6UPH

Structure of a Yeast Centromeric Nucleosome at 2.7 Angstrom resolution

Summary for 6UPH

• Entry DOI: 10.2210/pdb6uph/pdb
• EMDB information: 20839
• Descriptor: Histone H3-like centromeric protein CSE4, Histone H4, Histone H2A, ... (6 entities in total)
• Functional Keywords: histones, nucleosome, centromere, kinetochore, yeast, cell cycle
• Biological source: Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast); More
• Total number of polymer chains: 10
• Total formula weight: 235101.28

Authors

Migl, D.,Kschonsak, M.,Arthur, C.P.,Khin, Y.,Harrison, S.C.,Ciferri, C.,Dimitrova, Y.N. (deposition date: 2019-10-17, release date: 2019-11-06, Last modification date: 2025-05-14)

Primary citation

Migl, D.,Kschonsak, M.,Arthur, C.P.,Khin, Y.,Harrison, S.C.,Ciferri, C.,Dimitrova, Y.N.
Cryoelectron Microscopy Structure of a Yeast Centromeric Nucleosome at 2.7 angstrom Resolution.
Structure, 28:363-370.e3, 2020

PubMed Abstract: Kinetochores mediate chromosome segregation during cell division. They assemble on centromeric nucleosomes and capture spindle microtubules. In budding yeast, a kinetochore links a single nucleosome, containing the histone variant Cse4 instead of H3, with a single microtubule. Conservation of most kinetochore components from yeast to metazoans suggests that the yeast kinetochore represents a module of the more complex metazoan arrangements. We describe here a streamlined protocol for reconstituting a yeast centromeric nucleosome and a systematic exploration of cryo-grid preparation. These developments allowed us to obtain a high-resolution cryoelectron microscopy reconstruction. As suggested by previous work, fewer base pairs are in tight association with the histone octamer than there are in canonical nucleosomes. Weak binding of the end DNA sequences may contribute to specific recognition by other inner kinetochore components. The centromeric nucleosome structure and the strategies we describe will facilitate studies of many other aspects of kinetochore assembly and chromatin biochemistry.

PubMed: 32004465
DOI: 10.1016/j.str.2019.12.002

Experimental method

ELECTRON MICROSCOPY (2.7 Å)