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Molecular and Cellular Biology, February 2008, p. 1326-1337, Vol. 28, No. 4
0270-7306/08/$08.00+0     doi:10.1128/MCB.01219-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Yap5 Is an Iron-Responsive Transcriptional Activator That Regulates Vacuolar Iron Storage in Yeast{triangledown}

Liangtao Li, Dustin Bagley, Diane M. Ward, and Jerry Kaplan*

Department of Pathology, School of Medicine, University of Utah, Salt Lake City, Utah 84132

Received 9 July 2007/ Returned for modification 20 July 2007/ Accepted 21 November 2007

The transporter Ccc1 imports iron into the vacuole, which is the major site of iron storage in fungi and plants. CCC1 mRNA is destabilized under low-iron conditions by the binding of Cth1 and Cth2 to the 3' untranslated region (S. Puig, E. Askeland, and D. J. Thiele, Cell 120:99-110, 2005). Here, we show that the transcription of CCC1 is stimulated by iron through a Yap consensus site in the CCC1 promoter. We identified YAP5 as being the iron-sensitive transcription factor and show that a yap5{Delta} strain is sensitive to high iron. Green fluorescent protein-tagged Yap5 is localized to the nucleus and occupies the CCC1 promoter independent of the iron concentration. Yap5 contains two cysteine-rich domains, and the mutation of the cysteines to alanines in each of the domains affects the transcription of CCC1 but not DNA binding. The fusion of the Yap5 cysteine-containing domains to a GAL4 DNA binding domain results in iron-sensitive GAL1-lacZ expression. Iron affects the sulfhydryl status of Yap5, which is indicative of the generation of intramolecular disulfide bonds. These results show that Yap5 is an iron-sensing transcription factor and that iron regulates transcriptional activation.

* Corresponding author. Mailing address: Department of Pathology, University of Utah School of Medicine, Salt Lake City, UT 84132. Phone: (801) 581-7427. Fax: (801) 585-6364. E-mail: jerry.kaplan{at}path.utah.edu

{triangledown} Published ahead of print on 10 December 2007.

Molecular and Cellular Biology, February 2008, p. 1326-1337, Vol. 28, No. 4
0270-7306/08/$08.00+0     doi:10.1128/MCB.01219-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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