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Updated: May 12, 2026
05:35
Immunofluorescence Analysis of Endogenous and Exogenous Centromere-kinetochore Proteins
Published on: March 3, 2016
Histone H3 localizes to the centromeric DNA in budding yeast
Berit Lochmann1, Dmitri Ivanov
1Friedrich Miescher Laboratory of the Max Planck Society, Tübingen, Germany.
Plos Genetics
|June 14, 2012
Summary
Budding yeast centromeres contain both histone H3 and the Cse4 variant (CENP-A) within a single nucleosome. This study used ChIP to reveal this H3-Cse4 heterotypic octamer composition at centromeric DNA.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Kinetochores mediate sister chromatid segregation during cell division.
- Centromeric DNA organizes kinetochores via specialized nucleosomes.
- Budding yeast centromeres are uniquely defined by a 125 bp sequence and a single nucleosome.
Purpose of the Study:
- To investigate the precise composition of the centromeric nucleosome in budding yeast.
- To determine the localization of histone variants and chaperones at centromeric DNA.
Main Methods:
- Novel Chromatin Immunoprecipitation (ChIP) approach.
- Sequential ChIP to assess co-occupancy of proteins.
- Analysis of protein interactions with centromeric DNA throughout the cell cycle.
Main Results:
- Histone H3, Cse4 (CENP-A), H4, H2A, and H2B interact with centromeric DNA.
- Histone H3 is localized to centromeric DNA across all cell cycle stages.
- Co-occupancy of H3 and Cse4 at centromeric DNA was demonstrated, suggesting a H3-Cse4 heterotypic octamer.
Conclusions:
- The centromeric nucleosome in budding yeast is likely a heterotypic octamer composed of H3 and Cse4 (CENP-A).
- Histone H3 plays a stable and integral role at the centromeric DNA, alongside the Cse4 variant.
- Future models of centromere composition must incorporate the stable association of histone H3.
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