9]},\n      {\"term_id\": \"GO:0005634\", \"supporting_discovery_ids\": [7]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1, 2, 10]},\n      {\"term_id\": \"R-HSA-73894\", \"supporting_discovery_ids\": [7]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [4, 8, 10]}\n    ],\n    \"complexes\": [\n      \"CENP-H/I/K/M complex\",\n      \"Constitutive centromere-associated network (CCAN)\"\n    ],\n    \"partners\": [\n      \"CENPH\",\n      \"CENPM\",\n      \"CENPK\",\n      \"CENPC\",\n      \"MAD1L1\",\n      \"MAD2L1\",\n      \"MIS18BP1\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}