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MBC in Press, published online ahead of print March 29, 2006
Mol. Biol. Cell 10.1091/mbc.E05-11-1093

A more recent version of this article appeared on June 1, 2006
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Submitted on December 1, 2005
Revised on February 13, 2006
Accepted on March 8, 2006

The Function and Regulation of Budding Yeast Swe1 in Response to Interrupted DNA Synthesis

Hong Liu* and Yanchang Wang*{dagger}

{dagger}Department of Biomedical Sciences, College of Medicine and *Department of Biology, Florida State University, Tallahassee, FL 32306

Monitoring Editor: Mark Solomon

Periodically regulated cyclin dependent kinase (Cdk) is required for DNA synthesis and mitosis. Hydroxyurea (HU) inhibits DNA synthesis by depleting dNTPs, the basic unit for DNA synthesis. HU treatment triggers the S-phase checkpoint, which arrests cells at S-phase, inhibits late origin firing, and stabilizes replication forks. Using budding yeast as a model system, we found that Swe1, a negative regulator of Cdk, appears at S-phase and accumulates in HU treatment cells. Interestingly, this accumulation is not dependent on S-phase checkpoint. {Delta}hsl1, {Delta}hsl7, and cdc5-2 mutants, which have defects in Swe1 degradation, show HU sensitivity because of high Swe1 protein levels. We further demonstrated that their HU sensitivity is not a result of DNA damage accumulation or incomplete DNA synthesis, instead the sensitivity is due to their dramatically delayed recovery from HU-induced S-phase arrest. Strikingly, our in vivo data indicate that Swe1 inhibits the kinase activity of Clb2-Cdk1, but not that of Clb5-Cdk1. Therefore, S-phase accumulated Swe1 prevents Clb2-Cdk1 mediated mitotic activities, but has little effects on Clb5-Cdk1 associated S-phase progression.

Address correspondence to: Yanchang Wang (yanchang.wang{at}med.fsu.edu)




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