SEARCH

PAGETOP

Cryo-EM structure of the CENP-A nucleosome in complex with phosphorylated CENP-C

Journal Embo J. e105671 (2021)
Title Cryo-EM structure of the CENP-A nucleosome in complex with phosphorylated CENP-C
Laboratory Laboratory of Chromosome Biology〈Prof. FUKAGAWA Tatsuo〉
Abstract

The CENP-A nucleosome is a key structure for kinetochore assembly. Once the CENP-A nucleosome is established in the centromere, additional proteins recognize the CENP-A nucleosome to form a kinetochore. CENP-C and CENP-N are CENP-A binding proteins. We previously demonstrated that vertebrate CENP-C binding to the CENP-A nucleosome is regulated by CDK1-mediated CENP-C phosphorylation. However, it is still unknown how the phosphorylation of CENP-C regulates its binding to CENP-A. It is also not completely understood how and whether CENP-C and CENP-N act together on the CENP-A nucleosome. Here, using cryo-electron microscopy (cryo-EM) in combination with biochemical approaches, we reveal a stable CENP-A nucleosome-binding mode of CENP-C through unique regions. The chicken CENP-C structure bound to the CENP-A nucleosome is stabilized by an intramolecular link through the phosphorylated CENP-C residue. The stable CENP-A-CENP-C complex excludes CENP-N from the CENP-A nucleosome. These findings provide mechanistic insights into the dynamic kinetochore assembly regulated by CDK1-mediated CENP-C phosphorylation.

Authors

Mariko Ariyoshi (1), Fumiaki Makino (1, 2), Reito Watanabe (1), Reiko Nakagawa (3), Takayuki Kato (1, 4), Keiichi Namba (1, 5), Yasuhiro Arimura (6), Risa Fujita (6), Hitoshi Kurumizaka (6), Ei‐ichi Okumura (7), Masatoshi Hara (1), Tatsuo Fukagawa (1)


  1. Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan
  2. JEOL Ltd., Akishima, Tokyo, Japan
  3. Laboratory for Phyloinformatics, RIKEN Center for Biosystems Dynamics Research (BDR), Kobe, Hyogo, Japan
  4. Institute of Protein Research, Osaka University, Suita, Osaka, Japan
  5. RIKEN Center for Biosystems Dynamics Research (BDR) and SPring‐8 Center, and JEOL YOKOGUSHI Research Alliance Laboratories, Osaka University, Suita, Osaka, Japan
  6. Laboratory of Chromatin Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan
  7. Laboratory of Cell and Developmental Biology, Graduate School of Bioscience, Tokyo Institute of Technology, Yokohama, Japan
PubMed 33463726

PAGETOP