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Nucleic Acids Research, 1995, Vol. 23, No. 13 2479-2487
© 1995

MOLECULAR BIOLOGY

Dual regulation of the Drosophilla hsp26 promoter in vitro

Raphael Sandaltzopolus, Catherine Mitchelmore+, Edgar Bonte, Gayl Wal and Peter B. Becker*

Gene Expression Programme, European Molecular Biology Meyerhoffstrasse 1,69117 Heidelburg, germany

*To whom corespondence should be addressed

Received March 22, 1995. Accepted May 18, 1995.

Efficient heat shock induction of Drosophila hsp26 gene transcription in vivo requires binding sites for heat shock factor (HSF) and GAGA factor (GAF) close to the TATA box (proximal elements) as well as 350 bp upstream of the start site of transcription (distal elements). We have evaluated the contribution of hsp26 promoter sequences to transciiptional activity in extracts from either heat shocked or unstressed fly embryos. Efficient transcription In either extract was governed by distinct regulatory principles. Transcription in extracts from unstressed embryos relied solely on GAGA elements which efficiently cocnteracted repression by abundant non-specific DNA-blndfng proteins. Transcription in extracts from heat shocked embryos depended only a little on GAGA elements, relying mainly on functional HSEs. Constitutively active recombinant HSF or native factor in an extract from heat shocked embryos was able to truly activate transcriptin essentially via proximal HSEs, but not when bound to distal sites. These two modes of regulation in vitro may correspond to the two functional states of the promoter before and after heat shock in vivo

+Present address: Department of Biochemistry C, The Panum Institute, Bledgamsvej,3, DK 2200 Cophegen N, Denmark


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