Seppä Laura, Hänninen Anna-Liisa, Makarow Marja
Program in Cellular Biotechnology, Institute of Biotechnology, University of Helsinki, Viikinkaari 9, 00710 Helsinki, Finland.
Mol Microbiol. 2004 Apr;52(1):217-25. doi: 10.1111/j.1365-2958.2003.03959.x.
Thermal insult at 50 degrees C causes protein denaturation in yeast, but the cells survive if preconditioned at 37 degrees C. Survival depends on refolding of heat-denatured proteins. Refolding of cytoplasmic proteins requires Hsp104, the expression of which increases several-fold upon shift of the cells from physiological temperature 24 degrees C to 37 degrees C. We describe here a novel type of regulation of Hsp104, designated delayed upregulation (DUR). When Saccharomyces cerevisiae cells grown at 24 degrees C, preconditioned at 37 degrees C and treated briefly at 50 degrees C were shifted back to 24 degrees C, Hsp104 expression was negligible for 1 h, but increased then to a three to nine times higher level than that detected after growth at 24 degrees C, returning to normal after 5 h. A heat shock element (HSE) of the upstream sequence of HSP104 was necessary and sufficient for DUR, whereas stress response elements (STRE) were dispensable. Destruction of HSE plus all three STREs abolished Hsp104 expression, resulting in cell death after thermal insult. Deletion of MSN2/4, encoding transcription factors driving STRE-dependent gene expression, decreased DUR. Deletion of HOG1, encoding a heat-responsive and osmosensitive mitogen-activated protein kinase implicated to be functionally connected to Msn2/4p, abolished DUR. We suggest that DUR was regulated via HSE, required Hog1p and involved Msn2/4p-regulated gene products.
50摄氏度的热损伤会导致酵母中的蛋白质变性,但如果在37摄氏度下进行预处理,细胞就能存活。存活取决于热变性蛋白质的重折叠。细胞质蛋白质的重折叠需要Hsp104,当细胞从生理温度24摄氏度转移到37摄氏度时,Hsp104的表达会增加几倍。我们在此描述了一种新型的Hsp104调控方式,称为延迟上调(DUR)。当在24摄氏度下生长、在37摄氏度下预处理并在50摄氏度下短暂处理的酿酒酵母细胞转移回24摄氏度时,Hsp104的表达在1小时内可忽略不计,但随后会增加到比在24摄氏度下生长后检测到的水平高3至9倍,5小时后恢复正常。HSP104上游序列的热休克元件(HSE)对于DUR是必需且充分的,而应激反应元件(STRE)则是可有可无的。破坏HSE加上所有三个STRE会消除Hsp104的表达,导致热损伤后细胞死亡。缺失编码驱动STRE依赖性基因表达的转录因子的MSN2/4会降低DUR。缺失编码与Msn2/4p功能相关的热响应和渗透敏感丝裂原活化蛋白激酶的HOG1会消除DUR。我们认为DUR是通过HSE调控的,需要Hog1p并涉及Msn2/4p调控的基因产物。