Abstract
Genes involved in storage carbohydrate metabolism are coordinately induced when yeast cells are subjected to conditions of stress, or when they exit the exponential growth phase on glucose. We show that the STress Responsive Elements (STREs) present in the promoter of GSY2 are essential for gene activation under conditions of stress, but dispensable for gene induction and glycogen accumulation at the diauxic shift on glucose. Using serial promoter deletion, we found that the latter induction could not be attributed to a single cis -regulatory sequence, and present evidence that this mechanism depends on combinatorial transcriptional control by signalling pathways involving the protein kinases Pho85, Snf1 and PKA. Two contiguous regions upstream of the GSY2 coding region are necessary for negative control by the cyclin-dependent protein kinase Pho85, one of which is a 14-bp G/C-rich sequence. Positive control by Snf1 is mediated by Mig1p, which acts indirectly on the distal part of the GSY2 promoter. The PKA pathway has the most pronounced effect on GSY2, since transcription of this gene is almost completely abolished in an ira1ira2 mutant strain in which PKA is hyperactive. The potent negative effect of PKA is dependent upon a branched pathway involving the transcription factors Msn2/Msn4p and Sok2p. The SOK2 branch was found to be effective only under conditions of high PKA activity, as in a ira1ira2 mutant, and this effect was independent of Msn2/4p. The Msn2/4p branch, on the other hand, positively controls GSY2 expression directly through the STREs, and indirectly via a factor that still remains to be discovered. In summary, this study shows that the transcription of GSY2 is regulated by several different signalling pathways which reflect the numerous factors that influence glycogen synthesis in yeast, and suggests that the PKA pathway must be deactivated to allow gene induction at the diauxic shift.
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Acknowledgements
We thank M. Jacquet (University of Paris-Sud, France), J. Cannon (University of Columbia, Missouri, USA), F. Estruch (CSIC, Valencia, Spain), H. Ronne (Uppsala, Sweden), Martine Collard (University of Geneva, Switzerland) and A. Sakai (University of Tokyo, Japan) for kind provision of yeast strains and/or plasmids. We are grateful to Kelly Tatchell and Lucy Robinson (University of Shreveport, Louisiana, USA) and to our colleagues for critical reading of the manuscript. This work was supported in part by the ‘Microbiology and Pathogenicity’ program of the French Ministry of Education. BE was supported by a doctoral grant from the French Ministry of Education and Research.
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Enjalbert, B., Parrou, J.L., Teste, M.A. et al. Combinatorial control by the protein kinases PKA, PHO85 and SNF1 of transcriptional induction of the Saccharomyces cerevisiae GSY2 gene at the diauxic shift. Mol Genet Genomics 271, 697–708 (2004). https://doi.org/10.1007/s00438-004-1014-8
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DOI: https://doi.org/10.1007/s00438-004-1014-8