title: Breaking the HAC Barrier: histone H3K9 acetyl/methyl balance regulates CENP-A assembly.
abstract: The kinetochore is responsible for accurate chromosome segregation. However, the mechanism by which kinetochores assemble and are maintained remains unclear. Here we report that de novo CENP-A assembly and kinetochore formation on human centromeric alphoid DNA arrays is regulated by a histone H3K9 acetyl/methyl balance. Tethering of histone acetyltransferases (HATs) to alphoid DNA arrays breaks a cell type-specific barrier for de novo stable CENP-A assembly and induces assembly of other kinetochore proteins at the ectopic alphoid site. Similar results are obtained following tethering of CENP-A deposition factors hMis18α or HJURP. HAT tethering bypasses the need for hMis18α, but HJURP is still required for de novo kinetochore assembly. In contrast, H3K9 methylation following tethering of H3K9 tri-methylase (Suv39h1) to the array prevents de novo CENP-A assembly and kinetochore formation. CENP-A arrays assembled de novo by this mechanism can form human artificial chromosomes (HACs) that are propagated indefinitely in human cells.
authors.affiliation: Department of Human Genome Research, Laboratory of Cell Engineering, Kazusa DNA Research Institute, Kisarazu, Chiba, Japan.
authors.forename: Jun-ichirou
authors.initials: J
authors.lastname: Ohzeki
authors.forename: Jan H
authors.initials: JH
authors.lastname: Bergmann
authors.forename: Natalay
authors.initials: N
authors.lastname: Kouprina
authors.forename: Vladimir N
authors.initials: VN
authors.lastname: Noskov
authors.forename: Megumi
authors.initials: M
authors.lastname: Nakano
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.forename: William C
authors.initials: WC
authors.lastname: Earnshaw
authors.forename: Vladimir
authors.initials: V
authors.lastname: Larionov
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Masumoto
mesh_terms: D000107:Acetylation; D001324:Autoantigens / Q000378:metabolism*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D008745:Methylation; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D004247:DNA