Ctf3p, the Mis6 budding yeast homolog, interacts with Mcm22p and Mcm16p at the yeast outer kinetochore
Copyright policy )Ctf3p, the Mis6 budding yeast homolog, interacts with Mcm22p and Mcm16p at the yeast outer kinetochore
The budding yeast kinetochore is composed of an inner and outer protein complex, which binds to centromere (CEN) DNA and attaches to microtubules. We performed a genetic synthetic dosage lethality screen to identify novel kinetochore proteins in a collection of chromosome transmission fidelity mutants. Our screen identified several new kinetochore-related proteins including YLR381Wp/Ctf3p, which is a member of a conserved family of centromere-binding proteins. Ctf3p interacts with Mcm22p, Mcm16p, and the outer kinetochore protein Ctf19p. We used chromatin immunoprecipitation to demonstrate that Ctf3p, Mcm22p, and Mcm16p bind to CEN DNA in a Ctf19p-dependent manner. In addition, Ctf3p, Mcm22p, and Mcm16p have a localization pattern similar to other kinetochore proteins. The fission yeast Ctf3p homolog, Mis6, is required for loading of a CENP-A centromere specific histone, Cnp1, onto centromere DNA. We find however that Ctf3p is not required for loading of the budding yeast CENP-A homolog, Cse4p, ontoCEN DNA. In contrast, Ctf3p and Ctf19p fail to bind properly to the centromere in a cse4-1 mutant strain. We conclude that the requirements for CENP-A loading onto centromere DNA differ in fission versus budding yeast.
- University of British Columbia Canada
- University of Washington United States
- University of Toronto Canada
- University of Mary United States
- University of California, San Francisco United States
Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, Molecular Sequence Data, Cell Cycle Proteins, Saccharomyces cerevisiae, DNA-Binding Proteins, Fungal Proteins, Chromosome Segregation, Mutation, Amino Acid Sequence, Schizosaccharomyces pombe Proteins, Kinetochores
Saccharomyces cerevisiae Proteins, Sequence Homology, Amino Acid, Molecular Sequence Data, Cell Cycle Proteins, Saccharomyces cerevisiae, DNA-Binding Proteins, Fungal Proteins, Chromosome Segregation, Mutation, Amino Acid Sequence, Schizosaccharomyces pombe Proteins, Kinetochores
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