Heat shock-regulated transcription in vitro from a reconstituted chromatin template.
To investigate the mechanisms of transcriptional regulation of Drosophila heat shock genes we studied the activity of a heat shock promoter in vitro after reconstitution into chromatin. Increasing the duration of nucleosome assembly progressively inactivated a plasmid template when it was transcribed with extracts of either unshocked or heat-shocked Drosophila embryos, despite induction of the transcriptional activator heat shock factor. Addition of the general transcription factor IID (TFIID) before nucleosome assembly did not significantly relieve nucleosomal inhibition, but TFIID potentiated the promoter to be responsive to activation by heat shock factor in the heat shock transcription extract. The potentiation by TFIID could be related to the nucleosome-free, hypersensitive state of heat shock promoters previously observed in vivo before heat shock induction and may be necessitated by the need to expedite activation of heat shock genes in response to environmental stress.
Becker
P B
PB
Laboratory of Biochemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Rabindran
S K
SK
Wu
C
C
D000818:Animals; D002843:Chromatin / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000494:pharmacology; D004330:Drosophila / Q000196:embryology / Q000235:genetics*; D029721:Drosophila Proteins; D005786:Gene Expression Regulation; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000235:genetics*; D006358:Hot Temperature; D009707:Nucleosomes / Q000378:metabolism; D010957:Plasmids; D011401:Promoter Regions, Genetic; D035362:Transcription Factor TFIID; D014157:Transcription Factors / Q000494:pharmacology; D014158:Transcription, Genetic
D002843:Chromatin; D004268:DNA-Binding Proteins; D029721:Drosophila Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; C053134:Hsf protein, Drosophila; D009707:Nucleosomes; D035362:Transcription Factor TFIID; D014157:Transcription Factors