title: CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore.
abstract: 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.
authors.affiliation: Department of Molecular Genetics, National Institute of Genetics and The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Hori
authors.forename: Miho
authors.initials: M
authors.lastname: Amano
authors.forename: Aussie
authors.initials: A
authors.lastname: Suzuki
authors.forename: Chelsea B
authors.initials: CB
authors.lastname: Backer
authors.forename: Julie P
authors.initials: JP
authors.lastname: Welburn
authors.forename: Yimin
authors.initials: Y
authors.lastname: Dong
authors.forename: Bruce F
authors.initials: BF
authors.lastname: McEwen
authors.forename: Wei-Hao
authors.initials: WH
authors.lastname: Shang
authors.forename: Emiko
authors.initials: E
authors.lastname: Suzuki
authors.forename: Katsuya
authors.initials: K
authors.lastname: Okawa
authors.forename: Iain M
authors.initials: IM
authors.lastname: Cheeseman
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Fukagawa
mesh_terms: 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
chemical_list: 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