Regulation of Saccharomyces cerevisiae kinetochores by the type 1 phosphatase Glc7p

Ingrid Sassoon, Fedor F. Severin, Paul D. Andrews, Maria Rita Taba, Ken B. Kaplan, Anthony J. Ashford, Michael J R Stark, Peter K. Sorger, Anthony A. Hyman

Research output: Contribution to journalArticlepeer-review

124 Scopus citations

Abstract

We have investigated the role of protein phosphorylation in regulation of Saccharomyces cerevisiae kinetochores. By use of phosphatase inhibitors and a type I protein phosphatase mutant (glc7-10), we show that the microtubule binding activity, but not the centromeric DNA-binding activity, of the kinetochore complex is regulated by a balance between a protein kinase and the type 1 protein phosphatase (PP1) encoded by the GLC7 gene. glc7-10 mutant cells exhibit low kinetochore-microtubule binding activity in vitro and a high frequency of chromosome loss in vivo. Specifically, the Ndc10p component of the centromere DNA-binding CBF3 complex is altered by the glc7- 10 mutation; Ndc10p is hyperphosphorylated in glc7-10 extracts. Furthermore, addition of recombinant Ndc10p reconstitutes the microtubule-binding activity of a glc7-10 extract to wild-type levels. Finally, the glc7-10-induced mitotic arrest is abolished in spindle checkpoint mutants, suggesting that defects in kinetochore-microtubule interactions caused by hyperphosphorylation of kinetochore proteins activate the spindle checkpoint.

Original languageEnglish (US)
Pages (from-to)545-555
Number of pages11
JournalGenes and Development
Volume13
Issue number5
StatePublished - Mar 1 1999
Externally publishedYes

Keywords

  • Checkpoint
  • Kinetochore
  • Mitosis
  • Type 1 protein phosphatase

ASJC Scopus subject areas

  • Genetics
  • Developmental Biology

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