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Cryo-EM structure of the CENP-A nucleosome in complex with phosphorylated CENP-C.
Ariyoshi, Mariko; Makino, Fumiaki; Watanabe, Reito; Nakagawa, Reiko; Kato, Takayuki; Namba, Keiichi; Arimura, Yasuhiro; Fujita, Risa; Kurumizaka, Hitoshi; Okumura, Ei-Ichi; Hara, Masatoshi; Fukagawa, Tatsuo.
Afiliação
  • Ariyoshi M; Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
  • Makino F; Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
  • Watanabe R; JEOL Ltd., Akishima, Tokyo, Japan.
  • Nakagawa R; Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
  • Kato T; Laboratory for Phyloinformatics, RIKEN Center for Biosystems Dynamics Research (BDR), Kobe, Hyogo, Japan.
  • Namba K; Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
  • Arimura Y; Institute of Protein Research, Osaka University, Suita, Osaka, Japan.
  • Fujita R; Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, Japan.
  • Kurumizaka H; RIKEN Center for Biosystems Dynamics Research (BDR) and SPring-8 Center, and JEOL YOKOGUSHI Research Alliance Laboratories, Osaka University, Suita, Osaka, Japan.
  • Okumura EI; Laboratory of Chromatin Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan.
  • Hara M; Laboratory of Chromatin Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan.
  • Fukagawa T; Laboratory of Chromatin Structure and Function, Institute for Quantitative Biosciences, The University of Tokyo, Tokyo, Japan.
EMBO J ; 40(5): e105671, 2021 03 01.
Article em En | MEDLINE | ID: mdl-33463726
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Cromossômicas não Histona / Nucleossomos / Centrômero / Cinetocoros / Microscopia Crioeletrônica / Proteína Centromérica A Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Revista: EMBO J Ano de publicação: 2021 Tipo de documento: Article
Texto completo
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Proteínas Cromossômicas não Histona / Nucleossomos / Centrômero / Cinetocoros / Microscopia Crioeletrônica / Proteína Centromérica A Tipo de estudo: Prognostic_studies Limite: Animals Idioma: En Revista: EMBO J Ano de publicação: 2021 Tipo de documento: Article