{"gene":"MIS18A","run_date":"2026-04-28T18:30:28","timeline":{"discoveries":[{"year":2007,"finding":"hMis18α (MIS18A), hMis18β, and M18BP1 form a complex that accumulates specifically at the telophase-G1 centromere and is essential for the subsequent recruitment of de novo-synthesized CENP-A. RNAi knockdown of any of the three subunits abolishes centromeric CENP-A recruitment, leading to misaligned chromosomes, anaphase missegregation, and interphase micronuclei. The histone deacetylase inhibitor trichostatin A suppresses loss of CENP-A recruitment in hMis18α RNAi cells, implicating acetylation status of centromere chromatin in the priming mechanism.","method":"RNAi knockdown, live-cell imaging, immunofluorescence, co-immunoprecipitation, cell-cycle staging","journal":"Developmental cell","confidence":"High","confidence_rationale":"Tier 2 — reciprocal co-IP, clean RNAi with defined cellular phenotypes, replicated across three subunits with pharmacological rescue","pmids":["17199038"],"is_preprint":false},{"year":2014,"finding":"Polo-like kinase 1 (Plk1) localizes to centromeres and is required to initiate CENP-A deposition by promoting the centromeric localization of the Mis18 complex, including MIS18A. CDK activity independently inhibits Mis18 complex assembly. Bypassing both regulatory steps uncouples CENP-A deposition from cell-cycle progression and causes mitotic defects, establishing a two-step Plk1/CDK regulatory paradigm for CENP-A deposition.","method":"Chemical-genetic Plk1 inhibition, CDK inhibitor treatment, RNAi, immunofluorescence, co-immunoprecipitation, CENP-A incorporation assay","journal":"Cell","confidence":"High","confidence_rationale":"Tier 2 — multiple orthogonal chemical and genetic perturbations with defined centromere phenotypes, strong mechanistic follow-up","pmids":["25036634"],"is_preprint":false},{"year":2016,"finding":"MIS18A and MIS18β form a heterotetramer through their C-terminal coiled-coil domains; this heterotetramer is required for M18BP1 binding and centromere recognition. HJURP is recruited to centromeres through a direct interaction between its centromere targeting domain and the MIS18A-MIS18β C-terminal coiled-coil domains. HJURP binding disrupts the MIS18A-MIS18β heterotetramer and removes MIS18A from centromeres, restricting CENP-A deposition to a single event per cell cycle.","method":"Co-immunoprecipitation, in vitro pulldown, size-exclusion chromatography, domain mapping, mutagenesis, immunofluorescence","journal":"Molecular cell","confidence":"High","confidence_rationale":"Tier 1–2 — direct biochemical reconstitution of heterotetramer, mutagenesis of coiled-coil domains, multiple orthogonal methods","pmids":["26942680"],"is_preprint":false},{"year":2016,"finding":"The acetyltransferase KAT7/HBO1/MYST2 interacts with the CENP-A assembly factor M18BP1 (a component of the Mis18 complex containing MIS18A). KAT7 knockout reduces centromeric CENP-A assembly and increases chromosome misalignment; tethering KAT7 to an ectopic alphoid DNA site removes H3K9me3 and stimulates CENP-A assembly, indicating that KAT7-containing acetyltransferases associated with the Mis18 complex provide competence for histone exchange on centromeric DNA.","method":"Co-immunoprecipitation, CRISPR knockout, immunofluorescence, chromatin tethering assay, ChIP","journal":"Developmental cell","confidence":"Medium","confidence_rationale":"Tier 2 — interaction with Mis18 complex shown by co-IP, functional KO phenotype; MIS18A itself not directly manipulated","pmids":["27270040"],"is_preprint":false},{"year":2017,"finding":"A human MIS18A:MIS18β complex forms a 4:2 hexamer (via Yippee domains) that binds two copies of M18BP1 through M18BP1's 140 N-terminal residues. CDK1 phosphorylation at two conserved sites in this M18BP1 N-terminal region destabilizes binding to the MIS18A:MIS18β scaffold, restricting Mis18 complex assembly and centromere recruitment to G1 phase.","method":"Structural biochemistry (SEC-MALS, cross-linking MS), recombinant protein reconstitution, phospho-mutagenesis, viral 2A co-expression system, immunofluorescence","journal":"eLife","confidence":"High","confidence_rationale":"Tier 1 — reconstituted hexamer in vitro, mutagenesis of CDK1 sites, multiple orthogonal structural and cell-based methods","pmids":["28059702"],"is_preprint":false},{"year":2025,"finding":"FAM72A, expressed from the divergent |Srgap2-Fam72a| master gene locus upon EGF stimulation, downregulates MIS18A (a tightly cell cycle-regulated gene) and interferes with AKT1 signaling by reducing phospho-AKT1 (Ser473), thereby favoring the MAPK1 route to promote cellular proliferation.","method":"Reporter assays (IGR promoter activity), siRNA knockdown, Western blotting, RT-qPCR, EGF stimulation paradigm","journal":"Cell cycle (Georgetown, Tex.)","confidence":"Low","confidence_rationale":"Tier 3 — single lab, single set of overexpression/knockdown experiments; upstream regulator identified but mechanism of MIS18A downregulation not directly elucidated","pmids":["41414706"],"is_preprint":false}],"current_model":"MIS18A forms a heterotetramer with MIS18β (via C-terminal coiled-coil/Yippee domains) that, together with M18BP1, assembles a hexameric Mis18 complex at telophase-G1 centromeres to prime centromeric chromatin (through acetylation regulation) for CENP-A deposition by HJURP; complex formation and centromere recruitment are cell-cycle gated by Plk1 (promoting) and CDK1 (inhibiting via M18BP1 phosphorylation), and HJURP binding subsequently disrupts the heterotetramer to restrict CENP-A loading to one event per cycle."},"narrative":{"teleology":[{"year":2007,"claim":"The discovery that MIS18A, MIS18β, and M18BP1 form a telophase-G1-specific centromere complex essential for de novo CENP-A loading established the first upstream licensing step for centromere maintenance, answering how cells prepare centromeric chromatin for histone variant deposition.","evidence":"RNAi knockdown of each subunit in human cells combined with live-cell imaging, co-IP, and pharmacological rescue with HDAC inhibitor trichostatin A","pmids":["17199038"],"confidence":"High","gaps":["Physical stoichiometry and architecture of the Mis18 complex were undefined","Direct enzymatic activity (e.g., acetyltransferase vs. recruitment of acetylases) was not resolved","How the complex is restricted to specific cell-cycle windows was unknown"]},{"year":2014,"claim":"Identification of Plk1 as a positive regulator and CDK activity as a negative regulator of Mis18 complex centromere recruitment answered how CENP-A deposition is coupled to cell-cycle progression, revealing a two-step kinase gating mechanism.","evidence":"Chemical-genetic Plk1 inhibition, CDK inhibitor treatment, RNAi, and CENP-A incorporation assays in human cells","pmids":["25036634"],"confidence":"High","gaps":["Direct phosphorylation sites on Mis18 subunits or M18BP1 were not mapped","How Plk1 mechanistically promotes centromere targeting of MIS18A was unclear"]},{"year":2016,"claim":"Biochemical reconstitution of the MIS18A–MIS18β heterotetramer and demonstration that HJURP binding disrupts it resolved the architectural basis of centromere priming and provided a self-limiting mechanism that restricts CENP-A loading to one event per cycle.","evidence":"In vitro pulldown, size-exclusion chromatography, coiled-coil domain mutagenesis, and immunofluorescence in human cells","pmids":["26942680"],"confidence":"High","gaps":["Atomic-resolution structure of the MIS18A–MIS18β heterotetramer was not available","Whether heterotetramer disruption is sufficient for centromere eviction in vivo was not directly tested"]},{"year":2016,"claim":"Linking the acetyltransferase KAT7 to the Mis18 complex via M18BP1 clarified the chromatin-modifying effector arm through which the complex primes centromeres for histone exchange, resolving a question raised by earlier HDAC inhibitor rescue experiments.","evidence":"Co-IP of KAT7 with M18BP1, CRISPR knockout, ChIP, and ectopic chromatin tethering assay","pmids":["27270040"],"confidence":"Medium","gaps":["MIS18A was not directly shown to contact KAT7; interaction is via M18BP1","Whether KAT7 is the sole acetyltransferase recruited by the Mis18 complex was not addressed"]},{"year":2017,"claim":"Reconstitution of a 4:2 MIS18A:MIS18β hexamer bound to two M18BP1 copies, and mapping of CDK1 phosphorylation sites on M18BP1 that block this interaction, provided a precise structural and regulatory model for cell-cycle-restricted Mis18 complex assembly.","evidence":"SEC-MALS, cross-linking mass spectrometry, recombinant reconstitution, phospho-mutagenesis, and immunofluorescence","pmids":["28059702"],"confidence":"High","gaps":["High-resolution structure (cryo-EM or crystal) of the full hexameric complex was not obtained","How Plk1 counteracts CDK1-mediated inhibition at the structural level remains unresolved"]},{"year":null,"claim":"Outstanding questions include the atomic-resolution structure of the complete Mis18 complex, the mechanism by which Plk1 promotes centromere targeting, and whether MIS18A plays roles beyond centromere priming (e.g., in response to proliferative signaling).","evidence":"","pmids":[],"confidence":"High","gaps":["No high-resolution structure of the full Mis18 hexamer or its centromere-bound state exists","The direct Plk1 substrate(s) within the Mis18 complex are not identified","Regulation of MIS18A expression by mitogenic signaling (e.g., FAM72A/EGF axis) is supported only by preliminary evidence"]}],"mechanism_profile":{"molecular_activity":[{"term_id":"GO:0060090","term_label":"molecular adaptor activity","supporting_discovery_ids":[0,2,4]}],"localization":[{"term_id":"GO:0005694","term_label":"chromosome","supporting_discovery_ids":[0,1,2,4]}],"pathway":[{"term_id":"R-HSA-1640170","term_label":"Cell Cycle","supporting_discovery_ids":[1,4]},{"term_id":"R-HSA-4839726","term_label":"Chromatin organization","supporting_discovery_ids":[0,3]}],"complexes":["Mis18 complex"],"partners":["MIS18BP1","OIP5","HJURP","PLK1","KAT7"],"other_free_text":[]},"mechanistic_narrative":"MIS18A is a core subunit of the Mis18 complex that primes centromeric chromatin for de novo CENP-A deposition during telophase and G1 phase. MIS18A heteroligomerizes with MIS18β through C-terminal coiled-coil and Yippee domains, forming a 4:2 hexameric scaffold that binds two copies of M18BP1 via its N-terminal region; this assembly is required for centromere recognition and subsequent recruitment of the CENP-A loading factor HJURP [PMID:17199038, PMID:26942680, PMID:28059702]. Cell-cycle gating of Mis18 complex formation is enforced by CDK1-mediated phosphorylation of M18BP1, which blocks its association with the MIS18A–MIS18β scaffold outside of G1, while Plk1 activity promotes centromeric localization of the complex [PMID:25036634, PMID:28059702]. HJURP binding to the MIS18A–MIS18β coiled-coil domains disrupts the heterotetramer and displaces MIS18A from centromeres, thereby limiting CENP-A deposition to a single event per cell cycle [PMID:26942680]."},"prefetch_data":{"uniprot":{"accession":"Q9NYP9","full_name":"Protein Mis18-alpha","aliases":["FAPP1-associated protein 1"],"length_aa":233,"mass_kda":25.9,"function":"Required for recruitment of CENPA to centromeres and normal chromosome segregation during mitosis","subcellular_location":"Nucleus; Chromosome; Chromosome, centromere","url":"https://www.uniprot.org/uniprotkb/Q9NYP9/entry"},"depmap":{"release":"DepMap","has_data":true,"is_common_essential":true,"resolved_as":"","url":"https://depmap.org/portal/gene/MIS18A","classification":"Common Essential","n_dependent_lines":1136,"n_total_lines":1208,"dependency_fraction":0.9403973509933775},"opencell":{"profiled":false,"resolved_as":"","ensg_id":"","cell_line_id":"","localizations":[],"interactors":[{"gene":"HDAC2","stoichiometry":0.2}],"url":"https://opencell.sf.czbiohub.org/search/MIS18A","total_profiled":1310},"omim":[{"mim_id":"618139","title":"MIS18-BINDING PROTEIN 1; MIS18BP1","url":"https://www.omim.org/entry/618139"},{"mim_id":"618137","title":"MIS18 KINETOCHORE PROTEIN A: MIS18A","url":"https://www.omim.org/entry/618137"},{"mim_id":"612667","title":"HOLLIDAY JUNCTION RECOGNITION PROTEIN; HJURP","url":"https://www.omim.org/entry/612667"},{"mim_id":"606020","title":"OPA-INTERACTING PROTEIN 5; OIP5","url":"https://www.omim.org/entry/606020"},{"mim_id":"604970","title":"AURORA KINASE B; AURKB","url":"https://www.omim.org/entry/604970"}],"hpa":{"profiled":true,"resolved_as":"","reliability":"Approved","locations":[{"location":"Nucleoplasm","reliability":"Approved"},{"location":"Cytosol","reliability":"Approved"}],"tissue_specificity":"Low tissue specificity","tissue_distribution":"Detected in all","driving_tissues":[],"url":"https://www.proteinatlas.org/search/MIS18A"},"hgnc":{"alias_symbol":["B28","FASP1","hMis18alpha"],"prev_symbol":["C21orf46","C21orf45"]},"alphafold":{"accession":"Q9NYP9","domains":[{"cath_id":"-","chopping":"97-172","consensus_level":"high","plddt":94.7524,"start":97,"end":172}],"viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NYP9","model_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NYP9-F1-model_v6.cif","pae_url":"https://alphafold.ebi.ac.uk/files/AF-Q9NYP9-F1-predicted_aligned_error_v6.png","plddt_mean":79.12},"mouse_models":{"mgi_url":"https://www.informatics.jax.org/marker/summary?nomen=MIS18A","jax_strain_url":"https://www.jax.org/strain/search?query=MIS18A"},"sequence":{"accession":"Q9NYP9","fasta_url":"https://rest.uniprot.org/uniprotkb/Q9NYP9.fasta","uniprot_url":"https://www.uniprot.org/uniprotkb/Q9NYP9/entry","alphafold_viewer_url":"https://alphafold.ebi.ac.uk/entry/Q9NYP9"}},"corpus_meta":[{"pmid":"15897552","id":"PMC_15897552","title":"Paclitaxel after doxorubicin plus cyclophosphamide as adjuvant chemotherapy for node-positive breast cancer: results from NSABP B-28.","date":"2005","source":"Journal of clinical oncology : official journal of the American Society of Clinical Oncology","url":"https://pubmed.ncbi.nlm.nih.gov/15897552","citation_count":466,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"9708987","id":"PMC_9708987","title":"Interactions of phenol and m-cresol in the insulin hexamer, and their effect on the association properties of B28 pro --> Asp insulin analogues.","date":"1998","source":"Biochemistry","url":"https://pubmed.ncbi.nlm.nih.gov/9708987","citation_count":102,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"17309503","id":"PMC_17309503","title":"Effectiveness of cell-adsorbed bacteriocin produced by Lactobacillus curvatus CWBI-B28 and selected essential oils to control Listeria monocytogenes in pork meat during cold storage.","date":"2007","source":"Letters in applied microbiology","url":"https://pubmed.ncbi.nlm.nih.gov/17309503","citation_count":18,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"16715806","id":"PMC_16715806","title":"Bacteriocin activity by Lactobacillus curvatus CWBI-B28 to inactivate Listeria monocytogenes in cold-smoked salmon during 4 degrees C storage.","date":"2006","source":"Journal of food protection","url":"https://pubmed.ncbi.nlm.nih.gov/16715806","citation_count":18,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"34441683","id":"PMC_34441683","title":"Functional Genomic Insights into Probiotic Bacillus siamensis Strain B28 from Traditional Korean Fermented Kimchi.","date":"2021","source":"Foods (Basel, Switzerland)","url":"https://pubmed.ncbi.nlm.nih.gov/34441683","citation_count":15,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"8483809","id":"PMC_8483809","title":"Preparation of N epsilon B28-monoazidobenzoyl insulin-like growth factor I and photoaffinity labeling of insulin-like growth factor I receptor.","date":"1993","source":"Peptides","url":"https://pubmed.ncbi.nlm.nih.gov/8483809","citation_count":4,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"31576812","id":"PMC_31576812","title":"LncRNAs down-regulate Myh1, Casr, and Mis18a expression in the Substantia Nigra of aged male rats.","date":"2019","source":"Aging","url":"https://pubmed.ncbi.nlm.nih.gov/31576812","citation_count":2,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"38910901","id":"PMC_38910901","title":"MIS18A upregulation promotes cell viability, migration and tumor immune evasion in lung adenocarcinoma.","date":"2024","source":"Oncology letters","url":"https://pubmed.ncbi.nlm.nih.gov/38910901","citation_count":1,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"41414706","id":"PMC_41414706","title":"Cell cycle-regulated expression of Fam72a from the |Srgap2-Fam72a| master gene leads to Mis18a downregulation.","date":"2025","source":"Cell cycle (Georgetown, Tex.)","url":"https://pubmed.ncbi.nlm.nih.gov/41414706","citation_count":0,"is_preprint":false,"source_track":"pubmed_title"},{"pmid":"12477932","id":"PMC_12477932","title":"Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences.","date":"2002","source":"Proceedings of the National Academy of Sciences of the United States of America","url":"https://pubmed.ncbi.nlm.nih.gov/12477932","citation_count":1479,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26186194","id":"PMC_26186194","title":"The BioPlex Network: A Systematic Exploration of the Human Interactome.","date":"2015","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26186194","citation_count":1118,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28514442","id":"PMC_28514442","title":"Architecture of the human interactome defines protein communities and disease networks.","date":"2017","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/28514442","citation_count":1085,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"25416956","id":"PMC_25416956","title":"A proteome-scale map of the human interactome network.","date":"2014","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/25416956","citation_count":977,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32296183","id":"PMC_32296183","title":"A reference map of the human binary protein interactome.","date":"2020","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/32296183","citation_count":849,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"19490893","id":"PMC_19490893","title":"A genome-wide RNAi screen identifies multiple synthetic lethal interactions with the Ras oncogene.","date":"2009","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/19490893","citation_count":843,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"10830953","id":"PMC_10830953","title":"The DNA sequence of human chromosome 21.","date":"2000","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/10830953","citation_count":808,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"33961781","id":"PMC_33961781","title":"Dual proteome-scale networks reveal cell-specific remodeling of the human interactome.","date":"2021","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/33961781","citation_count":705,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21873635","id":"PMC_21873635","title":"Phylogenetic-based propagation of functional annotations within the Gene Ontology consortium.","date":"2011","source":"Briefings in bioinformatics","url":"https://pubmed.ncbi.nlm.nih.gov/21873635","citation_count":656,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"11001876","id":"PMC_11001876","title":"Identification of pleckstrin-homology-domain-containing proteins with novel phosphoinositide-binding specificities.","date":"2000","source":"The Biochemical journal","url":"https://pubmed.ncbi.nlm.nih.gov/11001876","citation_count":480,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"15489334","id":"PMC_15489334","title":"The status, quality, and expansion of the NIH full-length cDNA project: the Mammalian Gene Collection (MGC).","date":"2004","source":"Genome research","url":"https://pubmed.ncbi.nlm.nih.gov/15489334","citation_count":438,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26638075","id":"PMC_26638075","title":"A Dynamic Protein Interaction Landscape of the Human Centrosome-Cilium Interface.","date":"2015","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/26638075","citation_count":433,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"35271311","id":"PMC_35271311","title":"OpenCell: Endogenous tagging for the cartography of human cellular organization.","date":"2022","source":"Science (New York, N.Y.)","url":"https://pubmed.ncbi.nlm.nih.gov/35271311","citation_count":432,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"17043677","id":"PMC_17043677","title":"Disrupted in Schizophrenia 1 Interactome: evidence for the close connectivity of risk genes and a potential synaptic basis for schizophrenia.","date":"2006","source":"Molecular psychiatry","url":"https://pubmed.ncbi.nlm.nih.gov/17043677","citation_count":345,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"17199038","id":"PMC_17199038","title":"Priming of centromere for CENP-A recruitment by human hMis18alpha, hMis18beta, and M18BP1.","date":"2007","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/17199038","citation_count":338,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"24270810","id":"PMC_24270810","title":"High-content genome-wide RNAi screens identify regulators of parkin upstream of mitophagy.","date":"2013","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/24270810","citation_count":297,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"17207965","id":"PMC_17207965","title":"hORFeome v3.1: a resource of human open reading frames representing over 10,000 human genes.","date":"2007","source":"Genomics","url":"https://pubmed.ncbi.nlm.nih.gov/17207965","citation_count":222,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21988832","id":"PMC_21988832","title":"Toward an understanding of the protein interaction network of the human liver.","date":"2011","source":"Molecular systems biology","url":"https://pubmed.ncbi.nlm.nih.gov/21988832","citation_count":207,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"29568061","id":"PMC_29568061","title":"An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping of protein interactions and subcellular localizations.","date":"2018","source":"Nature communications","url":"https://pubmed.ncbi.nlm.nih.gov/29568061","citation_count":201,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"21516116","id":"PMC_21516116","title":"Next-generation sequencing to generate interactome datasets.","date":"2011","source":"Nature methods","url":"https://pubmed.ncbi.nlm.nih.gov/21516116","citation_count":200,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"25036634","id":"PMC_25036634","title":"Polo-like kinase 1 licenses CENP-A deposition at centromeres.","date":"2014","source":"Cell","url":"https://pubmed.ncbi.nlm.nih.gov/25036634","citation_count":152,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32877691","id":"PMC_32877691","title":"A High-Density Human Mitochondrial Proximity Interaction Network.","date":"2020","source":"Cell metabolism","url":"https://pubmed.ncbi.nlm.nih.gov/32877691","citation_count":148,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"18029348","id":"PMC_18029348","title":"Toward a confocal subcellular atlas of the human proteome.","date":"2007","source":"Molecular & cellular proteomics : MCP","url":"https://pubmed.ncbi.nlm.nih.gov/18029348","citation_count":114,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32707033","id":"PMC_32707033","title":"Kinase Interaction Network Expands Functional and Disease Roles of Human Kinases.","date":"2020","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/32707033","citation_count":88,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26942680","id":"PMC_26942680","title":"Licensing of Centromeric Chromatin Assembly through the Mis18α-Mis18β Heterotetramer.","date":"2016","source":"Molecular cell","url":"https://pubmed.ncbi.nlm.nih.gov/26942680","citation_count":84,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"27270040","id":"PMC_27270040","title":"KAT7/HBO1/MYST2 Regulates CENP-A Chromatin Assembly by Antagonizing Suv39h1-Mediated Centromere Inactivation.","date":"2016","source":"Developmental cell","url":"https://pubmed.ncbi.nlm.nih.gov/27270040","citation_count":79,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"32814053","id":"PMC_32814053","title":"Interactome Mapping Provides a Network of Neurodegenerative Disease Proteins and Uncovers Widespread Protein Aggregation in Affected Brains.","date":"2020","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/32814053","citation_count":79,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"26972000","id":"PMC_26972000","title":"Substrate-Trapped Interactors of PHD3 and FIH Cluster in Distinct Signaling Pathways.","date":"2016","source":"Cell reports","url":"https://pubmed.ncbi.nlm.nih.gov/26972000","citation_count":77,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"28059702","id":"PMC_28059702","title":"CDK-regulated dimerization of M18BP1 on a Mis18 hexamer is necessary for CENP-A loading.","date":"2017","source":"eLife","url":"https://pubmed.ncbi.nlm.nih.gov/28059702","citation_count":72,"is_preprint":false,"source_track":"gene2pubmed"},{"pmid":"36424410","id":"PMC_36424410","title":"MYC multimers shield stalled replication forks from RNA polymerase.","date":"2022","source":"Nature","url":"https://pubmed.ncbi.nlm.nih.gov/36424410","citation_count":63,"is_preprint":false,"source_track":"gene2pubmed"}],"cost":{"stage1":{"model":"claude-sonnet-4-6","input_tokens":5756,"output_tokens":561,"usd":0.012842},"stage2":{"model":"claude-opus-4-6","input_tokens":3724,"output_tokens":870,"usd":0.060555},"total_usd":0.22323,"stage1_batch_id":"msgbatch_01Mgbec4vM6c9vXht7JVJzss","stage2_batch_id":"msgbatch_01JkuuH5RB2BdJNsgnhjbC6s","note":"batch pricing = 50% of standard","round2_stage1":{"model":"claude-sonnet-4-6","input_tokens":17334,"output_tokens":1859,"usd":0.039943},"round2_rules_fired":"R3","round2_stage2":{"model":"claude-opus-4-6","input_tokens":5377,"output_tokens":1855,"usd":0.10989}},"stage1_raw":"```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2025,\n      \"finding\": \"FAM72A downregulates MIS18A expression during the cell cycle; upon EGF stimulation, FAM72A expression is driven by the intergenic region between SRGAP2 and FAM72A acting as a divergent promoter, and FAM72A additionally interferes with the AKT1 signaling pathway by reducing phospho-AKT1 (Ser473), thereby favoring the MAPK1 route to promote cellular proliferation.\",\n      \"method\": \"Cell-based assay with EGF stimulation, RT-qPCR, western blot, and promoter activity analysis\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 3 — single lab, single paper with multiple readouts (RT-qPCR, western blot, promoter assay) but no in vitro reconstitution or genetic epistasis\",\n      \"pmids\": [\"41414706\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2024,\n      \"finding\": \"siRNA-mediated knockdown of MIS18A in lung cancer cells reduces proliferative and migratory capacities, implicating MIS18A in cell cycle regulation and immune-related pathway activity.\",\n      \"method\": \"siRNA knockdown in lung cancer cell lines with proliferation and migration assays\",\n      \"journal\": \"Oncology letters\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, single method (siRNA KD) with phenotypic readout but no pathway placement or molecular mechanism defined\",\n      \"pmids\": [\"38910901\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MIS18A is a tightly cell cycle-regulated kinetochore protein whose expression is downregulated by FAM72A (itself driven by EGF-stimulated divergent promoter activity of the SRGAP2-FAM72A intergenic region), and loss of MIS18A reduces cancer cell proliferation and migration, consistent with a role in cell cycle progression, though the precise molecular mechanism at the kinetochore remains to be fully defined in the available literature.\"\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"discoveries\": [\n    {\n      \"year\": 2007,\n      \"finding\": \"hMis18α (MIS18A), hMis18β, and M18BP1 form a complex that accumulates specifically at the telophase-G1 centromere and is essential for the subsequent recruitment of de novo-synthesized CENP-A. RNAi knockdown of any of the three subunits abolishes centromeric CENP-A recruitment, leading to misaligned chromosomes, anaphase missegregation, and interphase micronuclei. The histone deacetylase inhibitor trichostatin A suppresses loss of CENP-A recruitment in hMis18α RNAi cells, implicating acetylation status of centromere chromatin in the priming mechanism.\",\n      \"method\": \"RNAi knockdown, live-cell imaging, immunofluorescence, co-immunoprecipitation, cell-cycle staging\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — reciprocal co-IP, clean RNAi with defined cellular phenotypes, replicated across three subunits with pharmacological rescue\",\n      \"pmids\": [\"17199038\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2014,\n      \"finding\": \"Polo-like kinase 1 (Plk1) localizes to centromeres and is required to initiate CENP-A deposition by promoting the centromeric localization of the Mis18 complex, including MIS18A. CDK activity independently inhibits Mis18 complex assembly. Bypassing both regulatory steps uncouples CENP-A deposition from cell-cycle progression and causes mitotic defects, establishing a two-step Plk1/CDK regulatory paradigm for CENP-A deposition.\",\n      \"method\": \"Chemical-genetic Plk1 inhibition, CDK inhibitor treatment, RNAi, immunofluorescence, co-immunoprecipitation, CENP-A incorporation assay\",\n      \"journal\": \"Cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 2 — multiple orthogonal chemical and genetic perturbations with defined centromere phenotypes, strong mechanistic follow-up\",\n      \"pmids\": [\"25036634\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"MIS18A and MIS18β form a heterotetramer through their C-terminal coiled-coil domains; this heterotetramer is required for M18BP1 binding and centromere recognition. HJURP is recruited to centromeres through a direct interaction between its centromere targeting domain and the MIS18A-MIS18β C-terminal coiled-coil domains. HJURP binding disrupts the MIS18A-MIS18β heterotetramer and removes MIS18A from centromeres, restricting CENP-A deposition to a single event per cell cycle.\",\n      \"method\": \"Co-immunoprecipitation, in vitro pulldown, size-exclusion chromatography, domain mapping, mutagenesis, immunofluorescence\",\n      \"journal\": \"Molecular cell\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1–2 — direct biochemical reconstitution of heterotetramer, mutagenesis of coiled-coil domains, multiple orthogonal methods\",\n      \"pmids\": [\"26942680\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2016,\n      \"finding\": \"The acetyltransferase KAT7/HBO1/MYST2 interacts with the CENP-A assembly factor M18BP1 (a component of the Mis18 complex containing MIS18A). KAT7 knockout reduces centromeric CENP-A assembly and increases chromosome misalignment; tethering KAT7 to an ectopic alphoid DNA site removes H3K9me3 and stimulates CENP-A assembly, indicating that KAT7-containing acetyltransferases associated with the Mis18 complex provide competence for histone exchange on centromeric DNA.\",\n      \"method\": \"Co-immunoprecipitation, CRISPR knockout, immunofluorescence, chromatin tethering assay, ChIP\",\n      \"journal\": \"Developmental cell\",\n      \"confidence\": \"Medium\",\n      \"confidence_rationale\": \"Tier 2 — interaction with Mis18 complex shown by co-IP, functional KO phenotype; MIS18A itself not directly manipulated\",\n      \"pmids\": [\"27270040\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2017,\n      \"finding\": \"A human MIS18A:MIS18β complex forms a 4:2 hexamer (via Yippee domains) that binds two copies of M18BP1 through M18BP1's 140 N-terminal residues. CDK1 phosphorylation at two conserved sites in this M18BP1 N-terminal region destabilizes binding to the MIS18A:MIS18β scaffold, restricting Mis18 complex assembly and centromere recruitment to G1 phase.\",\n      \"method\": \"Structural biochemistry (SEC-MALS, cross-linking MS), recombinant protein reconstitution, phospho-mutagenesis, viral 2A co-expression system, immunofluorescence\",\n      \"journal\": \"eLife\",\n      \"confidence\": \"High\",\n      \"confidence_rationale\": \"Tier 1 — reconstituted hexamer in vitro, mutagenesis of CDK1 sites, multiple orthogonal structural and cell-based methods\",\n      \"pmids\": [\"28059702\"],\n      \"is_preprint\": false\n    },\n    {\n      \"year\": 2025,\n      \"finding\": \"FAM72A, expressed from the divergent |Srgap2-Fam72a| master gene locus upon EGF stimulation, downregulates MIS18A (a tightly cell cycle-regulated gene) and interferes with AKT1 signaling by reducing phospho-AKT1 (Ser473), thereby favoring the MAPK1 route to promote cellular proliferation.\",\n      \"method\": \"Reporter assays (IGR promoter activity), siRNA knockdown, Western blotting, RT-qPCR, EGF stimulation paradigm\",\n      \"journal\": \"Cell cycle (Georgetown, Tex.)\",\n      \"confidence\": \"Low\",\n      \"confidence_rationale\": \"Tier 3 — single lab, single set of overexpression/knockdown experiments; upstream regulator identified but mechanism of MIS18A downregulation not directly elucidated\",\n      \"pmids\": [\"41414706\"],\n      \"is_preprint\": false\n    }\n  ],\n  \"current_model\": \"MIS18A forms a heterotetramer with MIS18β (via C-terminal coiled-coil/Yippee domains) that, together with M18BP1, assembles a hexameric Mis18 complex at telophase-G1 centromeres to prime centromeric chromatin (through acetylation regulation) for CENP-A deposition by HJURP; complex formation and centromere recruitment are cell-cycle gated by Plk1 (promoting) and CDK1 (inhibiting via M18BP1 phosphorylation), and HJURP binding subsequently disrupts the heterotetramer to restrict CENP-A loading to one event per cycle.\"\n}\n```","stage2_raw":"```json\n{\n  \"mechanistic_narrative\": \"MIS18A is a cell cycle-regulated kinetochore-associated protein whose expression is negatively regulated by FAM72A; EGF stimulation drives FAM72A via a divergent promoter in the SRGAP2-FAM72A intergenic region, which in turn downregulates MIS18A and modulates signaling through reduced AKT1 phosphorylation (Ser473) while favoring the MAPK1 proliferation route [PMID:41414706]. siRNA-mediated depletion of MIS18A in lung cancer cells reduces both proliferation and migration, consistent with a functional role in cell cycle progression [PMID:38910901].\",\n  \"teleology\": [\n    {\n      \"year\": 2024,\n      \"claim\": \"Establishing that MIS18A loss-of-function impairs cancer cell proliferation and migration answered whether MIS18A is functionally required for these processes, but left the molecular mechanism undefined.\",\n      \"evidence\": \"siRNA knockdown of MIS18A in lung cancer cell lines with proliferation and migration assays\",\n      \"pmids\": [\"38910901\"],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"Single siRNA approach without rescue or orthogonal genetic validation\",\n        \"No molecular mechanism linking MIS18A to proliferation or migration pathways\",\n        \"No assessment of kinetochore integrity or chromosome segregation upon MIS18A depletion\"\n      ]\n    },\n    {\n      \"year\": 2025,\n      \"claim\": \"Identifying FAM72A as a negative regulator of MIS18A expression downstream of EGF stimulation placed MIS18A within a growth-factor-responsive transcriptional circuit and linked its regulation to AKT1/MAPK1 signaling balance.\",\n      \"evidence\": \"EGF stimulation in cell-based assays with RT-qPCR, western blot, and promoter activity analysis\",\n      \"pmids\": [\"41414706\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"Mechanism by which FAM72A downregulates MIS18A (transcriptional vs. post-transcriptional) is not resolved\",\n        \"Direct physical interaction between FAM72A and MIS18A or their promoters not demonstrated\",\n        \"Functional consequences of MIS18A downregulation by FAM72A on kinetochore assembly not tested\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"The precise molecular function of MIS18A at the kinetochore — including whether it acts as a structural scaffold, a recruitment factor for CENP-A loading, or has enzymatic activity — remains undefined in the available direct experimental literature.\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"Low\",\n      \"gaps\": [\n        \"No biochemical reconstitution of MIS18A activity\",\n        \"No structural data for MIS18A or its complexes\",\n        \"Relationship between MIS18A's kinetochore role and the proliferation/migration phenotypes not established\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [],\n    \"localization\": [],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [0, 1]},\n      {\"term_id\": \"R-HSA-162582\", \"supporting_discovery_ids\": [0]}\n    ],\n    \"complexes\": [],\n    \"partners\": [\"FAM72A\"],\n    \"other_free_text\": []\n  }\n}\n```\n\n--- ROUND 2 (R2/R3, augmented corpus) ---\n\n```json\n{\n  \"mechanistic_narrative\": \"MIS18A is a core subunit of the Mis18 complex that primes centromeric chromatin for de novo CENP-A deposition during telophase and G1 phase. MIS18A heteroligomerizes with MIS18β through C-terminal coiled-coil and Yippee domains, forming a 4:2 hexameric scaffold that binds two copies of M18BP1 via its N-terminal region; this assembly is required for centromere recognition and subsequent recruitment of the CENP-A loading factor HJURP [PMID:17199038, PMID:26942680, PMID:28059702]. Cell-cycle gating of Mis18 complex formation is enforced by CDK1-mediated phosphorylation of M18BP1, which blocks its association with the MIS18A–MIS18β scaffold outside of G1, while Plk1 activity promotes centromeric localization of the complex [PMID:25036634, PMID:28059702]. HJURP binding to the MIS18A–MIS18β coiled-coil domains disrupts the heterotetramer and displaces MIS18A from centromeres, thereby limiting CENP-A deposition to a single event per cell cycle [PMID:26942680].\",\n  \"teleology\": [\n    {\n      \"year\": 2007,\n      \"claim\": \"The discovery that MIS18A, MIS18β, and M18BP1 form a telophase-G1-specific centromere complex essential for de novo CENP-A loading established the first upstream licensing step for centromere maintenance, answering how cells prepare centromeric chromatin for histone variant deposition.\",\n      \"evidence\": \"RNAi knockdown of each subunit in human cells combined with live-cell imaging, co-IP, and pharmacological rescue with HDAC inhibitor trichostatin A\",\n      \"pmids\": [\"17199038\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Physical stoichiometry and architecture of the Mis18 complex were undefined\",\n        \"Direct enzymatic activity (e.g., acetyltransferase vs. recruitment of acetylases) was not resolved\",\n        \"How the complex is restricted to specific cell-cycle windows was unknown\"\n      ]\n    },\n    {\n      \"year\": 2014,\n      \"claim\": \"Identification of Plk1 as a positive regulator and CDK activity as a negative regulator of Mis18 complex centromere recruitment answered how CENP-A deposition is coupled to cell-cycle progression, revealing a two-step kinase gating mechanism.\",\n      \"evidence\": \"Chemical-genetic Plk1 inhibition, CDK inhibitor treatment, RNAi, and CENP-A incorporation assays in human cells\",\n      \"pmids\": [\"25036634\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Direct phosphorylation sites on Mis18 subunits or M18BP1 were not mapped\",\n        \"How Plk1 mechanistically promotes centromere targeting of MIS18A was unclear\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Biochemical reconstitution of the MIS18A–MIS18β heterotetramer and demonstration that HJURP binding disrupts it resolved the architectural basis of centromere priming and provided a self-limiting mechanism that restricts CENP-A loading to one event per cycle.\",\n      \"evidence\": \"In vitro pulldown, size-exclusion chromatography, coiled-coil domain mutagenesis, and immunofluorescence in human cells\",\n      \"pmids\": [\"26942680\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"Atomic-resolution structure of the MIS18A–MIS18β heterotetramer was not available\",\n        \"Whether heterotetramer disruption is sufficient for centromere eviction in vivo was not directly tested\"\n      ]\n    },\n    {\n      \"year\": 2016,\n      \"claim\": \"Linking the acetyltransferase KAT7 to the Mis18 complex via M18BP1 clarified the chromatin-modifying effector arm through which the complex primes centromeres for histone exchange, resolving a question raised by earlier HDAC inhibitor rescue experiments.\",\n      \"evidence\": \"Co-IP of KAT7 with M18BP1, CRISPR knockout, ChIP, and ectopic chromatin tethering assay\",\n      \"pmids\": [\"27270040\"],\n      \"confidence\": \"Medium\",\n      \"gaps\": [\n        \"MIS18A was not directly shown to contact KAT7; interaction is via M18BP1\",\n        \"Whether KAT7 is the sole acetyltransferase recruited by the Mis18 complex was not addressed\"\n      ]\n    },\n    {\n      \"year\": 2017,\n      \"claim\": \"Reconstitution of a 4:2 MIS18A:MIS18β hexamer bound to two M18BP1 copies, and mapping of CDK1 phosphorylation sites on M18BP1 that block this interaction, provided a precise structural and regulatory model for cell-cycle-restricted Mis18 complex assembly.\",\n      \"evidence\": \"SEC-MALS, cross-linking mass spectrometry, recombinant reconstitution, phospho-mutagenesis, and immunofluorescence\",\n      \"pmids\": [\"28059702\"],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"High-resolution structure (cryo-EM or crystal) of the full hexameric complex was not obtained\",\n        \"How Plk1 counteracts CDK1-mediated inhibition at the structural level remains unresolved\"\n      ]\n    },\n    {\n      \"year\": null,\n      \"claim\": \"Outstanding questions include the atomic-resolution structure of the complete Mis18 complex, the mechanism by which Plk1 promotes centromere targeting, and whether MIS18A plays roles beyond centromere priming (e.g., in response to proliferative signaling).\",\n      \"evidence\": \"\",\n      \"pmids\": [],\n      \"confidence\": \"High\",\n      \"gaps\": [\n        \"No high-resolution structure of the full Mis18 hexamer or its centromere-bound state exists\",\n        \"The direct Plk1 substrate(s) within the Mis18 complex are not identified\",\n        \"Regulation of MIS18A expression by mitogenic signaling (e.g., FAM72A/EGF axis) is supported only by preliminary evidence\"\n      ]\n    }\n  ],\n  \"mechanism_profile\": {\n    \"molecular_activity\": [\n      {\"term_id\": \"GO:0060090\", \"supporting_discovery_ids\": [0, 2, 4]}\n    ],\n    \"localization\": [\n      {\"term_id\": \"GO:0005694\", \"supporting_discovery_ids\": [0, 1, 2, 4]}\n    ],\n    \"pathway\": [\n      {\"term_id\": \"R-HSA-1640170\", \"supporting_discovery_ids\": [1, 4]},\n      {\"term_id\": \"R-HSA-4839726\", \"supporting_discovery_ids\": [0, 3]}\n    ],\n    \"complexes\": [\n      \"Mis18 complex\"\n    ],\n    \"partners\": [\n      \"MIS18BP1\",\n      \"OIP5\",\n      \"HJURP\",\n      \"PLK1\",\n      \"KAT7\"\n    ],\n    \"other_free_text\": []\n  }\n}\n```"}