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Related Experiment Videos

Characterization of the checkpoint gene RAD53/MEC2 in Saccharomyces cerevisiae

S Kim1, T A Weinert

  • 1Department of Molecular and Cellular Biology, University of Arizona, Tucson 85721, USA.

Yeast (Chichester, England)
|June 30, 1997
PubMed
Summary

RAD53 is essential for cell viability and DNA replication checkpoints in Saccharomyces cerevisiae. Its protein kinase domain is crucial for both essential and checkpoint functions, with distinct regulatory elements.

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Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • Genetics

Background:

  • Saccharomyces cerevisiae cells with RAD53/MEC2 mutations exhibit defects in S/M and G2/M checkpoints.
  • RAD53 plays a critical role in cell cycle control and DNA damage response.

Purpose of the Study:

  • To investigate the essential and checkpoint functions of RAD53.
  • To elucidate the role of the RAD53 protein kinase domain in cell viability and cell cycle progression.

Main Methods:

  • Gene sequencing to identify RAD53.
  • Genetic analysis of null mutants and suppressors.
  • Site-specific mutagenesis to inactivate the protein kinase domain.
  • Overexpression studies of RAD53 and its kinase domain.

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Main Results:

  • RAD53 is identical to SPK1 and is essential for cell viability.
  • The protein kinase domain of RAD53 is critical for all tested functions.
  • Overexpression of RAD53 or its kinase domain causes cell-cycle delay independent of known checkpoint genes.
  • Complex regulatory elements were identified in the RAD53 promoter region.

Conclusions:

  • RAD53 has both essential cell viability and DNA checkpoint functions, which can be genetically separated.
  • The protein kinase activity of RAD53 is indispensable for its diverse cellular roles.
  • RAD53-mediated cell-cycle delay may operate downstream of or independently from canonical checkpoint pathways.