TY - JOUR
T1 - The centromeric nucleosome-like CENP-T-W-S-X complex induces positive supercoils into DNA
AU - Takeuchi, Kozo
AU - Nishino, Tatsuya
AU - Mayanagi, Kouta
AU - Horikoshi, Naoki
AU - Osakabe, Akihisa
AU - Tachiwana, Hiroaki
AU - Hori, Tetsuya
AU - Kurumizaka, Hitoshi
AU - Fukagawa, Tatsuo
N1 - Funding Information:
A grant-in-Aid for Scientific Research (S) from the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (to T.F.); Cabinet Office, Government of Japan [Funding Program for Next Generation World-Leading Researchers to T.F.]; The International Human Frontier Science Program Organization (to T.N.); Grants-in-Aid for Scientific Research from MEXT (to T.N., T.H. and K.M.). PRESTO of JST (to T.H. and K.M.). Funding for open access charge: A grant-in-Aid for Scientific Research (S) from MEXT, Japan (to T.F.).
PY - 2014/2
Y1 - 2014/2
N2 - 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.
AB - 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.
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U2 - 10.1093/nar/gkt1124
DO - 10.1093/nar/gkt1124
M3 - Article
C2 - 24234442
AN - SCOPUS:84896730017
SN - 0305-1048
VL - 42
SP - 1644
EP - 1655
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 3
ER -