In budding yeast, reactive oxygen species induce both RAS-dependent and RAS-independent cell cycle-specific arrest.
The role of mild oxidative stresses elicited by diethylmaleate (DEM)-induced glutathione depletion in the progression of the yeast cell cycle has been investigated. We found that different wild-type strains are sensitive to oxidative stresses induced by similar DEM doses: approximately 1 mM on YPD plates, 5-10 mM in shaken flasks. At lower doses, DEM caused a transient decrease in growth rate, largely because of a decreased G1-to-S transition. Treatment with higher DEM doses leads to complete growth arrest, with most cells found in the unbudded G1 phase of the cell cycle. DEM treatment resulted in transcriptional induction of stress-responsive element (STRE)-controlled genes and was relieved by treatment with the antioxidant N-acetyl cysteine. Reciprocal shift experiments with cdc25 and cdc28 mutants showed that the major cell cycle arrest point was located in the Start area, at or near the CDC25-mediated step, before the step mediated by the CDC28 cyclin-dependent kinase. The DEM-induced G1 arrest requires a properly regulated RAS pathway and can be bypassed by overexpressing the G1-specific cyclin CLN2. However, cells with either a deregulated RAS pathway or overexpressing CLN2 failed to grow and arrested as budded cells, indicating that a second DEM-sensitive cell cycle step exists.
Wanke
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Dipartimento di Biotecnologie e Bioscienze, Università degli Studi di Milano-Bicocca, Milan, Italy.
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D000111:Acetylcysteine / Q000494:pharmacology; D002453:Cell Cycle / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects / Q000235:genetics; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D016213:Cyclins / Q000235:genetics; D004247:DNA / Q000032:analysis; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D005800:Genes, Fungal; D011905:Genes, ras; D005978:Glutathione / Q000378:metabolism; D007399:Interphase / Q000235:genetics; D008298:Maleates / Q000494:pharmacology; D018384:Oxidative Stress / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D016213:Cyclins; D008298:Maleates; D017382:Reactive Oxygen Species; D004247:DNA; D018844:Cyclin-Dependent Kinases; C014476:diethyl maleate; D005978:Glutathione; D000111:Acetylcysteine