Induction of sequence-specific binding of Drosophila heat shock activator protein without protein synthesis.
Drosophila tissue culture cells stimulated by heat shock contain high levels of heat shock activator protein, which binds specifically to the heat-shock control DNA element. In contrast, nonshocked cells have low basal levels of binding activity. Here, we show that within 30 seconds of heat shock of intact cells the sequence-specific binding activity in whole cell extracts increases significantly, reaching a plateau by 5 min after the start of the shock; removal of the heat stimulus returns the activity to basal levels. Known chemical inducers of heat-shock genes elicit a similar pattern of specific binding activity. Moreover, this pattern is observed in the presence of protein synthesis inhibitors, even if the stimulus-withdrawal is repeated sequentially through five cycles. Our results are inconsistent with models which propose proteolysis as the chief means of mediating heat-shock transcriptional control. Rather, they suggest that heat shock activator pre-exists in normal cells in a nonbinding form, which is converted upon cell stimulus to a high affinity, sequence-specific binding form, most probably by a post-translational modification. This conversion may be crucial for the transcriptional activation of heat shock genes.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins; D004140:Dinitrophenols / Q000494:pharmacology; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D029721:Drosophila Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D006358:Hot Temperature; D007700:Kinetics; D011506:Proteins / Q000378:metabolism*; D012980:Sodium Salicylate / Q000494:pharmacology; D014157:Transcription Factors; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D004140:Dinitrophenols; D029721:Drosophila Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; C053134:Hsf protein, Drosophila; D011506:Proteins; D014157:Transcription Factors; D004247:DNA; D012980:Sodium Salicylate