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Molecular and Cellular Biology, September 2002, p. 6148-6157, Vol. 22, No. 17
0270-7306/02/$04.00+0     DOI: 10.1128/MCB.22.17.6148-6157.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

GAGA Factor and the TFIID Complex Collaborate in Generating an Open Chromatin Structure at the Drosophila melanogaster hsp26 Promoter

Boris A. Leibovitch,1 Quinn Lu,1,{dagger} Lawrence R. Benjamin,2,{ddagger} Yingyun Liu,2 David S. Gilmour,2 and Sarah C. R. Elgin1*

Department of Biology, Washington University, St. Louis, Missouri 63130,1 Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 168022

Received 27 February 2002/ Returned for modification 2 May 2002/ Accepted 30 May 2002

The upstream regulatory region of the Drosophila melanogaster hsp26 gene includes two DNase I-hypersensitive sites (DH sites) that encompass the critical heat shock elements. This chromatin structure is required for heat shock-inducible expression and depends on two (CT)n•(GA)n elements bound by GAGA factor. To determine whether GAGA factor alone is sufficient to drive formation of the DH sites, we have created flies with an hsp26/lacZ transgene wherein the entire DNA segment known to interact with the TFIID complex has been replaced by a random sequence. The replacement results in a loss of heat shock-inducible hsp26 expression and drastically diminishes nuclease accessibility in the chromatin of the regulatory region. Chromatin immunoprecipitation experiments show that the decrease in TFIID binding does not reduce GAGA factor binding. In contrast, the loss of GAGA factor binding resulting from (CT)n mutations decreases TFIID binding. These data suggest that both GAGA factor and TFIID are necessary for formation of the appropriate chromatin structure at the hsp26 promoter and predict a regulatory mechanism in which GAGA factor binding precedes and contributes to the recruitment of TFIID.

* Corresponding author. Mailing address: Department of Biology, Washington University, CB1229, One Brookings Dr., St. Louis, MO 63130. Phone: (314) 935-5348. Fax: (314) 935-5125. E-mail: selgin{at}biology.wustl.edu.

{dagger} Present address: GlaxoSmithKline, King of Prussia, PA 19406.

{ddagger} Present address: National Cancer Institute, Bethesda, MD 20892.

Molecular and Cellular Biology, September 2002, p. 6148-6157, Vol. 22, No. 17
0022-538X/02/$04.00+0     DOI: 10.1128/MCB.22.17.6148-6157.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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