Viegas C A, Sebastião P B, Nunes A G, Sá-Correia I
Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, Lisbon, Portugal.
Appl Environ Microbiol. 1995 May;61(5):1904-9. doi: 10.1128/aem.61.5.1904-1909.1995.
During exponential growth at temperatures of 30 to 39 degrees C, the specific activity of H(+)-ATPase in the plasma membrane of Saccharomyces cerevisiae (assayed at the standard temperature 30 degrees C) increased with increases in growth temperature. In addition, the optimal temperature for in vitro activity of this ATPase was 42 degrees C. Therefore, the maximum values of ATPase activity were expected to occur in cells that grew within the supraoptimal range of temperatures. Activation induced by supraoptimal temperatures was not the result of increased synthesis of this membrane enzyme. When the growth temperature increased from 30 to 40 degrees C, expression of the essential PMA1 gene, monitored either by the level of PMA1 mRNA or the beta-galactosidase activity of the lacZ-PMA1 fusion, was reduced. Consistently, quantitative immunoassays showed that the ATPase content in the plasma membrane decreased. Like ATPase activity, the efficiency of the PMA2 promoter increased with increases in growth temperature in cells that had been grown at 30 to 39 degrees C, but its level of expression was several hundred-fold lower than that of PMA1. These results suggest that the major PMA1 ATPase is activated at supraoptimal temperatures.
在30至39摄氏度的指数生长期间,酿酒酵母质膜中H(+)-ATP酶的比活性(在标准温度30摄氏度下测定)随生长温度的升高而增加。此外,该ATP酶的体外活性最佳温度为42摄氏度。因此,预计ATP酶活性的最大值会出现在超最适温度范围内生长的细胞中。超最适温度诱导的激活不是这种膜酶合成增加的结果。当生长温度从30摄氏度升至40摄氏度时,通过PMA1 mRNA水平或lacZ-PMA1融合体的β-半乳糖苷酶活性监测的必需PMA1基因的表达降低。一致地,定量免疫测定表明质膜中的ATP酶含量下降。与ATP酶活性一样,在30至39摄氏度下生长的细胞中,PMA2启动子的效率随生长温度的升高而增加,但其表达水平比PMA1低数百倍。这些结果表明,主要的PMA1 ATP酶在超最适温度下被激活。