7BXT

The cryo-EM structure of CENP-A nucleosome in complex with CENP-C peptide and CENP-N N-terminal domain

Summary for 7BXT

• Entry DOI: 10.2210/pdb7bxt/pdb
• EMDB information: 30237
• Descriptor: Histone H3,Histone H3-like centromeric protein A, Histone H4, Histone H2A type 1-B/E, ... (8 entities in total)
• Functional Keywords: cenp-a nucleosome, cenp-c, cenp-n, complex, kinetochore, nuclear protein, cell cycle-dna complex, cell cycle/dna
• Biological source: Gallus gallus (Chicken); More
• Total number of polymer chains: 14
• Total formula weight: 355877.50

Authors

Ariyoshi, M.,Makino, F.,Fukagawa, T. (deposition date: 2020-04-20, release date: 2021-02-10, Last modification date: 2024-03-27)

Primary citation

Ariyoshi, M.,Makino, F.,Watanabe, R.,Nakagawa, R.,Kato, T.,Namba, K.,Arimura, Y.,Fujita, R.,Kurumizaka, H.,Okumura, E.I.,Hara, M.,Fukagawa, T.
Cryo-EM structure of the CENP-A nucleosome in complex with phosphorylated CENP-C.
Embo J., 40:e105671-e105671, 2021

PubMed 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.

PubMed: 33463726
DOI: 10.15252/embj.2020105671

Experimental method

ELECTRON MICROSCOPY (4.2 Å)