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Eukaryotic Cell, August 2007, p. 1474-1485, Vol. 6, No. 8
1535-9778/07/$08.00+0     doi:10.1128/EC.00068-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Activation of the ADE Genes Requires the Chromatin Remodeling Complexes SAGA and SWI/SNF{triangledown}

Rebecca N. Koehler,{dagger} Nicole Rachfall,{ddagger} and Ronda J. Rolfes*

Department of Biology, Georgetown University, Washington, DC

Received 5 March 2007/ Accepted 3 June 2007

The activation of the ADE regulon genes requires the pair of transcription factors Bas1 and Pho2. In a genome-wide screen for additional regulators of the pathway, strains with mutations in multiple subunits of the chromatin remodeling complexes SAGA and SWI/SNF were uncovered. These mutants exhibited decreased expression of an ADE5,7-lacZ reporter and native ADE compared to the wild-type strains, but the expression of the BAS1 and PHO2 genes was not substantially decreased. An unregulated Bas1-Pho2 fusion protein depended upon SAGA and SWI/SNF activity to promote transcription of a reporter. A significant but low-level association of Gcn5-myc and Snf2-myc with the ADE5,7 promoter was independent of adenine growth conditions and independent of the presence of the activator proteins Bas1 and Pho2. However, the increase in occupancy of Bas1 and Pho2 at ADE5,7 depended on both SAGA and SWI/SNF. The loss of catalytic activity of both SAGA and SWI/SNF complexes in the gcn5{Delta} snf2{Delta} double mutant was severely detrimental to ADE-lacZ reporter expression and native ADE gene expression, indicating complementary roles for these complexes. We conclude that Bas1 and Pho2 do not recruit the SAGA and SWI/SNF complexes to the ADE5,7 promoter but that the remodeling complexes are necessary to increase the binding of Bas1 and Pho2 in response to the adenine regulatory signal. Our data support the model that the SAGA and SWI/SNF complexes engage in global surveillance that is necessary for the specific response by Bas1 and Pho2.

* Corresponding author. Mailing address: Department of Biology, Reiss Science Building 406, Georgetown University, Washington, DC 20057-1229. Phone: (202) 687-5881. Fax: (202) 687-5662. E-mail: rolfesr{at}georgetown.edu

{triangledown} Published ahead of print on 15 June 2007.

{dagger} Present address: US Military HIV Research Program, 1600 Gude Drive, Rockville, MD 20850.

{ddagger} Present address: Georg-August Universitaet, Grisebachstr. 8, Mikrobiologie und Genetik, Lab 110, Göttingen, Germany 37077.

Eukaryotic Cell, August 2007, p. 1474-1485, Vol. 6, No. 8
1535-9778/07/$08.00+0     doi:10.1128/EC.00068-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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