Acetylation of histone H4 lysine 5 and 12 is required for CENP-A deposition into centromeres.
Centromeres are specified epigenetically through the deposition of the centromere-specific histone H3 variant CENP-A. However, how additional epigenetic features are involved in centromere specification is unknown. Here, we find that histone H4 Lys5 and Lys12 acetylation (H4K5ac and H4K12ac) primarily occur within the pre-nucleosomal CENP-A-H4-HJURP (CENP-A chaperone) complex, before centromere deposition. We show that H4K5ac and H4K12ac are mediated by the RbAp46/48-Hat1 complex and that RbAp48-deficient DT40 cells fail to recruit HJURP to centromeres and do not incorporate new CENP-A at centromeres. However, C-terminally-truncated HJURP, that does not bind CENP-A, does localize to centromeres in RbAp48-deficient cells. Acetylation-dead H4 mutations cause mis-localization of the CENP-A-H4 complex to non-centromeric chromatin. Crucially, CENP-A with acetylation-mimetic H4 was assembled specifically into centromeres even in RbAp48-deficient DT40 cells. We conclude that H4K5ac and H4K12ac, mediated by RbAp46/48, facilitates efficient CENP-A deposition into centromeres.
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
Wei-Hao
WH
Shang
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
Tetsuya
T
Hori
Department of Biochemistry, Stanford University Medical School, 259 Campus Drive, Beckman B409, Stanford, California 94305, USA.
Frederick G
FG
Westhorpe
Department of Biochemistry, Geisel School of Medicine, Dartmouth College, HB7200, Hanover, New Hampshire 03755, USA.
Kristina M
KM
Godek
Comparative Genomics Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Atsushi
A
Toyoda
DNA Data Analysis Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Sadahiko
S
Misu
DNA Data Analysis Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Norikazu
N
Monma
DNA Data Analysis Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Kazuho
K
Ikeo
Department of Cell Biology, Yale University School of Medicine, SHM C-230, 333 Cedar St., New Haven, Connecticut 06520, USA.
Christopher W
CW
Carroll
Section of Biochemistry and Molecular Biology, Department of Medical Sciences, University of Miyazaki, 5200, Kihara, Kiyotake, Miyazaki 889-1692, Japan.
Yasunari
Y
Takami
Comparative Genomics Laboratory, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.|National Institute of Informatics, Hitotsubashi, Chiyoda-ku, Tokyo 101-8430, Japan.
Asao
A
Fujiyama
Cell Biology Unit, Institute of Innovative Research, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Hiroshi
H
Kimura
Department of Biochemistry, Stanford University Medical School, 259 Campus Drive, Beckman B409, Stanford, California 94305, USA.
Aaron F
AF
Straight
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
Tatsuo
T
Fukagawa
D000107:Acetylation; D000818:Animals; D045744:Cell Line, Tumor; D002503:Centromere / Q000235:genetics / Q000378:metabolism*; D000076247:Centromere Protein A / Q000235:genetics / Q000378:metabolism*; D002645:Chickens; D002843:Chromatin / Q000378:metabolism; D044127:Epigenesis, Genetic; D006657:Histones / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008239:Lysine / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D009707:Nucleosomes / Q000235:genetics / Q000378:metabolism*; D056505:Retinoblastoma-Binding Protein 4 / Q000378:metabolism; D056524:Retinoblastoma-Binding Protein 7 / Q000378:metabolism
C000619700:CENPA protein, human; D000076247:Centromere Protein A; D002843:Chromatin; D006657:Histones; D018832:Molecular Chaperones; D009707:Nucleosomes; C082637:RBBP4 protein, human; C490746:RBBP7 protein, human; D056505:Retinoblastoma-Binding Protein 4; D056524:Retinoblastoma-Binding Protein 7; D008239:Lysine