Monteiro G A, Supply P, Goffeau A, Sá-Correia I
Laboratório de Engenharia Bioquímica, Instituto Superior Técnico, Lisboa, Portugal.
Yeast. 1994 Nov;10(11):1439-46. doi: 10.1002/yea.320101107.
The expression of the PMA1 and PMA2 genes during Saccharomyces cerevisiae growth in medium with glucose plus increasing concentrations of ethanol was monitored by using PMA1-lacZ and PMA2-lacZ fusions and Northern blot hybridizations of total RNA probed with PMA1 gene. The presence of sub-lethal concentrations of ethanol enhanced the expression of PMA2 whereas it reduced the expression of PMA1. The inhibition of PMA1 expression by ethanol corresponded to a decrease in the content of plasma membrane ATPase as quantified by immunoassays. Although an apparent correspondence could exist between the increase of plasma membrane ATPase activity and the level of PMA2 expression, the maximal level of PMA2 expression remained about 200 times lower than PMA1. On the other hand, ethanol activated the plasma membrane H(+)-ATPase activity from a strain expressing only the PMA1 ATPase but did not activate that from a strain expressing only the PMA2 ATPase. These results provide evidence that in the presence of ethanol it is the PMA1 ATPase which is activated, probably by a post-translational mechanism and that the PMA2 ATPase is not involved.
利用PMA1-lacZ和PMA2-lacZ融合基因以及用PMA1基因探测的总RNA的Northern印迹杂交技术,监测酿酒酵母在含葡萄糖且乙醇浓度不断增加的培养基中生长时PMA1和PMA2基因的表达情况。亚致死浓度的乙醇存在时会增强PMA2的表达,而降低PMA1的表达。乙醇对PMA1表达的抑制作用与免疫分析定量的质膜ATP酶含量的降低相对应。尽管质膜ATP酶活性的增加与PMA2表达水平之间可能存在明显的对应关系,但PMA2表达的最大水平仍比PMA1低约200倍。另一方面,乙醇激活了仅表达PMA1 ATP酶的菌株的质膜H(+)-ATP酶活性,但未激活仅表达PMA2 ATP酶的菌株的该酶活性。这些结果证明,在乙醇存在的情况下,被激活的是PMA1 ATP酶,可能是通过翻译后机制激活的,而PMA2 ATP酶未参与其中。