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Originally published as Genetics Published Articles Ahead of Print on January 16, 2005.

Genetics, Vol. 169, 1343-1352, March 2005, Copyright © 2005
doi:10.1534/genetics.104.037143

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The Heme Activator Protein Hap1 Represses Transcription by a Heme-Independent Mechanism in Saccharomyces cerevisiae

Thomas Hon*, Hee Chul Lee*, Zhanzhi Hu{dagger}, Vishwanath R. Iyer{dagger} and Li Zhang*,1

* Department of Environmental Health Sciences, Columbia University, Mailman School of Public Health, New York, New York 10032
{dagger} Institute for Cellular and Molecular Biology, University of Texas, Austin, Texas 78712

1 Corresponding author: Department of Environmental Health Sciences, Columbia University, Mailman School of Public Health, 60 Haven Ave., B-1, New York, NY 10032.
E-mail: lz2115{at}columbia.edu

The yeast heme activator protein Hap1 binds to DNA and activates transcription of genes encoding functions required for respiration and for controlling oxidative damage, in response to heme. Hap1 contains a DNA-binding domain with a C6 zinc cluster motif, a coiled-coil dimerization element, typical of the members of the yeast Gal4 family, and an acidic activation domain. The regulation of Hap1 transcription-activating activity is controlled by two classes of Hap1 elements, repression modules (RPM1–3) and heme-responsive motifs (HRM1–7). Previous indirect evidence indicates that Hap1 may repress transcription directly. Here we show, by promoter analysis, by chromatin immunoprecipitation, and by electrophoretic mobility shift assay, that Hap1 binds directly to DNA and represses transcription of its own gene by at least 20-fold. We found that Hap1 repression of the HAP1 gene occurs independently of heme concentrations. While DNA binding is required for transcriptional repression by Hap1, deletion of Hap1 activation domain and heme-regulatory elements has varying effects on repression. Further, we found that repression by Hap1 requires the function of Hsp70 (Ssa), but not Hsp90. These results show that Hap1 binds to its own promoter and represses transcription in a heme-independent but Hsp70-dependent manner.




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