Van Aelst L, Boy-Marcotte E, Camonis J H, Thevelein J M, Jacquet M
Laboratorium voor Cellulaire Biochemie, Katholieke Universiteit te Leuven, Flanders, Belgium.
Eur J Biochem. 1990 Nov 13;193(3):675-80. doi: 10.1111/j.1432-1033.1990.tb19386.x.
In the yeast Saccharomyces cerevisiae, addition of glucose to cells grown under glucose-derepressed conditions induces a transient rise in the intracellular level of cAMP. This modulation requires functional elements of the cAMP-producing pathway, adenylate cyclase, ras proteins and the product of CDC25 gene. To determine whether or not the CDC25 gene product is a transducing element in the signal-transmission pathway leading from glucose to ras adenylate cyclase we have made use of the mutated allele RAS2Ile152 whose gene product uncouples the product of CDC25 from adenylate cyclase, but does not promotes other secondary phenotypes. The transient increase in cAMP is lost in cells lacking a functional CDC25 gene product, although they produce a normal amount of cAMP with the RAS2Ile152 gene. This result demonstrates the requirement of CDC25 for mediation of glucose signal transmission. The fact that cells grow normally on glucose in the absence of glucose-induced cAMP signaling confirms that this signaling pathway is not essential for growth on glucose. To further analyze the role of the CDC25 gene product we have made use of truncated versions of the gene. The results show that the C-terminal part of the gene alone is able to mediate glucose-induced activation of the RAS adenylate cyclase pathway.
在酿酒酵母中,向在葡萄糖去阻遏条件下生长的细胞添加葡萄糖会诱导细胞内cAMP水平短暂升高。这种调节需要cAMP产生途径的功能元件、腺苷酸环化酶、ras蛋白和CDC25基因的产物。为了确定CDC25基因产物是否是从葡萄糖到ras腺苷酸环化酶的信号传导途径中的转导元件,我们利用了突变等位基因RAS2Ile152,其基因产物使CDC25的产物与腺苷酸环化酶解偶联,但不促进其他次级表型。缺乏功能性CDC25基因产物的细胞中cAMP的短暂增加消失了,尽管它们用RAS2Ile152基因产生正常量的cAMP。这一结果证明了CDC25在介导葡萄糖信号传导中的必要性。在没有葡萄糖诱导的cAMP信号传导的情况下细胞在葡萄糖上正常生长这一事实证实了该信号传导途径对于在葡萄糖上生长不是必需的。为了进一步分析CDC25基因产物的作用,我们利用了该基因的截短版本。结果表明,仅该基因的C末端部分就能够介导葡萄糖诱导的RAS腺苷酸环化酶途径的激活。