Abstract

We observed that the transcription of Saccharomyces cerevisiae cytoplasmic thiol peroxidase type II (cTPx II) (YDR453C) is regulated in response to various stresses (e.g. oxidative stress, carbon starvation, and heat-shock). It has been suggested that both transcription-activating proteins, Yap1p and Skn7p, regulate the transcription of cTPx II upon exposure to oxidative stress. However, a dramatic loss of transcriptional response to various stresses in yeast mutant strains lacking both Msn2p and Msn4p suggests that the transcription factors act as a principal transcriptional activator. In addition to two Yap1p response elements (YREs), TTACTAA and TTAGTAA, the presence of two stress response elements (STREs) (CCCCT) in the upstream sequence of cTPx II also suggests that Msn2p/Msn4p could control stress-induced expression of cTPx II. Analysis of the transcriptional activity of site-directed mutagenesis of the putative STREs (STRE1 and STRE2) and YREs (TRE1 and YRE2) in terms of the activity of a lacZ reporter gene under control of the cTPx II promoter indicates that STRE2 acts as a principal binding element essential for transactivation of the cTPx II promoter. The transcriptional activity of the cTPx II promoter was exponentially increased after postdiauxic growth. The transcriptional activity of the cTPx II promoter is greatly increased by rapamycin. Deletion of Tor1, Tor2, Ras1, and Ras2 resulted in a considerable induction when compared with their parent strains, suggesting that the transcription of cTPx II is under negative control of the Ras/cAMP and target of rapamycin signaling pathways. Taken together, these results suggest that cTPx II is a target of Msn2p/Msn4p transcription factors under negative control of the Ras-protein kinase A and target of rapamycin signaling pathways. Furthermore, the accumulation of cTPx II upon exposure to oxidative stress and during the postdiauxic shift suggests an important antioxidant role in stationary phase yeast cells.

Highlights

  • Growing cells are continuously challenged by reactive oxygen species

  • The Transcription of cytoplasmic thiol peroxidase type II (cTPx II) Is Induced in Response to Oxidative Stress—Previously, we reported that exposure of yeast cells harboring the cTPx II-lacZ gene fusion vector to oxidative stress increases the ␤-galactosidase activities [22]

  • Comparative Western blot (Fig. 1B) and RT-PCR (Fig. 1C) analyses showed that the protein and mRNA levels in early log-phase cells were very low but that the expression of cTPx II was dramatically induced in response to exposure of the yeasts to H2O2 and diamide

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Summary

Introduction

Growing cells are continuously challenged by reactive oxygen species. Reactive oxygen species are potent oxidants capable of damaging all cellular components including DNA, protein, and membrane lipid. The Transcription of cTPx II Is Induced in Response to Oxidative Stress—Previously, we reported that exposure of yeast cells harboring the cTPx II-lacZ gene fusion vector to oxidative stress increases the ␤-galactosidase activities [22].

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