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The Cdc2 protein kinase controls Cdc10/Sct1 complex formation

T Connolly1, M Caligiuri, D Beach

  • 1Howard Hughes Medical Institute, Cold Spring Harbor Laboratory, New York 11724, USA.

Molecular Biology of the Cell
|June 1, 1997
PubMed
Summary
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Phosphorylation of Cdc10 at serine-196 is crucial for its function in fission yeast cell cycle Start. This modification by Cdc2 kinase regulates the formation of the Cdc10/Sct1 transcription complex, controlling cell cycle progression.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Yeast Genetics

Background:

  • The Start transition in fission yeast (Schizosaccharomyces pombe) is a critical cell cycle checkpoint regulated by Cdc2 kinase and the Cdc10/Sct1 transcription complex.
  • Loss of function in these components results in G1 arrest and potential activation of the mating pathway.

Purpose of the Study:

  • To elucidate the molecular mechanisms regulating the Start transition in fission yeast.
  • To investigate the role of Cdc10 phosphorylation in the formation and function of the Cdc10/Sct1 transcription complex.

Main Methods:

  • Genetic analysis of fission yeast mutants.
  • Biochemical assays to assess protein-protein interactions and DNA-binding activity.
  • In vivo phosphorylation studies of Cdc10.

Related Experiment Videos

Main Results:

  • Serine-196 of Cdc10 is phosphorylated in vivo, and this phosphorylation is essential for its function.
  • Substitution of serine-196 with alanine (Cdc10 S196A) inactivates Cdc10 and prevents its association with Sct1.
  • Substitution with aspartic acid (Cdc10 S196D) restores DNA-binding and complex formation, alleviating the requirement for Cdc2 activity.
  • Cdc2 kinase activity is necessary for heteromeric Sct1/Cdc10 complex formation.

Conclusions:

  • Phosphorylation of Cdc10 at serine-196 by Cdc2 kinase is a key regulatory mechanism controlling the formation of the Sct1/Cdc10 transcription complex.
  • This phosphorylation event is essential for the regulation of the Start transition in the fission yeast cell cycle.