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

R. L. Cohen, C. W. Espelin, P. De Wulf, P. K. Sorger, S. C. Harrison, K. T. Simons

Research output: Contribution to journalArticle

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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.

Original languageEnglish
Pages (from-to)4480-4491
Number of pages12
JournalMolecular Biology of the Cell
Volume19
Issue number10
DOIs
Publication statusPublished - Oct 2008

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Kinetochores
Saccharomycetales
DNA-Binding Proteins
Dissection
Centromere
Chromatin Immunoprecipitation
Dimerization
Electrophoretic Mobility Shift Assay
Protein C
Saccharomyces cerevisiae
Carrier Proteins
Transcription Factors
Yeasts
Peptides
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Cohen, R. L., Espelin, C. W., De Wulf, P., Sorger, P. K., Harrison, S. C., & Simons, K. T. (2008). Structural and functional dissection of Mif2p, a conserved DNA-binding kinetochore protein. Molecular Biology of the Cell, 19(10), 4480-4491. https://doi.org/10.1091/mbc.E08-03-0297

Structural and functional dissection of Mif2p, a conserved DNA-binding kinetochore protein. / Cohen, R. L.; Espelin, C. W.; De Wulf, P.; Sorger, P. K.; Harrison, S. C.; Simons, K. T.

In: Molecular Biology of the Cell, Vol. 19, No. 10, 10.2008, p. 4480-4491.

Research output: Contribution to journalArticle

Cohen, RL, Espelin, CW, De Wulf, P, Sorger, PK, Harrison, SC & Simons, KT 2008, 'Structural and functional dissection of Mif2p, a conserved DNA-binding kinetochore protein', Molecular Biology of the Cell, vol. 19, no. 10, pp. 4480-4491. https://doi.org/10.1091/mbc.E08-03-0297
Cohen, R. L. ; Espelin, C. W. ; De Wulf, P. ; Sorger, P. K. ; Harrison, S. C. ; Simons, K. T. / Structural and functional dissection of Mif2p, a conserved DNA-binding kinetochore protein. In: Molecular Biology of the Cell. 2008 ; Vol. 19, No. 10. pp. 4480-4491.
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