CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore.
Kinetochore specification and assembly requires the targeted deposition of specialized nucleosomes containing the histone H3 variant CENP-A at centromeres. However, CENP-A is not sufficient to drive full-kinetochore assembly, and it is not clear how centromeric chromatin is established. Here, we identify CENP-W as a component of the DNA-proximal constitutive centromere-associated network (CCAN) of proteins. We demonstrate that CENP-W forms a DNA-binding complex together with the CCAN component CENP-T. This complex directly associates with nucleosomal DNA and with canonical histone H3, but not with CENP-A, in centromeric regions. CENP-T/CENP-W functions upstream of other CCAN components with the exception of CENP-C, an additional putative DNA-binding protein. Our analysis indicates that CENP-T/CENP-W and CENP-C provide distinct pathways to connect the centromere with outer kinetochore assembly. In total, our results suggest that the CENP-T/CENP-W complex is directly involved in establishment of centromere chromatin structure coordinately with CENP-A.
Hori
Tetsuya
T
Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan.
Amano
Miho
M
Suzuki
Aussie
A
Backer
Chelsea B
CB
Welburn
Julie P
JP
Dong
Yimin
Y
McEwen
Bruce F
BF
Shang
Wei-Hao
WH
Suzuki
Emiko
E
Okawa
Katsuya
K
Cheeseman
Iain M
IM
Fukagawa
Tatsuo
T
D000595:Amino Acid Sequence; D000818:Animals; D001324:Autoantigens / Q000235:genetics / Q000378:metabolism; D002503:Centromere; D000076247:Centromere Protein A; D002645:Chickens; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D009154:Mutation; D009707:Nucleosomes / Q000378:metabolism
D001324:Autoantigens; C000619700:CENPA protein, human; C534001:CENPT protein, human; C534000:CENPW protein, human; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D009707:Nucleosomes; D004247:DNA