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

Genes Dev. 2002 Jan 1;16(1):101-13. doi: 10.1101/gad.949302.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Cell Cycle Proteins*
  • Chromosome Segregation / physiology
  • DNA-Binding Proteins
  • Fungal Proteins / physiology*
  • Kinetochores / physiology*
  • Molecular Sequence Data
  • Mutation
  • Saccharomyces cerevisiae / physiology*
  • Saccharomyces cerevisiae Proteins / physiology*
  • Schizosaccharomyces pombe Proteins*
  • Sequence Homology, Amino Acid

Substances

  • Cell Cycle Proteins
  • Ctf3 protein, S cerevisiae
  • DNA-Binding Proteins
  • Fungal Proteins
  • MCM16 protein, S cerevisiae
  • MCM22 protein, S cerevisiae
  • Mis6 protein, S pombe
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins