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

Research output: Contribution to journalArticle

117 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)794-805
Number of pages12
JournalMolecular Cell
Volume35
Issue number6
DOIs
StatePublished - Sep 24 2009

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Saccharomycetales
Nucleosomes
Dimerization
Histones
Chromatin
Micrococcal Nuclease
Chromosome Segregation
Proteins

Keywords

  • CELLCYCLE
  • DNA
  • PROTEINS

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Camahort, R., Shivaraju, M., Mattingly, M., Li, B., Nakanishi, S., Zhu, D., ... Gerton, J. L. (2009). Cse4 Is Part of an Octameric Nucleosome in Budding Yeast. Molecular Cell, 35(6), 794-805. https://doi.org/10.1016/j.molcel.2009.07.022

Cse4 Is Part of an Octameric Nucleosome in Budding Yeast. / Camahort, Raymond; Shivaraju, Manjunatha; Mattingly, Mark; Li, Bing; Nakanishi, Shima; Zhu, Dongxiao; Shilatifard, Ali; Workman, Jerry L.; Gerton, Jennifer L.

In: Molecular Cell, Vol. 35, No. 6, 24.09.2009, p. 794-805.

Research output: Contribution to journalArticle

Camahort, R, Shivaraju, M, Mattingly, M, Li, B, Nakanishi, S, Zhu, D, Shilatifard, A, Workman, JL & Gerton, JL 2009, 'Cse4 Is Part of an Octameric Nucleosome in Budding Yeast', Molecular Cell, vol. 35, no. 6, pp. 794-805. https://doi.org/10.1016/j.molcel.2009.07.022
Camahort R, Shivaraju M, Mattingly M, Li B, Nakanishi S, Zhu D et al. Cse4 Is Part of an Octameric Nucleosome in Budding Yeast. Molecular Cell. 2009 Sep 24;35(6):794-805. https://doi.org/10.1016/j.molcel.2009.07.022
Camahort, Raymond ; Shivaraju, Manjunatha ; Mattingly, Mark ; Li, Bing ; Nakanishi, Shima ; Zhu, Dongxiao ; Shilatifard, Ali ; Workman, Jerry L. ; Gerton, Jennifer L. / Cse4 Is Part of an Octameric Nucleosome in Budding Yeast. In: Molecular Cell. 2009 ; Vol. 35, No. 6. pp. 794-805.
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