Chromatin remodeling by GAGA factor and heat shock factor at the hypersensitive Drosophila hsp26 promoter in vitro.
The chromatin structure at the Drosophila hsp26 promoter in vivo is characterized by two DNase I-hypersensitive (DH) sites harboring regulatory elements. Proximal and distal DH sites are separated by a positioned nucleosome. To study the contribution of transcription factors to the establishment of this specific chromatin configuration we assembled nucleosomes on the hsp26 promoter using a cell-free reconstitution system derived from fly embryos. Both DH sites were readily reconstituted from extract components. They were separated by a nucleosome which was less strictly positioned than its in vivo counterpart. The interactions of GAGA factor and heat shock factor with their binding sites in chromatin occurred in two modes. Their interaction with binding sites in the nucleosome-free regions did not require ATP. In the presence of ATP both factors interacted also with nucleosomal binding sites, causing nucleosome rearrangements and a refinement of nucleosome positions. While chromatin remodeling upon transcription factor interaction has previously been interpreted to involve nucleosome disruption, the data suggest energy-dependent nucleosome sliding as main principle of chromatin reorganization.
Wall
G
G
European Molecular Biology Laboratory, Gene Expression Programme, Heidelberg, Germany.
Varga-Weisz
P D
PD
Sandaltzopoulos
R
R
Becker
P B
PB
D000818:Animals; D002843:Chromatin / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins; D004734:Energy Metabolism; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000235:genetics*; D018398:Homeodomain Proteins; D008957:Models, Genetic; D009068:Movement; D009707:Nucleosomes / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
D002843:Chromatin; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D018398:Homeodomain Proteins; C053134:Hsf protein, Drosophila; C087442:Hsp26 protein, Drosophila; D009707:Nucleosomes; D011994:Recombinant Proteins; D014157:Transcription Factors; C063460:Trl protein, Drosophila