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PMID: 18701705 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Structural and functional dissection of Mif2p, a conserved DNA-binding kinetochore protein.

Molecular biology of the cell ·Vol. 19 ·No. 10 ·2008-10-00 ·Pages 4480-91

Cohen RL, Espelin CW, De Wulf P, Sorger PK, Harrison SC, Simons KT

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.

MeSH Terms
Amino Acid Sequence Crystallography, X-Ray/methods DNA-Binding Proteins/chemistry,physiology Dimerization Gene Expression Regulation, Fungal Kinetochores/chemistry,metabolism Molecular Sequence Data Phenotype Protein Binding Protein Structure, Tertiary Saccharomyces cerevisiae/metabolism Saccharomyces cerevisiae Proteins/chemistry,physiology Saccharomycetales Sequence Homology, Amino Acid
Chemicals
CBF2 protein, S cerevisiae DNA-Binding Proteins MIF2 protein, S cerevisiae Saccharomyces cerevisiae Proteins
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Cohen R L
Jack and Eileen Connors Structural Biology Laboratory, Harvard Medical School, Boston, MA 02115, USA.
Espelin C W
De Wulf P
Sorger P K
Harrison S C
Simons K T
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Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1939-4586
DOI
10.1091/mbc.e08-03-0297
Published
2008-10-00
Epub
2008-00-13
Pages
4480-91
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC2555937
Subset
IM
Grants
NCI NIH HHS · R01 CA084179 · United States
Howard Hughes Medical Institute · United States
NCI NIH HHS · CA-84179 · United States
NIGMS NIH HHS · R01 GM051464 · United States
NIGMS NIH HHS · GM-51464 · United States
External Links
PubMed source DOI: 10.1091/mbc.e08-03-0297 PMC Full Text View journal details
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