Sgt1 dimerization is required for yeast kinetochore assembly

Parmil K. Bansal, Amanda Nourse, Rashid Abdulle, Katsumi Kitagawa

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The kinetochore, which consists of DNA sequence elements and structural proteins, is essential for high-fidelity chromosome transmission during cell division. In budding yeast, Sgt1 and Hsp90 help assemble the core kinetochore complex CBF3 by activating the CBF3 components Skp1 and Ctf13. In this study, we show that Sgt1 forms homodimers by performing in vitro and in vivo immunoprecipitation and analytical ultracentrifugation analyses. Analyses of the dimerization of Sgt1 deletion proteins showed that the Skp1-binding domain (amino acids 1-211) contains the Sgt1 homodimerization domain. Also, the Sgt1 mutant proteins that were unable to dimerize also did not bind Skp1, suggesting that Sgt1 dimerization is important for Sgt1-Skp1 binding. Restoring dimerization activity of a dimerization-deficient sgt1 mutant (sgt1-L31P) by using the CENP-B (centromere protein-B) dimerization domain suppressed the temperature sensitivity, the benomyl sensitivity, and the chromosome missegregation phenotype of sgt1-L31P. These results strongly suggest that Sgt1 dimerization is required for kinetochore assembly.

Original languageEnglish (US)
Pages (from-to)3586-3592
Number of pages7
JournalJournal of Biological Chemistry
Volume284
Issue number6
DOIs
StatePublished - Feb 6 2009
Externally publishedYes

Fingerprint

Kinetochores
Dimerization
Yeast
Yeasts
Centromere Protein B
Chromosomes
S-Phase Kinase-Associated Proteins
Protein Multimerization
Benomyl
Saccharomycetales
Ultracentrifugation
Mutant Proteins
Immunoprecipitation
Cell Division
DNA sequences
Phenotype
Amino Acids
Cells
Temperature
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sgt1 dimerization is required for yeast kinetochore assembly. / Bansal, Parmil K.; Nourse, Amanda; Abdulle, Rashid; Kitagawa, Katsumi.

In: Journal of Biological Chemistry, Vol. 284, No. 6, 06.02.2009, p. 3586-3592.

Research output: Contribution to journalArticle

Bansal, Parmil K. ; Nourse, Amanda ; Abdulle, Rashid ; Kitagawa, Katsumi. / Sgt1 dimerization is required for yeast kinetochore assembly. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 6. pp. 3586-3592.
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