Chen J, Pederson D S
Department of Microbiology and Molecular Genetics, University of Vermont, Burlington 05401.
J Biol Chem. 1993 Apr 5;268(10):7442-8.
Induction of heat shock genes is mediated by heat shock factor (HSF). Our recent genomic footprinting experiments demonstrate that HSF binds constitutively to perfect and imperfect heat shock elements (HSEs) in the HSP26 gene in yeast. Site-directed mutagenesis of the single perfect HSE, previously reported to not be involved in regulating gene expression, significantly reduces the rate of response of the gene to heat shock. However, the same mutation only slightly reduced the rate of accumulation of HSP26 mRNA during heat shock. Genomic footprinting experiments indicate that this lag in response to heat shock is due to the failure of HSF to bind efficiently to the mutated HSE. The rate of response to heat shock of synthetic promoters containing one, two, three, or seven perfect HSEs was similar to that observed for the wild-type HSP26 gene. These results suggest that the rate of response to heat shock is correlated with HSF occupancy of HSEs, rather than the number of HSEs in a promoter. As with the wild-type and mutant HSP26 genes, the rate of accumulation of mRNA from synthetic promoters increased only moderately with an increase in the number of HSEs. These results suggest that as few as two HSE-HSF complexes are sufficient to saturate HSF's target in the basal transcription apparatus.
热休克基因的诱导是由热休克因子(HSF)介导的。我们最近的基因组足迹实验表明,HSF在酵母中持续结合于HSP26基因中完美和不完美的热休克元件(HSE)。先前报道不参与调节基因表达的单个完美HSE的定点诱变,显著降低了该基因对热休克的反应速率。然而,相同的突变仅略微降低了热休克期间HSP26 mRNA的积累速率。基因组足迹实验表明,这种对热休克反应的滞后是由于HSF无法有效结合突变的HSE。含有一个、两个、三个或七个完美HSE的合成启动子对热休克的反应速率与野生型HSP26基因观察到的相似。这些结果表明,对热休克的反应速率与HSE的HSF占有率相关,而不是启动子中HSE的数量。与野生型和突变型HSP26基因一样,合成启动子的mRNA积累速率仅随着HSE数量的增加而适度增加。这些结果表明,少至两个HSE-HSF复合物就足以使基础转录装置中的HSF靶标饱和。