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Histone-histone interactions and centromere function.

L Glowczewski1, P Yang, T Kalashnikova

  • 1Department of Microbiology and Cancer Center, University of Virginia, Charlottesville, Virginia 22908, USA.

Molecular and Cellular Biology
|July 13, 2000
PubMed
Summary
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The study confirms that Cse4p and histone H4 interact to form centromere nucleosomes in yeast. This interaction is crucial for proper chromosome segregation and centromere function.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Cse4p is a specialized histone H3 variant essential for centromere structure and function in Saccharomyces cerevisiae.
  • Histone H4 mutations (hhf1-20) cause defects in centromere chromatin and chromosome transmission.
  • A model proposes Cse4p and histone H4 interact via their histone fold domains to form centromeric nucleosomes.

Purpose of the Study:

  • To genetically test the proposed interaction between Cse4p and histone H4.
  • To identify specific regions of Cse4p involved in this interaction.
  • To investigate the role of histone H3 in centromere assembly.

Main Methods:

  • Targeted random mutagenesis of the Cse4p histone fold domain.
  • Isolation and characterization of temperature-sensitive cse4 alleles.

Related Experiment Videos

  • Genetic analysis including allele-specific suppression and dosage lethality experiments.

    • Main Results:

    • Three temperature-sensitive cse4 alleles were identified, two with mutations at the Cse4p-H4 interface, one showing long-range cooperative effects.
    • A third allele had mutations in the helix 2-helix 3 interface, important for H3 dimerization.
    • Reciprocal allele-specific suppression between Cse4p and histone H4 overexpression confirmed their interaction.
    • Histone H3 overexpression was lethal in cse4 mutants, indicating competition for histone H4.

    Conclusions:

    • The results provide strong genetic evidence for a direct Cse4p-histone H4 interaction in centromere assembly.
    • Centromere chromatin assembly is a highly regulated process, evidenced by the pathway's resistance to histone H3 interference.
    • Understanding Cse4p-H4 interactions is key to comprehending centromere structure and epigenetic inheritance.