Coordinate changes in heat shock element-binding activity and HSP70 gene transcription rates in human cells.
Activation of human heat shock gene transcription by heat shock, heavy metal ions, and amino acid analogs required the heat shock element (HSE) in the HSP70 promoter. Both heat shock- and metal ion-induced HSP70 gene transcription occurred independently of protein synthesis, whereas induction by amino acid analogs required protein synthesis. We identified a HSE-binding activity from control cells which was easily distinguished by a gel mobility shift assay from the stress-induced HSE-binding activity which appeared following heat shock or chemically induced stress. The kinetics of HSP70 gene transcription paralleled the rapid appearance of stress-induced HSE-binding activity. During recovery from heat shock, both the rate of HSP70 gene transcription and stress-induced HSE-binding activity levels declined and the control HSE-binding activity reappeared. The DNA contacts of the control and stress-induced HSE-binding activities deduced by methylation interference were similar but not identical. While stable complexes with HSE were formed with extracts from both control and stressed cells in vitro at 25 degrees C, only the stress-induced complex was detected when binding reactions were performed at elevated temperatures.
Mosser
D D
DD
Department of Biochemistry, Molecular Biology, Northwestern University, Evanston, Illinois 60208.
Theodorakis
N G
NG
Morimoto
R I
RI
D001384:Azetidines / Q000494:pharmacology; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics*; D006358:Hot Temperature; D006801:Humans; D007700:Kinetics; D008670:Metals / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects
D001384:Azetidines; D006360:Heat-Shock Proteins; D008670:Metals; D011500:Protein Synthesis Inhibitors; D004247:DNA