Abstract
The phosphate regulatory mechanism in yeast, known as the PHO pathway, is regulated by inorganic phosphate to control the expression of genes involved in the acquisition of phosphate from the medium. This pathway is also reported to contribute to other nutritional responses and as such it affects several phenotypic characteristics known also to be regulated by protein kinase A, including the transcription of genes involved in the general stress response and trehalose metabolism. We now demonstrate that transcription of post-diauxic shift (PDS)-controlled stress-responsive genes is solely regulated by the Pho85–Pho80 complex, whereas regulation of trehalose metabolism apparently involves several Pho85 cyclins. Interestingly, both read-outs depend on Pho81 but, while the previously described minimum domain of Pho81 is sufficient to sustain phosphate-regulated transcription of PHO genes, full-length Pho81 is required to control trehalose metabolism and the PDS targets. Consistently, neither the expression control of stress-regulated genes nor the trehalose metabolism relies directly on Pho4. Finally, we present data supporting that the PHO pathway functions in parallel to the fermentable growth medium- or Sch9-controlled pathway and that both pathways may share the protein kinase Rim15, which was previously reported to play a central role in the integration of glucose, nitrogen and amino acid availability.
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Aerne BL, Johnson AL, Toyn JH, Johnston LH (1998) Swi5 controls a novel wave of cyclin synthesis in late mitosis. Mol Biol Cell 9:945–956
Brachmann CB, Davies A, Cost GJ, Caputo E, Li J, Hieter P, Boeke JD (1998) Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14:115–132
Cameroni E, Hulo N, Roosen J, Winderickx J, De Virgilio C (2004) The novel yeast PAS kinase Rim15 orchestrates G(0)-associated antioxidant defense mechanisms. Cell Cycle 3:462–468
Carroll AS, O’Shea EK (2002) Pho85 and signaling environmental conditions. Trends Biochem Sci 27:87–93
Colombo S, Ma P, Cauwenberg L, Winderickx J, Crauwels M, Teunissen A, Nauwelaers D, Winde JH de, Gorwa MF, Colavizza D, Thevelein JM (1998) Involvement of distinct G-proteins, Gpa2 and Ras, in glucose- and intracellular acidification-induced cAMP signalling in the yeast Saccharomyces cerevisiae. EMBO J 17:3326–3341
Crauwels M, Donaton MC, Pernambuco MB, Winderickx J, Winde JH de, Thevelein JM (1997) The Sch9 protein kinase in the yeast Saccharomyces cerevisiae controls cAPK activity and is required for nitrogen activation of the fermentable-growth-medium-induced (FGM) pathway. Microbiology 143:2627–2637
Creasy CL, Madden SL, Bergman LW (1993) Molecular analysis of the PHO81 gene of Saccharomyces cerevisiae. Nucleic Acids Res 21:1975–1982
Donaton MC, Holsbeeks I, Lagatie O, Van Zeebroeck G, Crauwels M, Winderickx J, Thevelein JM (2003) The Gap1 general amino acid permease acts as an amino acid sensor for activation of protein kinase A targets in the yeast Saccharomyces cerevisiae. Mol Microbiol 50:911–929
Durnez P, Pernambuco MB, Oris E, Arguelles JC, Mergelsberg H, Thevelein JM (1994) Activation of trehalase during growth induction by nitrogen sources in the yeast Saccharomyces cerevisiae depends on the free catalytic subunits of cAMP-dependent protein kinase, but not on functional Ras proteins. Yeast 10:1049–1064
Espinoza FH, Ogas J, Herskowitz I, Morgan DO (1994) Cell cycle control by a complex of the cyclin HCS26 (PCL1) and the kinase PHO85. Science 266:1388–1391
Flick JS, Thorner J (1993) Genetic and biochemical characterization of a phosphatidylinositol-specific phospholipase C in Saccharomyces cerevisiae. Mol Cell Biol 13:5861–5876
Flick JS, Thorner J (1998) An essential function of a phosphoinositide-specific phospholipase C is relieved by inhibition of a cyclin-dependent protein kinase in the yeast Saccharomyces cerevisiae. Genetics 148:33–47
Francois J, Parrou JL (2001) Reserve carbohydrates metabolism in the yeast Saccharomyces cerevisiae. FEMS Microbiol Rev 25:125–145
Gilliquet V, Berben G (1993) Positive and negative regulators of the Saccharomyces cerevisiae ‘PHO system’ participate in several cell functions. FEMS Microbiol Lett 108:333–339
Giots F, Donaton MC, Thevelein JM (2003) Inorganic phosphate is sensed by specific phosphate carriers and acts in concert with glucose as a nutrient signal for activation of the protein kinase A pathway in the yeast Saccharomyces cerevisiae. Mol Microbiol 47:1163–1181
Hirimburegama K, Durnez P, Keleman J, Oris E, Vergauwen R, Mergelsberg H, Thevelein JM (1992) Nutrient-induced activation of trehalase in nutrient-starved cells of the yeast Saccharomyces cerevisiae: cAMP is not involved as second messenger. J Gen Microbiol 138:2035–2043
Ho Y, Gruhler A, Heilbut A, Bader GD, Moore L, Adams SL, Millar A, Taylor P, Bennett K, Boutilier K, Yang L, Wolting C, Donaldson I, Schandorff S, Shewnarane J, Vo M, Taggart J, Goudreault M, Muskat B, Alfarano C, Dewar D, Lin Z, Michalickova K, Willems AR, Sassi H, Nielsen PA, Rasmussen KJ, Andersen JR, Johansen LE, Hansen LH, Jespersen H, Podtelejnikov A, Nielsen E, Crawford J, Poulsen V, Sorensen BD, Matthiesen J, Hendrickson RC, Gleeson F, Pawson T, Moran MF, Durocher D, Mann M, Hogue CW, Figeys D, Tyers M (2002) Systematic identification of protein complexes in Saccharomyces cerevisiae by mass spectrometry. Nature 415:180–183
Huang D, Farkas I, Roach PJ (1996) Pho85p, a cyclin-dependent protein kinase, and the Snf1p protein kinase act antagonistically to control glycogen accumulation in Saccharomyces cerevisiae. Mol Cell Biol 16:4357–4365
Huang D, Moffat J, Wilson WA, Moore L, Cheng C, Roach PJ, Andrews B (1998) Cyclin partners determine Pho85 protein kinase substrate specificity in vitro and in vivo: control of glycogen biosynthesis by Pcl8 and Pcl10. Mol Cell Biol 18:3289–3299
Huang S, Jeffery DA, Anthony MD, O’Shea EK (2001) Functional analysis of the cyclin-dependent kinase inhibitor Pho81 identifies a novel inhibitory domain. Mol Cell Biol 21:6695–6705
Huang D, Moffat J, Andrews B (2002) Dissection of a complex phenotype by functional genomics reveals roles for the yeast cyclin-dependent protein kinase Pho85 in stress adaptation and cell integrity. Mol Cell Biol 22:5076–5088
Kaffman A, Herskowitz I, Tjian R, O’Shea EK (1994) Phosphorylation of the transcription factor PHO4 by a cyclin-CDK complex, PHO80-PHO85. Science 263:1153–1156
Kaffman A, Rank NM, O’Neill EM, Huang LS, O’Shea EK (1998a) The receptor Msn5 exports the phosphorylated transcription factor Pho4 out of the nucleus. Nature 396:482–486
Kaffman A, Rank NM, O’Shea EK (1998b) Phosphorylation regulates association of the transcription factor Pho4 with its import receptor Pse1/Kap121. Genes Dev 12:2673–2683
Knight JP, Daly TM, Bergman LW (2004) Regulation by phosphorylation of Pho81p, a cyclin-dependent kinase inhibitor in Saccharomyces cerevisiae. Curr Genet 46:10–19
Komeili A, O’Shea EK (1999) Roles of phosphorylation sites in regulating activity of the transcription factor Pho4. Science 284:977–980
Lee M, O’Regan S, Moreau JL, Johnson AL, Johnston LH, Goding CR (2000) Regulation of the Pcl7-Pho85 cyclin-cdk complex by Pho81. Mol Microbiol 38:411–422
Lenburg ME, O’Shea EK (1996) Signaling phosphate starvation. Trends Biochem Sci 21:383–387
Lillie SH, Pringle JR (1980) Reserve carbohydrate metabolism in Saccharomyces cerevisiae: responses to nutrient limitation. J Bacteriol 143:1384–1394
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Measday V, Moore L, Ogas J, Tyers M, Andrews B (1994) The PCL2 (ORFD)-PHO85 cyclin-dependent kinase complex: a cell cycle regulator in yeast. Science 266:1391–1395
Measday V, Moore L, Retnakaran R, Lee J, Donoviel M, Neiman AM, Andrews B (1997) A family of cyclin-like proteins that interact with the Pho85 cyclin-dependent kinase. Mol Cell Biol 17:1212–1223
Moffat J, Huang D, Andrews B (2000) Functions of Pho85 cyclin-dependent kinases in budding yeast. Prog Cell Cycle Res 4:97–106
Neves MJ, Terenzi HF, Leone FA, Jorge JA (1994) Quantification of trehalose in biological samples with a conidial trehalase from the thermophilic fungus Humicola grisea var Thermoidea. World J Microbiol Biotechnol 10:17–19
Nishizawa M, Katou Y, Shirahige K, Toh EA (2004) Yeast Pho85 kinase is required for proper gene expression during the diauxic shift. Yeast 21:903–918
Ogas J, Andrews BJ, Herskowitz I (1991) Transcriptional activation of CLN1, CLN2, and a putative new G1 cyclin (HCS26) by SWI4, a positive regulator of G1-specific transcription. Cell 66:1015–1026
Ogawa N, Noguchi K, Yamashita Y, Yasuhara T, Hayashi N, Yoshida K, Oshima Y (1993) Promoter analysis of the PHO81 gene encoding a 134 kDa protein bearing ankyrin repeats in the phosphatase regulon of Saccharomyces cerevisiae. Mol Gen Genet 238:444–454
Ogawa N, Noguchi K, Sawai H, Yamashita Y, Yompakdee C, Oshima Y (1995) Functional domains of Pho81p, an inhibitor of Pho85p protein kinase, in the transduction pathway of Pi signals in Saccharomyces cerevisiae. Mol Cell Biol 15:997–1004
Ogawa N, DeRisi J, Brown PO (2000) New components of a system for phosphate accumulation and polyphosphate metabolism in Saccharomyces cerevisiae revealed by genomic expression analysis. Mol Biol Cell 11:4309–4321
O’Neill EM, Kaffman A, Jolly ER, O’Shea EK (1996) Regulation of PHO4 nuclear localization by the PHO80–PHO85 cyclin-CDK complex. Science 271:209–212
Pedruzzi I, Burckert N, Egger P, De Virgilio C (2000) Saccharomyces cerevisiae Ras/cAMP pathway controls post-diauxic shift element-dependent transcription through the zinc finger protein Gis1. EMBO J 19:2569–2579
Pedruzzi I, Dubouloz F, Cameroni E, Wanke V, Roosen J, Winderickx J, De Virgilio C (2003) TOR and PKA signaling pathways converge on the protein kinase Rim15 to control entry into G0. Mol Cell 12:1607–1613
Persson BL, Lagerstedt JO, Pratt JR, Pattison-Granberg J, Lundh K, Shokrollahzadeh S, Lundh F (2003) Regulation of phosphate acquisition in Saccharomyces cerevisiae. Curr Genet 43:225–244
Reinders A, Burckert N, Boller T, Wiemken A, De Virgilio C (1998) Saccharomyces cerevisiae cAMP-dependent protein kinase controls entry into stationary phase through the Rim15p protein kinase. Genes Dev 12:2943–2955
Roosen J, Oesterhelt C, Pardons K, Swinnen E, Winderickx J (2004) Integration of nutrient signalling pathways in the yeast Saccharomyces cerevisiae. In: Winderickx J, Taylor PM (eds) Nutrient-induced responses in eukaryotic cells. Topics Curr Genet 2004:277–318
Roosen J, Engelen K, Marchal K, Mathys J, Griffioen G, Cameroni E, Thevelein JM, De Virgilio C, De Moor B, Winderickx J (2005) PKA and Sch9 control a molecular switch important for the proper adaptation to nutrient availability. Mol Microbiol 55:862–880
Schneider KR, Smith RL, O’Shea EK (1994) Phosphate-regulated inactivation of the kinase PHO80-PHO85 by the CDK inhibitor PHO81. Science 266:122–126
Shemer R, Meimoun A, Holtzman T, Kornitzer D (2002) Regulation of the transcription factor Gcn4 by Pho85 cyclin PCL5. Mol Cell Biol 22:5395–5404
Tennyson CN, Lee J, Andrews BJ (1998) A role for the Pcl9-Pho85 cyclin-cdk complex at the M/G1 boundary in Saccharomyces cerevisiae. Mol Microbiol 28:69–79
Thevelein JM (1994) Signal transduction in yeast. Yeast 10:1753–1790
Thevelein JM, Winde JH de (1999) Novel sensing mechanisms and targets for the cAMP-protein kinase A pathway in the yeast Saccharomyces cerevisiae. Mol Microbiol 33:904–918
Timblin BK, Tatchell K, Bergman LW (1996) Deletion of the gene encoding the cyclin-dependent protein kinase Pho85 alters glycogen metabolism in Saccharomyces cerevisiae. Genetics 143:57–66
Waters NC, Knight JP, Creasy CL, Bergman LW (2004) The yeast Pho80-Pho85 cyclin-CDK complex has multiple substrates. Curr Genet 46:1–9
Wilson WA, Mahrenholz AM, Roach PJ (1999) Substrate targeting of the yeast cyclin-dependent kinase Pho85p by the cyclin Pcl10p. Mol Cell Biol 19:7020–7030
Wilson WA, Wang Z, Roach PJ (2005) Regulation of yeast glycogen phosphorylase by the cyclin-dependent protein kinase Pho85p. Biochem Biophys Res Commun 329:161–167
Winderickx J, Winde JH de, Crauwels M, Hino A, Hohmann S, Van Dijck P, Thevelein JM (1996) Regulation of genes encoding subunits of the trehalose synthase complex in Saccharomyces cerevisiae: novel variations of STRE-mediated transcription control? Mol Gen Genet 252:470–482
Winderickx J, Holsbeeks I, Lagatie O, Giots F, Thevelein JM, De Winde H (2003) From feast to famine; adaptation to nutrient availability in yeast. In: Hohmann S, Mager PWH (eds) Yeast stress responses. Topics Curr Genet 2003:305–386
Zahringer H, Holzer H, Nwaka S (1998) Stability of neutral trehalase during heat stress in Saccharomyces cerevisiae is dependent on the activity of the catalytic subunits of cAMP-dependent protein kinase, Tpk1 and Tpk2. Eur J Biochem 255:544–551
Zahringer H, Thevelein JM, Nwaka S (2000) Induction of neutral trehalase Nth1 by heat and osmotic stress is controlled by STRE elements and Msn2/Msn4 transcription factors: variations of PKA effect during stress and growth. Mol Microbiol 35:397–406
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This work was supported by a fellowship from the Fund of Scientific Research in Flanders (FWO-Vlaanderen) to E.S. and by grants from FWO-Vlaanderen and the research fund of K.U. Leuven
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Swinnen, E., Rosseels, J. & Winderickx, J. The minimum domain of Pho81 is not sufficient to control the Pho85–Rim15 effector branch involved in phosphate starvation-induced stress responses. Curr Genet 48, 18–33 (2005). https://doi.org/10.1007/s00294-005-0583-3
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DOI: https://doi.org/10.1007/s00294-005-0583-3