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Molecular cell
Sep 24, 2009;35 (6): 794-805.

Cse4 is part of an octameric nucleosome in budding yeast.

Raymond Camahort, Manjunatha Shivaraju, Mark Mattingly, Bing Li, Shima Nakanishi, Dongxiao Zhu, Ali Shilatifard, Jerry L Workman, Jennifer L Gerton
Author Information
  1. Raymond Camahort: Stowers Institute for Medical Research, Kansas City, MO 64110, USA.
PMID: 19782029 DOI: 10.1016/j.molcel.2009.07.022

Abstract

The budding yeast CenH3 histone variant Cse4 localizes to centromeric nucleosomes and is required for kinetochore assembly and chromosome segregation. The exact composition of centromeric Cse4-containing nucleosomes is a subject of debate. Using unbiased biochemical, cell-biological, and genetic approaches, we have tested the composition of Cse4-containing nucleosomes. Using micrococcal nuclease-treated chromatin, we find that Cse4 is associated with the histones H2A, H2B, and H4, but not H3 or the nonhistone protein Scm3. Overexpression of Cse4 rescues the lethality of a scm3 deletion, indicating that Scm3 is not essential for the formation of functional centromeric chromatin. We also find that octameric Cse4 nucleosomes can be reconstituted in vitro. Furthermore, Cse4-Cse4 dimerization occurs in vivo at the centromeric nucleosome, and this requires the predicted Cse4-Cse4 dimerization interface. Taken together, our experimental evidence supports the model that the Cse4 nucleosome is an octamer, containing two copies each of Cse4, H2A, H2B, and H4.

Associated Data

GEO | GSE17481

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Grants

  1. R01 CA089455/NCI NIH HHS
  2. R01 GM080477/NIGMS NIH HHS
  3. R01 GM080477-01A2/NIGMS NIH HHS
  4. R01GM080477/NIGMS NIH HHS

MeSH Term

Centromere
Chromatin Immunoprecipitation
Chromosomal Proteins, Non-Histone
DNA-Binding Proteins
Gene Expression Regulation, Fungal
Histones
Models, Molecular
Multiprotein Complexes
Mutation
Nucleic Acid Conformation
Nucleosomes
Protein Conformation
Protein Multimerization
Protein Subunits
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins

Chemicals

CSE4 protein, S cerevisiae
Chromosomal Proteins, Non-Histone
DNA-Binding Proteins
Histones
Multiprotein Complexes
Nucleosomes
Protein Subunits
Saccharomyces cerevisiae Proteins
Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't
Article has an altmetric score of 1

Word Cloud

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