The centromeric nucleosome-like CENP-T-W-S-X complex induces positive supercoils into DNA.
The centromere is a specific genomic region upon which the kinetochore is formed to attach to spindle microtubules for faithful chromosome segregation. To distinguish this chromosomal region from other genomic loci, the centromere contains a specific chromatin structure including specialized nucleosomes containing the histone H3 variant CENP-A. In addition to CENP-A nucleosomes, we have found that centromeres contain a nucleosome-like structure comprised of the histone-fold CENP-T-W-S-X complex. However, it is unclear how the CENP-T-W-S-X complex associates with centromere chromatin. Here, we demonstrate that the CENP-T-W-S-X complex binds preferentially to ∼ 100 bp of linker DNA rather than nucleosome-bound DNA. In addition, we find that the CENP-T-W-S-X complex primarily binds to DNA as a (CENP-T-W-S-X)2 structure. Interestingly, in contrast to canonical nucleosomes that negatively supercoil DNA, the CENP-T-W-S-X complex induces positive DNA supercoils. We found that the DNA-binding regions in CENP-T or CENP-W, but not CENP-S or CENP-X, are required for this positive supercoiling activity and the kinetochore targeting of the CENP-T-W-S-X complex. In summary, our work reveals the structural features and properties of the CENP-T-W-S-X complex for its localization to centromeres.
Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan, The Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Fukuoka 812-8581, Japan, JST, PRESTO, Fukuoka 812-8582, Japan and Laboratory of Structural Biology, Graduate School of Advanced Science and Engineering, Waseda University, Shinjuku-ku, Tokyo 162-8480, Japan.
Kozo
K
Takeuchi
Tatsuya
T
Nishino
Kouta
K
Mayanagi
Naoki
N
Horikoshi
Akihisa
A
Osakabe
Hiroaki
H
Tachiwana
Tetsuya
T
Hori
Hitoshi
H
Kurumizaka
Tatsuo
T
Fukagawa
D000818:Animals; D002460:Cell Line; D002503:Centromere / Q000378:metabolism*; D002645:Chickens; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D004278:DNA, Superhelical / Q000378:metabolism*; D018386:Kinetochores / Q000378:metabolism; D009707:Nucleosomes / Q000378:metabolism
D002868:Chromosomal Proteins, Non-Histone; D004278:DNA, Superhelical; D009707:Nucleosomes; D004247:DNA