Prändl R, Hinderhofer K, Eggers-Schumacher G, Schöffl F
Lehrstuhl für Allgemeine Genetik, Eberhard-Karls-Universität Tübingen, Germany.
Mol Gen Genet. 1998 May;258(3):269-78. doi: 10.1007/s004380050731.
Organisms synthesize heat shock proteins (HSPs) in response to sublethal heat stress and concomitantly acquire increased tolerance against a subsequent, otherwise lethal, heat shock. Heat shock factor (HSF) is essential for the transcription of many HSP genes. We report the isolation of two HSF genes, HSF3 and HSF4, from an Arabidopsis cDNA library. Transgenic Arabidopsis plants were generated containing constructs that allow expression of HSF3 and HSF4 or the respective translational beta-glucuronidase (GUS) fusions. Overexpression of HSF3 or HSF3-GUS, but not of HSF4 or HSF4-GUS, causes HSP synthesis at the non-heat-shock temperature of 25 degrees C in transgenic Arabidopsis. In transgenic plants bearing HSF3/HSF3-GUS, transcription of several heat shock genes is derepressed. Electrophoretic mobility shift assays suggest that derepression of the heat shock response is mediated by HSF3/HSF3-GUS functioning as transcription factor. HSF3/HSF3-GUS-overexpressing Arabidopsis plants show an increase in basal thermotolerance, indicating the importance of HSFs and HSF-regulated genes as determinants of thermoprotective processes. Plants transgenic for HSF3/HSF3-GUS exhibit no other obvious phenotypic alterations. Derepression of HSF activity upon overexpression suggests the titration of a negative regulator of HSF3 or an intrinsic constitutive activity of HSF3. We assume that stable overexpression of HSFs may be applied to other organisms as a means of derepressing the heat shock response.
生物体在受到亚致死热胁迫时会合成热休克蛋白(HSPs),并随之获得对随后的、否则会致死的热休克的耐受性增强。热休克因子(HSF)对于许多HSP基因的转录至关重要。我们报道了从拟南芥cDNA文库中分离出两个HSF基因,即HSF3和HSF4。构建了允许表达HSF3和HSF4或各自的翻译β-葡萄糖醛酸酶(GUS)融合体的载体,并生成了转基因拟南芥植株。在转基因拟南芥中,HSF3或HSF3-GUS的过表达会导致在25℃的非热休克温度下合成HSP,而HSF4或HSF4-GUS的过表达则不会。在携带HSF3/HSF3-GUS的转基因植株中,几个热休克基因的转录被解除抑制。电泳迁移率变动分析表明,热休克反应的解除抑制是由作为转录因子发挥作用的HSF3/HSF3-GUS介导的。过表达HSF3/HSF3-GUS的拟南芥植株的基础耐热性增强,这表明HSFs和HSF调控的基因作为热保护过程的决定因素具有重要性。转HSF3/HSF3-GUS基因的植株没有表现出其他明显的表型改变。过表达时HSF活性的解除抑制表明对HSF3的负调节因子进行了滴定或HSF3具有内在的组成型活性。我们认为,HSFs的稳定过表达可作为一种解除热休克反应抑制的手段应用于其他生物体。