title: Involvement of distinct G-proteins, Gpa2 and Ras, in glucose- and intracellular acidification-induced cAMP signalling in the yeast Saccharomyces cerevisiae.
abstract: 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 ira1Delta ira2Delta 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 Galpha-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) with adenylate cyclase apparently prevents its activation by both agonists.
authors.affiliation: Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven, Flanders, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Colombo
authors.forename: P
authors.initials: P
authors.lastname: Ma
authors.forename: L
authors.initials: L
authors.lastname: Cauwenberg
authors.forename: J
authors.initials: J
authors.lastname: Winderickx
authors.forename: M
authors.initials: M
authors.lastname: Crauwels
authors.forename: A
authors.initials: A
authors.lastname: Teunissen
authors.forename: D
authors.initials: D
authors.lastname: Nauwelaers
authors.forename: J H
authors.initials: JH
authors.lastname: de Winde
authors.forename: M F
authors.initials: MF
authors.lastname: Gorwa
authors.forename: D
authors.initials: D
authors.lastname: Colavizza
authors.forename: J M
authors.initials: JM
authors.lastname: Thevelein
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D000242:Cyclic AMP / Q000096:biosynthesis / Q000378:metabolism*; D015536:Down-Regulation; D005656:Fungal Proteins / Q000378:metabolism*; D044385:GTP-Binding Protein alpha Subunits; D019204:GTP-Binding Proteins / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D020962:Heterotrimeric GTP-Binding Proteins; D006863:Hydrogen-Ion Concentration; D010749:Phosphoprotein Phosphatases / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D020668:rap GTP-Binding Proteins; D018631:ras Proteins / Q000378:metabolism*; D020666:ras-GRF1
chemical_list: D018797:Cell Cycle Proteins; D005656:Fungal Proteins; D044385:GTP-Binding Protein alpha Subunits; D029701:Saccharomyces cerevisiae Proteins; D020666:ras-GRF1; D000242:Cyclic AMP; D010749:Phosphoprotein Phosphatases; D019204:GTP-Binding Proteins; C083907:Gpa2 protein, S cerevisiae; D020962:Heterotrimeric GTP-Binding Proteins; D020668:rap GTP-Binding Proteins; D018631:ras Proteins; D000262:Adenylyl Cyclases; D005947:Glucose