title: Histone H4 Lys 20 monomethylation of the CENP-A nucleosome is essential for kinetochore assembly.
abstract: In vertebrate cells, centromeres are specified epigenetically through the deposition of the centromere-specific histone CENP-A. Following CENP-A deposition, additional proteins are assembled on centromeric chromatin. However, it remains unknown whether additional epigenetic features of centromeric chromatin are required for kinetochore assembly. Here, we used ChIP-seq analysis to examine centromere-specific histone modifications at chicken centromeres, which lack highly repetitive sequences. We found that H4K20 monomethylation (H4K20me1) is enriched at centromeres. Immunofluorescence and biochemical analyses revealed that H4K20me1 is present at all centromeres in chicken and human cells. Based on immunoprecipitation data, H4K20me1 occurs primarily on the histone H4 that is assembled as part of the CENP-A nucleosome following deposition of CENP-A into centromeres. Targeting the H4K20me1-specific demethylase PHF8 to centromeres reduces the level of H4K20me1 at centromeres and results in kinetochore assembly defects. We conclude that H4K20me1 modification of CENP-A nucleosomes contributes to functional kinetochore assembly.
authors: Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan.
authors: Tetsuya
authors: T
authors: Hori
authors: Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan.
authors: Wei-Hao
authors: WH
authors: Shang
authors: Comparative Genomics Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors: Atsushi
authors: A
authors: Toyoda
authors: Cell Innovation Project, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors: Sadahiko
authors: S
authors: Misu
authors: Cell Innovation Project, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors: Norikazu
authors: N
authors: Monma
authors: Cell Innovation Project, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
authors: Kazuho
authors: K
authors: Ikeo
authors: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
authors: Oscar
authors: O
authors: Molina
authors: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
authors: Giulia
authors: G
authors: Vargiu
authors: Comparative Genomics Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan; National Institute of Informatics, Hitotsubashi, Chiyoda-ku, Tokyo 101-8430, Japan.
authors: Asao
authors: A
authors: Fujiyama
authors: Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamada-oka, Suita, Osaka 565-0871, Japan.
authors: Hiroshi
authors: H
authors: Kimura
authors: Wellcome Trust Centre for Cell Biology, University of Edinburgh, King's Buildings, Mayfield Road, Edinburgh EH9 3JR, UK.
authors: William C
authors: WC
authors: Earnshaw
authors: Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan. Electronic address: tfukagaw@lab.nig.ac.jp.
authors: Tatsuo
authors: T
authors: Fukagawa
mesh_terms: D000818:Animals; D001324:Autoantigens / Q000378:metabolism*; D002503:Centromere / Q000378:metabolism; D000076247:Centromere Protein A; D002645:Chickens; D042002:Chromatin Assembly and Disassembly; D047369:Chromatin Immunoprecipitation; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D019175:DNA Methylation; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D008239:Lysine / Q000378:metabolism*; D009707:Nucleosomes / Q000378:metabolism*
chemical_list: D001324:Autoantigens; C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D009707:Nucleosomes; D008239:Lysine