Involvement of distinct G-proteins, Gpa2 and Ras, in glucose- and intracellular acidification-induced cAMP signalling in the yeast Saccharomyces cerevisiae

被引:247
作者
Colombo, S
Ma, PS
Cauwenberg, L
Winderickx, J
Crauwels, M
Teunissen, A
Nauwelaers, D
de Winde, JH
Gorwa, MF
Colavizza, D
Thevelein, JM
机构
[1] Katholieke Univ Leuven, Lab Mol Celbiol, B-3001 Leuven, Flanders, Belgium
[2] Tech Univ Darmstadt, Inst Mikrobiol & Genet, D-64287 Darmstadt, Germany
[3] Lesaffre Dev SA, F-57906 Marcq En Baroeul, France
关键词
acidification; cAMP signalling; Gpa2; glucose; Ras;
D O I
10.1093/emboj/17.12.3326
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Adenylate cyclase activity in Saccharomyces cerevisiae is dependent on Ras proteins. Both addition of glucose to glucose-deprived (derepressed) cells and intracellular acidification trigger an increase in the cAMP level in vivo, We show that intracellular acidification, but not glucose, causes an increase in the GTP/GDP ratio on the Ras proteins independent of Cdc25 and Sdc25. Deletion of the GTPase-activating proteins Ira1 and Ira2, or expression of the RAS2(val19) allele, causes an enhanced GTP/GDP basal ratio and abolishes the intracellular acidification-induced increase. In the iral Delta ira2 Delta strain, intracellular acidification still triggers a cAMP increase. Glucose also did not cause an increase in.the GTP/GDP ratio in a strain with reduced feedback inhibition of cAMP synthesis. Further investigation indicated that feedback inhibition by cAPK on cAMP synthesis acts independently of changes in the GTP/GDP ratio on Ras. Stimulation by glucose was dependent on the G alpha-protein Gpa2, whose deletion confers the typical phenotype associated with a reduced cAMP level: higher heat resistance, a higher level of trehalose and glycogen and elevated expression of STRE-controlled genes. However, the typical fluctuation in these characteristics during diauxic growth on glucose was still present. Overexpression of Ras2(val19) inhibited both the acidification- and glucose-induced cAMP increase even in a protein kinase A-attenuated strain. Our results suggest that intracellular acidification stimulates cAMP synthesis in vivo at least through activation of the Ras proteins, while glucose acts through the Gpa2 protein. Interaction of Ras2(val19),vith adenylate cyclase apparently prevents its activation by both agonists.
引用
收藏
页码:3326 / 3341
页数:16
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