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Nucleic Acids Res. 2014 Feb;42(3):1644-55. doi: 10.1093/nar/gkt1124. Epub 2013 Nov 14.

The centromeric nucleosome-like CENP-T-W-S-X complex induces positive supercoils into DNA.

Author information

1
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.

Abstract

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.

PMID:
24234442
PMCID:
PMC3919578
DOI:
10.1093/nar/gkt1124
[Indexed for MEDLINE]
Free PMC Article
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