title: Epigenetic engineering shows H3K4me2 is required for HJURP targeting and CENP-A assembly on a synthetic human kinetochore.
abstract: Kinetochores assemble on distinct 'centrochromatin' containing the histone H3 variant CENP-A and interspersed nucleosomes dimethylated on H3K4 (H3K4me2). Little is known about how the chromatin environment at active centromeres governs centromeric structure and function. Here, we report that centrochromatin resembles K4-K36 domains found in the body of some actively transcribed housekeeping genes. By tethering the lysine-specific demethylase 1 (LSD1), we specifically depleted H3K4me2, a modification thought to have a role in transcriptional memory, from the kinetochore of a synthetic human artificial chromosome (HAC). H3K4me2 depletion caused kinetochores to suffer a rapid loss of transcription of the underlying α-satellite DNA and to no longer efficiently recruit HJURP, the CENP-A chaperone. Kinetochores depleted of H3K4me2 remained functional in the short term, but were defective in incorporation of CENP-A, and were gradually inactivated. Our data provide a functional link between the centromeric chromatin, α-satellite transcription, maintenance of CENP-A levels and kinetochore stability.
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh, Scotland, UK.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Bergmann
authors.forename: Mariluz Gómez
authors.initials: MG
authors.lastname: Rodríguez
authors.forename: Nuno M C
authors.initials: NM
authors.lastname: Martins
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.forename: David A
authors.initials: DA
authors.lastname: Kelly
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Masumoto
authors.forename: Vladimir
authors.initials: V
authors.lastname: Larionov
authors.forename: Lars E T
authors.initials: LE
authors.lastname: Jansen
authors.forename: William C
authors.initials: WC
authors.lastname: Earnshaw
mesh_terms: D001324:Autoantigens / Q000378:metabolism*; D002503:Centromere / Q000378:metabolism / Q000502:physiology*; D000076247:Centromere Protein A; D002843:Chromatin / Q000235:genetics / Q000378:metabolism*; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D022222:Chromosomes, Artificial, Human / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D004268:DNA-Binding Proteins / Q000378:metabolism*; D044127:Epigenesis, Genetic / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods; D006657:Histones / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D017931:DNA Primers; D004268:DNA-Binding Proteins; C524371:HJURP protein, human; D006657:Histones; D009707:Nucleosomes