title: Epigenetic engineering shows that a human centromere resists silencing mediated by H3K27me3/K9me3.
abstract: Centromeres are characterized by the centromere-specific H3 variant CENP-A, which is embedded in chromatin with a pattern characteristic of active transcription that is required for centromere identity. It is unclear how centromeres remain transcriptionally active despite being flanked by repressive pericentric heterochromatin. To further understand centrochromatin's response to repressive signals, we nucleated a Polycomb-like chromatin state within the centromere of a human artificial chromosome (HAC) by tethering the methyltransferase EZH2. This led to deposition of the H3K27me3 mark and PRC1 repressor binding. Surprisingly, this state did not abolish HAC centromere function or transcription, and this apparent resistance was not observed on a noncentromeric locus, where transcription was silenced. Directly tethering the reader/repressor PRC1 bypassed this resistance, inactivating the centromere. We observed analogous responses when tethering the heterochromatin Editor Suv39h1-methyltransferase domain (centromere resistance) or reader HP1α (centromere inactivation), respectively. Our results reveal that the HAC centromere can resist repressive pathways driven by H3K9me3/H3K27me3 and may help to explain how centromeres are able to resist inactivation by flanking heterochromatin.
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland, United Kingdom.
authors.forename: Nuno M C
authors.initials: NM
authors.lastname: Martins
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland, United Kingdom.
authors.forename: Jan H
authors.initials: JH
authors.lastname: Bergmann
authors.affiliation: Laboratory of Cell Engineering, Department of Frontier Research, Kazusa DNA Research Institute, Kisarazu 292-0818, Japan Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya 464-8602, Japan.
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Shono
authors.affiliation: Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kimura
authors.affiliation: Laboratory of Molecular Pharmacology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Larionov
authors.affiliation: Laboratory of Cell Engineering, Department of Frontier Research, Kazusa DNA Research Institute, Kisarazu 292-0818, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Masumoto
authors.affiliation: Wellcome Trust Centre for Cell Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland, United Kingdom bill.earnshaw@ed.ac.uk.
authors.forename: William C
authors.initials: WC
authors.lastname: Earnshaw
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism; D002478:Cells, Cultured; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D002843:Chromatin / Q000378:metabolism; D000090266:Chromobox Protein Homolog 5; D022222:Chromosomes, Artificial, Human; D000071221:Enhancer of Zeste Homolog 2 Protein; D044127:Epigenesis, Genetic; D057890:Epigenomics / Q000379:methods; D006367:HeLa Cells; D006570:Heterochromatin / Q000378:metabolism; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008780:Methyltransferases / Q000235:genetics; D063151:Polycomb Repressive Complex 2
chemical_list: C000715594:CBX5 protein, human; D018797:Cell Cycle Proteins; D002843:Chromatin; D006570:Heterochromatin; D006657:Histones; C116662:PRC1 protein, human; D000090266:Chromobox Protein Homolog 5; D008780:Methyltransferases; C498409:EZH2 protein, human; D000071221:Enhancer of Zeste Homolog 2 Protein; D063151:Polycomb Repressive Complex 2