The activity of adenylate cyclase in the yeast Saccharomyces cerevisiae is controlled by two G-protein systems, the Ras proteins and the Gα protein Gpa2. Glucose activation of cAMP synthesis is thought to be mediated by Gpa2 and its G-protein-coupled receptor Gpr1. Using a sensitive GTP-loading assay for Ras2 we demonstrate that glucose addition also triggers a fast increase in the GTP loading state of Ras2 concomitant with the glucose-induced increase in cAMP. This increase is severely delayed in a strain lacking Cdc25, the guanine nucleotide exchange factor for Ras proteins. Deletion of the Ras-GAPs IRA2 (alone or with IRA1) or the presence of RAS2<sup>Val19</sup> allele causes constitutively high Ras GTP loading that no longer increases upon glucose addition. The glucose-induced increase in Ras2 GTP-loading is not dependent on Gpr1 or Gpa2. Deletion of these proteins causes higher GTP loading indicating that the two G-protein systems might directly or indirectly interact. Because deletion of GPR1 or GPA2 reduces the glucose-induced cAMP increase the observed enhancement of Ras2 GTP loading is not sufficient for full stimulation of cAMP synthesis. Glucose phosphorylation by glucokinase or the hexokinases is required for glucose-induced Ras2 GTP loading. These results indicate that glucose phosphorylation might sustain activation of cAWDP synthesis by enhancing Ras2 GTP loading likely through inhibition of the Ira proteins. Strains with reduced feedback inhibition on cAMP synthesis also display elevated basal and induced Ras2 GTP loading consistent with the Ras2 protein acting as a target of the feedback-inhibition mechanism.
Colombo, S., Ronchetti, D., Thevelein, J.M., Winderickx, J., & Martegani, E. (2004). Activation state of the Ras2 protein and glucose-induced signalling in Saccharomyces cerevisiae. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 279, 46715-46722 [10.1074/jbc.M405136200].
|Citazione:||Colombo, S., Ronchetti, D., Thevelein, J.M., Winderickx, J., & Martegani, E. (2004). Activation state of the Ras2 protein and glucose-induced signalling in Saccharomyces cerevisiae. THE JOURNAL OF BIOLOGICAL CHEMISTRY, 279, 46715-46722 [10.1074/jbc.M405136200].|
|Tipo:||Articolo in rivista - Articolo scientifico|
|Carattere della pubblicazione:||Scientifica|
|Presenza di un coautore afferente ad Istituzioni straniere:||Si|
|Titolo:||Activation state of the Ras2 protein and glucose-induced signalling in Saccharomyces cerevisiae|
|Autori:||Colombo, S; Ronchetti, D; Thevelein, JM; Winderickx, J; Martegani, E|
|Data di pubblicazione:||2004|
|Rivista:||THE JOURNAL OF BIOLOGICAL CHEMISTRY|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1074/jbc.M405136200|
|Appare nelle tipologie:||01 - Articolo su rivista|