Hydrogen peroxide activates immediate binding of a Drosophila factor to DNA heat-shock regulatory element in vivo and in vitro.
The synthesis of heat-shock proteins via activation of heat-shock genes occurs in response to heat and various physical or chemical stressing agents. Transcriptional activation of heat-shock genes requires a heat-shock regulatory element in their promoter, to which a heat-shock specific transcription factor binds. In Drosophila cells, the heat-shock factor already exists in unstressed cells in an inactive form and acquires the capacity to bind to the heat-shock element following stress. The mechanism of this activation is not known: neither is it known whether the different stressing agents induce the heat-shock response through a common mechanism. We previously proposed that many agents known to induce the heat-shock response (substances interfering with respiratory metabolism, agents reacting with sulphydryl groups, metals, recovery from anaerobiosis and ischemia) might act via accumulation of reactive oxygen species, i.e. superoxide ion or H2O2. We show here that H2O2, introduced either in Drosophila cell cultures or in cell extracts, was able to activate heat-shock-element binding. Activation was rapid and H2O2 concentration dependent, with a threshold of 1 microM. These results were confirmed with mouse fibroblast cells. This very rapid activation, in vivo or in vitro, suggests a direct effect of H2O2 either on the heat-shock factor itself or on its activator.
Becker
J
J
Unité Associée au Centre National de la Recherche Scientifique 1135, Université Pierre et Marie Curie, Paris, France.
Mezger
V
V
Courgeon
A M
AM
Best-Belpomme
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000187:drug effects; D002460:Cell Line; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004330:Drosophila / Q000235:genetics / Q000378:metabolism*; D005809:Genes, Regulator / Q000187:drug effects*; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D014158:Transcription, Genetic / Q000187:drug effects*
D004268:DNA-Binding Proteins; D006360:Heat-Shock Proteins; D004247:DNA; D006861:Hydrogen Peroxide