title: A Dual Inhibitory Mechanism Sufficient to Maintain Cell-Cycle-Restricted CENP-A Assembly.
abstract: Chromatin featuring the H3 variant CENP-A at the centromere is critical for its mitotic function and epigenetic maintenance. Assembly of centromeric chromatin is restricted to G1 phase through inhibitory action of Cdk1/2 kinases in other phases of the cell cycle. Here, we identify the two key targets sufficient to maintain cell-cycle control of CENP-A assembly. We uncovered a single phosphorylation site in the licensing factor M18BP1 and a cyclin A binding site in the CENP-A chaperone, HJURP, that mediated specific inhibitory phosphorylation. Simultaneous expression of mutant proteins lacking these residues results in complete uncoupling from the cell cycle. Consequently, CENP-A assembly is fully recapitulated under high Cdk activities, indistinguishable from G1 assembly. We find that Cdk-mediated inhibition is exerted by sequestering active factors away from the centromere. Finally, we show that displacement of M18BP1 from the centromere is critical for the assembly mechanism of CENP-A.
authors.affiliation: Instituto Gulbenkian de Ciência, 2780-156 Oeiras, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Stankovic
authors.affiliation: Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Lucie Y
authors.initials: LY
authors.lastname: Guo
authors.affiliation: Instituto Gulbenkian de Ciência, 2780-156 Oeiras, Portugal.
authors.forename: João F
authors.initials: JF
authors.lastname: Mata
authors.affiliation: Instituto Gulbenkian de Ciência, 2780-156 Oeiras, Portugal.
authors.forename: Dani L
authors.initials: DL
authors.lastname: Bodor
authors.affiliation: Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Xing-Jun
authors.initials: XJ
authors.lastname: Cao
authors.affiliation: Department of Cell Biology, University of Virginia, Charlottesville, VA, 22901 USA.
authors.forename: Aaron O
authors.initials: AO
authors.lastname: Bailey
authors.affiliation: Department of Chemistry, University of Virginia, Charlottesville, VA, 22901 USA.
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Shabanowitz
authors.affiliation: Department of Chemistry, University of Virginia, Charlottesville, VA, 22901 USA.
authors.forename: Donald F
authors.initials: DF
authors.lastname: Hunt
authors.affiliation: Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Garcia
authors.affiliation: Epigenetics Program, Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA. Electronic address: blackbe@mail.med.upenn.edu.
authors.forename: Ben E
authors.initials: BE
authors.lastname: Black
authors.affiliation: Instituto Gulbenkian de Ciência, 2780-156 Oeiras, Portugal. Electronic address: ljansen@igc.gulbenkian.pt.
authors.forename: Lars E T
authors.initials: LET
authors.lastname: Jansen
mesh_terms: D001324:Autoantigens / Q000235:genetics / Q000378:metabolism*; D016203:CDC2 Protein Kinase; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D000076247:Centromere Protein A; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D042002:Chromatin Assembly and Disassembly; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D019925:Cyclin A / Q000235:genetics / Q000378:metabolism; D051357:Cyclin-Dependent Kinase 2 / Q000235:genetics / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D059585:G1 Phase Cell Cycle Checkpoints; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D015398:Signal Transduction; D014162:Transfection
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D019925:Cyclin A; D004268:DNA-Binding Proteins; C524371:HJURP protein, human; C517147:MIS18BP1 protein, human; D016203:CDC2 Protein Kinase; C528899:CDK1 protein, human; C495894:CDK2 protein, human; D051357:Cyclin-Dependent Kinase 2; D018844:Cyclin-Dependent Kinases
keywords: CENP-A; cell cycle; centromere; chromatin; cyclin dependent kinase; epigenetics; histone variant; kinetochore; mitosis