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Molecular and Cellular Biology, December 1999, p. 8302-8313, Vol. 19, No. 12
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Yap1p Activates Gene Transcription in an Oxidant-Specific Fashion

Sean T. Coleman, Eric A. Epping, Susanne M. Steggerda,dagger and W. Scott Moye-Rowley*

Department of Physiology and Biophysics, University of Iowa, Iowa City, Iowa 52242

Received 14 April 1999/Returned for modification 25 May 1999/Accepted 25 August 1999

Positive regulation of gene expression by the yeast Saccharomyces cerevisiae transcription factor Yap1p is required for normal tolerance of oxidative stress elicited by the redox-active agents diamide and H2O2. Several groups have provided evidence that a cluster of cysteine residues in the extreme C terminus of the factor are required for normal modulation of Yap1p by oxidant challenge. Deletion of this C-terminal cysteine-rich domain (c-CRD) produces a protein that is highly active under both stressed and nonstressed conditions and is constitutively located in the nucleus. We have found that a variety of different c-CRD mutant proteins are hyperactive in terms of their ability to confer diamide tolerance to cells but fail to provide even normal levels of H2O2 resistance. Although the c-CRD mutant forms of Yap1p activate an artificial Yap1p-responsive gene to the same high level in the presence of either diamide or H2O2, these mutant factors confer hyperresistance to diamide but hypersensitivity to H2O2. To address this discrepancy, we have examined the ability of c-CRD mutant forms of Yap1p to activate expression of an authentic target gene required for H2O2 tolerance, TRX2. When assayed in the presence of c-CRD mutant forms of Yap1p, a TRX2-lacZ fusion gene fails to induce in response to H2O2. We have also identified a second cysteine-rich domain, in the N terminus (n-CRD), that is required for H2O2 but not diamide resistance and influences the localization of the protein. These data are consistent with the idea that the function of Yap1p is different at promoters of loci involved in H2O2 tolerance from promoters of genes involved in diamide resistance.

* Corresponding author. Mailing address: Department of Physiology and Biophysics, University of Iowa, Iowa City, IA 52242. Phone: (319)-335-7874. Fax: (319)-335-7330. E-mail: moyerowl{at}blue.weeg.uiowa.edu.

dagger Present address: Department of Biochemistry, Health Sciences Center, University of Virginia, Charlottesville, VA 22908.

Molecular and Cellular Biology, December 1999, p. 8302-8313, Vol. 19, No. 12
0270-7306/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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