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Cdc28 activates exit from mitosis in budding yeast.

A D Rudner1, K G Hardwick, A W Murray

  • 1Department of Physiology, University of California, San Francisco, California 94143-0444, USA.

The Journal of Cell Biology
|June 28, 2000
PubMed
Summary

Budding yeast Cdc28 kinase activity is crucial for cell cycle progression. A CDC28-VF mutant delays mitosis exit by impairing the anaphase-promoting complex (APC) and Cdc20 association.

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Area of Science:

  • Cell Biology
  • Molecular Biology
  • Yeast Genetics

Background:

  • Cyclin-dependent kinase 1 (Cdk1), known as Cdc28 in budding yeast, regulates cell cycle transitions.
  • Inhibitory phosphorylation of Cdc28 normally facilitates the exit from mitosis.
  • Mutants lacking these phosphorylation sites can exhibit cell cycle delays.

Purpose of the Study:

  • To investigate the cell cycle role of the CDC28-T18V, Y19F (CDC28-VF) mutant, which lacks inhibitory phosphorylation sites.
  • To elucidate the mechanism behind the observed mitotic exit delay and hypersensitivity to mitotic arrest in CDC28-VF mutants.
  • To determine the impact of CDC28-VF on the anaphase-promoting complex (APC) activity and its regulators.

Main Methods:

  • Genetic manipulation to create the CDC28-VF mutant in budding yeast.
  • Cell cycle analysis to assess mitotic progression and exit.
  • Biochemical assays to measure APC activity and protein-protein interactions (Cdc20-APC complex).

Main Results:

  • The CDC28-VF mutant shows delayed exit from mitosis and increased sensitivity to mitotic arrest.
  • This phenotype is not due to altered inhibitory phosphorylation or kinase activity but reduced anaphase-promoting complex (APC) activity.
  • Specifically, Cdc20-dependent APC activity during mitosis is reduced in CDC28-VF, correlating with decreased Cdc20 association with the APC.
  • Hct1-dependent APC activity in G1 remains normal.

Conclusions:

  • Cdc28 kinase activity is essential for initiating the metaphase-to-anaphase transition and the subsequent anaphase-to-G1 transition.
  • The CDC28-VF mutant's defects highlight the importance of regulated Cdc28 activity for proper APC function, particularly Cdc20-mediated activation.
  • Proper regulation of APC activity by Cdc28 is critical for timely cell cycle progression in budding yeast.

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