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Vol. 16, Issue 10, 4781-4791, October 2005
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* Chemistry and Chemical Biology Graduate Program, University of California, San Francisco, San Francisco, CA 94143;
Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94143
Submitted May 18, 2005;
Revised June 30, 2005;
Accepted July 6, 2005
Monitoring Editor: Thomas Fox
The mechanisms by which microorganisms sense and detoxify nitric oxide (.NO) are of particular interest due to the central role this molecule plays in innate immunity. We investigated the genetic basis of inducible nitric oxide (.NO) detoxification in Saccharomyces cerevisiae by characterizing the genome-wide transcriptional response to exogenously supplied .NO. Exposure to the .NO-generating compound dipropylenetriamine NONOate resulted in both a general stress response as well as a specific response characterized by the induction of a small set of genes, including the yeast flavohemoglobin YHB1, SSU1, and three additional uncharacterized open reading frames. Transcriptional induction of SSU1, which encodes a putative sulfite transporter, has previously been shown to require the zinc finger transcription factor Fzf1p. Deletion of Fzf1p eliminated the nitrosative stress-specific transcriptional response, whereas overexpression of Fzf1p recapitulated this response in the absence of exogenously supplied .NO. A cis-acting sequence unique to the promoter regions of Fzf1p-dependent genes was found to be sufficient to activate reporter gene activity in an .NO- and Fzf1p-dependent manner. Our results suggest that the presence of .NO or .NO derivatives activates Fzf1p leading to transcriptional induction of a discrete set of target genes that function to protect the cell from .NO-mediated stress.
Abbreviations used: APD, N-(3-aminopropyl)1,3-propane diamine; DPTA NONOate, dipropylenetriamine NONOate; .NO, nitric oxide; RNI, reactive nitrogen intermediate.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Address correspondence to: Joseph DeRisi (joe{at}derisilab.ucsf.edu)
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