Ctf3p, the Mis6 budding yeast homolog, interacts with Mcm22p and Mcm16p at the yeast outer kinetochore

• Published in: Genes & development Vol. 16; no. 1; pp. 101 - 113
• Main Authors: Measday, Vivien, Hailey, Dale W, Pot, Isabelle, Givan, Scott A, Hyland, Katherine M, Cagney, Gerard, Fields, Stan, Davis, Trisha N, Hieter, Philip
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory Press 01.01.2002
• Subjects: Amino Acid Sequence; Cell Cycle Proteins; CEN gene; Chromosome Segregation - physiology; Ctf19 protein; Ctf3 protein; DNA-Binding Proteins; Fungal Proteins - physiology; Kinetochores - physiology; Mcm16 protein; Mcm22 protein; Mis6 protein; Molecular Sequence Data; Mutation; Research Paper; Saccharomyces cerevisiae; Saccharomyces cerevisiae - physiology; Saccharomyces cerevisiae Proteins - physiology; Schizosaccharomyces pombe; Schizosaccharomyces pombe Proteins; Sequence Homology, Amino Acid
• ISSN: 0890-9369; 1549-5477
• DOI: 10.1101/gad.949302

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, onto CEN 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.