title: Multiple effects of protein phosphatase 2A on nutrient-induced signalling in the yeast Saccharomyces cerevisiae.
abstract: The trehalose-degrading enzyme trehalase is activated upon addition of glucose to derepressed cells or in response to nitrogen source addition to nitrogen-starved glucose-repressed yeast (Saccharomyces cerevisiae) cells. Trehalase activation is mediated by phosphorylation. Inactivation involves dephosphorylation, as trehalase protein levels do not change upon multiple activation/inactivation cycles. Purified trehalase can be inactivated by incubation with protein phosphatase 2A (PP2A) in vitro. To test whether PP2A was involved in trehalase inactivation in vivo, we overexpressed the yeast PP2A isoform Pph22. Unexpectedly, the moderate (approximately threefold) overexpression of Pph22 that we obtained increased basal trehalase activity and rendered this activity unresponsive to the addition of glucose or a nitrogen source. Concomitant with higher basal trehalase activity, cells overexpressing Pph22 did not store trehalose efficiently and were heat sensitive. After the addition of glucose or of a nitrogen source to starved cells, Pph22-overexpressing cells showed a delayed exit from stationary phase, a delayed induction of ribosomal gene expression and constitutive repression of stress-regulated element-controlled genes. Deletion of the SCH9 gene encoding a protein kinase involved in nutrient-induced signal transduction restored glucose-induced trehalase activation in Pph22-overexpressing cells. Taken together, our results indicate that yeast PP2A overexpression leads to the activation of nutrient-induced signal transduction pathways in the absence of nutrients.
authors.affiliation: Cell and Molecular Signaling Laboratory, Intercollegiate Faculty of Biotechnology UG-MUG, Kladki 24, 80-822 Gdansk, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Sugajska
authors.forename: W
authors.initials: W
authors.lastname: Swiatek
authors.forename: P
authors.initials: P
authors.lastname: Zabrocki
authors.forename: I
authors.initials: I
authors.lastname: Geyskens
authors.forename: J M
authors.initials: JM
authors.lastname: Thevelein
authors.forename: S
authors.initials: S
authors.lastname: Zolnierowicz
authors.forename: S
authors.initials: S
authors.lastname: Wera
mesh_terms: D001483:Base Sequence; D020134:Catalytic Domain; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D008969:Molecular Sequence Data; D009584:Nitrogen / Q000378:metabolism; D019319:Okadaic Acid / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D054648:Protein Phosphatase 2; D012269:Ribosomal Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D015398:Signal Transduction; D014198:Trehalase / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D012269:Ribosomal Proteins; C031127:ribosomal protein L25; D019319:Okadaic Acid; D011494:Protein Kinases; C071569:SCH9 protein kinase; D010749:Phosphoprotein Phosphatases; D054648:Protein Phosphatase 2; D014198:Trehalase; D005947:Glucose; D009584:Nitrogen; D005990:Glycerol