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首页> 外文期刊>Journal of Cellular Physiology >Effect of an alkaline shift on induction of the heat shock response in human fibroblasts.
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Effect of an alkaline shift on induction of the heat shock response in human fibroblasts.

机译：碱性的影响转变的感应人类成纤维细胞热休克反应。

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The simultaneous exposure of WI-38 human fibroblasts (HF) to a heat shock (45 degrees C, 30 min) and an alkaline shift (> or = pH 8.0) in the incubation medium increased and extended the expression of heat shock proteins (hsps). Hsp70 was the most prominent inducible hsp synthesized during the recovery phase after the double shock, and the increase in synthesis depended on the degree of alkalinization during the heat shock. The accumulation of inducible hsp70, which was shown by Western blotting to occur in the late part of the recovery period, was more pronounced in the cells exposed to alkaline medium during the heat shock. Northern blotting did not reveal any increase in hsp70 mRNA, although time course studies following the double shock indicated a more prolonged presence of mRNA. Hsp70 gene activation was evaluated by a gel retardation assay using a 32P-labelled DNA oligonucleotide containing the heat shock consensus element (HSE) and a heat shock-induced specific binding protein (heat shock transcription factor, HSTF) from the cell extract. Heat shock activated HSTF-DNA binding and induced hsp70 mRNA expression as well as the synthesis and accumulation of hsp70. Alkaline shift, which by itself did not induce hsps expression, activated HSTF DNA-binding. However, in combination with heat shock, alkaline shift enhanced and prolonged HSTF-HSE complex association and hsp expression at both mRNA and protein levels. Since the alkaline shift-induced activation of hsp gene does not allow full transcription, these results provide further support for the multistep nature of the heat shock transcriptional response.
机译：同时接触WI-38人类成纤维细胞(高频)热休克(45摄氏度,30分钟)和碱性pH值(> = 8.0)的转变孵化中增加和扩展热休克蛋白的表达(休克)。是最突出的诱导hsp合成在经济复苏阶段的双重打击后,和合成的增加取决于在热休克碱化度。诱导hsp70的积累,这是通过免疫印迹显示发生在晚了恢复期的一部分,更明显在细胞暴露于碱性介质中热休克。增加hsp70 mRNA,尽管时间进程研究的双重打击后表示更长期的信使rna的存在。激活是由凝胶阻滞评估分析使用32 p-labelled DNA寡核苷酸包含热休克共识元素(HSE)和一个热触觉具体结合蛋白(热休克转录因子,HSTF)细胞中提取。绑定和诱导hsp70 mRNA的表达hsp70的合成和积累。碱性转变,这本身并不引起休克蛋白表达,激活HSTF dna结合蛋白。然而,结合热休克,碱性增强和延长HSTF-HSE复杂转变协会和mRNA和hsp表达蛋白质的水平。hsp基因的激活不允许这些结果提供进一步的转录支持热火多阶段的特点休克转录反应。

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