title: Structural and functional dissection of Mif2p, a conserved DNA-binding kinetochore protein.
abstract: Mif2p is the budding-yeast orthologue of the mammalian centromere-binding protein CENP-C. We have mapped domains of Saccharomyces cerevisiae Mif2p and studied the phenotyptic consequences of their deletion. Using chromatin immunoprecipitation (ChIP) and electrophoretic mobility shift assays, we have further shown that Mif2p binds in the CDEIII region of the budding-yeast centromere, probably in close spatial association with Ndc10p. Moreover, ChIP experiments show that Mif2p recruits to yeast kinetochores a substantial subset of inner and outer kinetochore proteins, but not the Ndc80 or Spc105 complexes. We have determined the crystal structure of the C-terminal, dimerization domain of Mif2p. It has a "cupin" fold, extremely similar both in polypeptide chain conformation and in dimer geometry to the dimerization domain of a bacterial transcription factor. The Mif2p dimer seems to be part of an enhanceosome-like structure that nucleates kinetochore assembly in budding yeast.
authors.affiliation: Jack and Eileen Connors Structural Biology Laboratory, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: R L
authors.initials: RL
authors.lastname: Cohen
authors.forename: C W
authors.initials: CW
authors.lastname: Espelin
authors.forename: P
authors.initials: P
authors.lastname: De Wulf
authors.forename: P K
authors.initials: PK
authors.lastname: Sorger
authors.forename: S C
authors.initials: SC
authors.lastname: Harrison
authors.forename: K T
authors.initials: KT
authors.lastname: Simons
mesh_terms: D000595:Amino Acid Sequence; D018360:Crystallography, X-Ray / Q000379:methods; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000502:physiology; D019281:Dimerization; D015966:Gene Expression Regulation, Fungal; D018386:Kinetochores / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010641:Phenotype; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000502:physiology*; D004718:Saccharomycetales; D017386:Sequence Homology, Amino Acid
chemical_list: C081071:CBF2 protein, S cerevisiae; D004268:DNA-Binding Proteins; C083768:MIF2 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins