Dynamic changes in CCAN organization through CENP-C during cell-cycle progression.
The kinetochore is a crucial structure for faithful chromosome segregation during mitosis and is formed in the centromeric region of each chromosome. The 16-subunit protein complex known as the constitutive centromere-associated network (CCAN) forms the foundation for kinetochore assembly on the centromeric chromatin. Although the CCAN can be divided into several subcomplexes, it remains unclear how CCAN proteins are organized to form the functional kinetochore. In particular, this organization may vary as the cell cycle progresses. To address this, we analyzed the relationship of centromeric protein (CENP)-C with the CENP-H complex during progression of the cell cycle. We find that the middle portion of chicken CENP-C (CENP-C(166-324)) is sufficient for centromere localization during interphase, potentially through association with the CENP-L-N complex. The C-terminus of CENP-C (CENP-C(601-864)) is essential for centromere localization during mitosis, through binding to CENP-A nucleosomes, independent of the CENP-H complex. On the basis of these results, we propose that CCAN organization changes dynamically during progression of the cell cycle.
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan Department of Molecular Genetics, National Institute of Genetics and Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan.
Harsh
H
Nagpal
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
Tetsuya
T
Hori
Bioorganic Research Institute, Suntory Foundation for Life Sciences, Osaka 618-8503, Japan.
Ayako
A
Furukawa
Bioorganic Research Institute, Suntory Foundation for Life Sciences, Osaka 618-8503, Japan.
Kenji
K
Sugase
Graduate School of Advanced Science and Engineering, Waseda University, Shinjuku, Tokyo 162-8480, Japan.
Akihisa
A
Osakabe
Graduate School of Advanced Science and Engineering, Waseda University, Shinjuku, Tokyo 162-8480, Japan.
Hitoshi
H
Kurumizaka
Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan Department of Molecular Genetics, National Institute of Genetics and Graduate University for Advanced Studies (SOKENDAI), Mishima, Shizuoka 411-8540, Japan tfukagawa@fbs.osaka-u.ac.jp tfukagaw@lab.nig.ac.jp.
Tatsuo
T
Fukagawa
D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D002503:Centromere / Q000378:metabolism; D002645:Chickens; D002843:Chromatin / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D007399:Interphase / Q000502:physiology; D018386:Kinetochores / Q000378:metabolism; D008938:Mitosis / Q000502:physiology; D009707:Nucleosomes / Q000378:metabolism; D017434:Protein Structure, Tertiary
D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D009707:Nucleosomes; C060905:centromere protein C