Kirk N, Piper P W
Department of Biochemistry and Molecular Biology, University College London, U.K.
Yeast. 1991 Aug-Sep;7(6):539-46. doi: 10.1002/yea.320070602.
Heat-shock induction of heat-shock protein genes is due to a specific promoter element (the heat-shock element, HSE). This study used lacZ under HSE control (HSE-lacZ) to characterize HSE activity in Saccharomyces cerevisiae cells of different physiological states and differing genetic backgrounds. In batch fermentations HSE-lacZ induction by heat shock was maximal in exponential growth, and showed marked decline with the approach to stationary phase. Expression in the absence of heat shock was unaffected by growth phase, indicating that the growth-dependent expression of many yeast heat-shock genes uses promoter elements in addition to the HSE. Heat-induced expression was strongly influenced by the temperature at which cultures were grown. While basal, uninduced expression was constant during growth at different temperatures to 30 degrees C, induction by transfer to 39 degrees C was reduced by increases in growth temperature as low as 18-24 degrees C. Maximal HSE-lacZ induction (30- to 50-fold) was in cultures grown at low temperatures (18-24 degrees C), then heat shocked at 39 degrees C. Ethanol was a poor inducer. Mutations having little effect on HSE-lacZ expression included a respiratory petite; ubi4 (which inactivates the poly-ubiquitin gene); also ubc4 and ubc5 (which each inactivate one of the ubiquitin ligases involved in degradation of aberrant protein). pep4-3 increased both basal and induced beta-galactosidase about two-fold, probably because of slower turnover of this enzyme in pep4-3 strains.
热休克蛋白基因的热休克诱导是由于一个特定的启动子元件(热休克元件,HSE)。本研究使用在HSE控制下的lacZ(HSE-lacZ)来表征不同生理状态和不同遗传背景的酿酒酵母细胞中的HSE活性。在分批发酵中,热休克对HSE-lacZ的诱导在指数生长期最大,并随着接近稳定期而显著下降。在无热休克情况下的表达不受生长阶段的影响,这表明许多酵母热休克基因的生长依赖性表达除了HSE外还使用启动子元件。热诱导表达受到培养物生长温度的强烈影响。虽然在不同温度下生长至30℃期间基础的、未诱导的表达是恒定的,但转移至39℃时的诱导会因生长温度低至18-24℃的升高而降低。最大的HSE-lacZ诱导(30至50倍)出现在低温(18-24℃)下生长然后在39℃进行热休克的培养物中。乙醇是一种较差的诱导剂。对HSE-lacZ表达影响较小的突变包括呼吸缺陷型;ubi4(使多聚泛素基因失活);还有ubc4和ubc5(它们各自使参与异常蛋白质降解的一种泛素连接酶失活)。pep4-3使基础的和诱导的β-半乳糖苷酶都增加了约两倍,可能是因为在pep4-3菌株中这种酶的周转较慢。