Cryo-EM Structure of the CENP-A Nucleosome in Complex with Phosphorylated CENP-C

Overview

Journal EMBO J

Specialties Biotechnology
Molecular Biology

Date 2021 Jan 19

PMID 33463726

Citations 19

Authors

Mariko Ariyoshi

Fumiaki Makino

Reito Watanabe

Reiko Nakagawa

Takayuki Kato

Keiichi Namba

Yasuhiro Arimura

Risa Fujita

Hitoshi Kurumizaka

Ei-Ichi Okumura

Masatoshi Hara

Tatsuo Fukagawa

Affiliations

Soon will be listed here.

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.

Citing Articles

Non-nucleosomal (CENP-A/H4) - DNA complexes as a possible platform for centromere organization.

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Centromeric localization of αKNL2 and CENP-C proteins in plants depends on their centromere-targeting domain and DNA-binding regions.

Yalagapati S, Ahmadli U, Sinha A, Kalidass M, Dabravolski S, Zuo S Nucleic Acids Res. 2024; 53(4).

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CENcyclopedia: Dynamic Landscape of Kinetochore Architecture Throughout the Cell Cycle.

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Simultaneous profiling of chromatin accessibility and DNA methylation in complete plant genomes using long-read sequencing.

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CENP-A and CENP-B collaborate to create an open centromeric chromatin state.

Nagpal H, Ali-Ahmad A, Hirano Y, Cai W, Halic M, Fukagawa T Nat Commun. 2023; 14(1):8227.

PMID: 38086807 PMC: 10716449. DOI: 10.1038/s41467-023-43739-5.

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