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PMID: 9628870 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

The EMBO journal ·Vol. 17 ·No. 12 ·1998-06-15 ·Pages 3326-41

Colombo S, Ma P, Cauwenberg L, Winderickx J, Crauwels M, Teunissen A, Nauwelaers D, de Winde JH, Gorwa MF, Colavizza D, Thevelein JM

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.

MeSH Terms
Adenylyl Cyclases/metabolism Cell Cycle Proteins/metabolism Cyclic AMP/biosynthesis,metabolism Down-Regulation Fungal Proteins/metabolism GTP-Binding Protein alpha Subunits GTP-Binding Proteins/metabolism Glucose/metabolism Heterotrimeric GTP-Binding Proteins Hydrogen-Ion Concentration Phosphoprotein Phosphatases/metabolism Saccharomyces cerevisiae/enzymology,metabolism Saccharomyces cerevisiae Proteins Signal Transduction rap GTP-Binding Proteins ras Proteins/metabolism ras-GRF1
Chemicals
Cell Cycle Proteins Fungal Proteins GTP-Binding Protein alpha Subunits Saccharomyces cerevisiae Proteins ras-GRF1 Cyclic AMP Phosphoprotein Phosphatases GTP-Binding Proteins Gpa2 protein, S cerevisiae Heterotrimeric GTP-Binding Proteins rap GTP-Binding Proteins ras Proteins Adenylyl Cyclases Glucose
Authors & Affiliations
11 authors, click to expand affiliations / ORCID
Colombo S
Laboratorium voor Moleculaire Celbiologie, Katholieke Universiteit Leuven, Flanders, Belgium.
Ma P
Cauwenberg L
Winderickx J
Crauwels M
Teunissen A
Nauwelaers D
de Winde J H
Gorwa M F
Colavizza D
Thevelein J M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.12.3326
Published
1998-06-15
Pages
3326-41
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170671
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.12.3326 PMC Full Text View journal details
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