Molecular and General Genetics MGG

, Volume 252, Issue 4, pp 470–482

Regulation of genes encoding subunits of the trehalose synthase complex inSaccharomyces cerevisiae: novel variations of STRE-mediated transcription control?

  • J. Winderickx
  • J. H. de Winde
  • M. Crauwels
  • A. Hino
  • S. Hohmann
  • P. Van Dijck
  • J. M. Thevelein
Original Paper


Saccharomyces cerevisiae cells show under suboptimal growth conditions a complex response that leads to the acquisition of tolerance to different types of environmental stress. This response is characterised by enhanced expression of a number of genes which contain so-called stress-responsive elements (STREs) in their promoters. In addition, the cells accumulate under suboptimal conditions the putative stress protectant trehalose. In this work, we have examined the expression of four genes encoding subunits of the trehalose synthase complex,GGS1/TPS1, TPS2, TPS3 andTSL1. We show that expression of these genes is coregulated under stress conditions. Like for many other genes containing STREs, expression of the trehalose synthase genes is also induced by heat and osmotic stress and by nutrient starvation, and negatively regulated by the Ras-cAMP pathway. However, during fermentative growth onlyTSL1 shows an expression pattern like that of the STRE-controlled genesCTT1 andSSA3, while expression of the three other trehalose synthase genes is only transiently down-regulated. This difference in expression might be related to the known requirement of trehalose biosynthesis for the control of yeast glycolysis and hence for fermentative growth. We conclude that the mere presence in the promoter of (an) active STRE(s) does not necessarily imply complete coregulation of expression. Additional mechanisms appear to fine tune the activity of STREs in order to adapt the expression of the downstream genes to specific requirements.

Key words

Trehalose synthase complex Stress-responsive element (STRE) Stress response Nutrients Ras-cAMP pathway 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. App H, Holzer H (1989) Purification and characterisation of neutral trehalase from the yeastABYS1 mutant. J Biol Chem 264:17583–17588Google Scholar
  2. Argülles JC (1994) Heat-shock response in a yeasttps1 mutant deficient in trehalose synthesis. FEBS Lett 350:266–270Google Scholar
  3. Arguelles JC, Myombi K, Van Aelst L, Vanhalewyn M, Jans AWH, Thevelein JM (1990) Absence of glucose-induced cAMP signalling in theSaccharomyces cerevisiae mutantscat1 andcat3, which are deficient in derepression of glucose-repressible proteins. Arch Microbiol 154:199–205Google Scholar
  4. Attfiel PV, Ramn A, Norhcott CJ (1992) Construction ofSaccharomyces cerevisiae strains that accumulate relatively low levels of trehalose, and their application in testing the contribution of the disaccharide to stress tolerance. FEMS Microbiol Lett 94:271–276Google Scholar
  5. Bell W, Klaassen P, Ohnacker M, Boller T, Herweijer M, Schoppink P, van der Zee P, Wiemken A (1992) Characterisation of the 56-kDa subunit of trehalose-6-phosphate synthase and cloning of its gene reveal its identity with the product ofClF1, a regulator of carbon catabolite inactivation. Eur J Biochem 209:951–959Google Scholar
  6. Beullens M, Mbonyi K, Geerts L, Gladines D, Detremerie K, Jans AWH, Thevelein JM (1988) Studies on the mechanism of the glucose-induced cAMP-signal in glycolysis- and glucose-repression mutants of the yeastSaccharomyces cerevisiae. Eur J Biochem 172:227–231Google Scholar
  7. Boorstein WR, Craig EA (1990) Regulation of yeastHSP70 gene by a cAMP responsive transcription control element. EMBO J 9:2543–2553Google Scholar
  8. Bossier P, Fernandez D, Rocha D, Rodigues-Pousada C (1993) Overexpression ofYAP2, coding for a new yAP protein, andYAP1 inSaccharomyces cerevisiae alleviates growth inhibition caused by 1,10-phenanthroline. J Biol Chem 268:23640–23645Google Scholar
  9. Cannon JF, Pringle JR, Fiechter A, Khalil M (1994) Characterisation of glycogen-deficientglc mutants ofSaccharomyces cerevisiae. Genetics 136:485–503Google Scholar
  10. Coote PJ, Jones MV, Edgar K, Cole MB (1992)TPK gene products mediate cAMP-independent thermotolerance inSaccharomyces cerevisiae. J Gen Microbiol 138:2551–2557Google Scholar
  11. Coutinho CC, Silva JT, Panek AD (1992) Trehalase activity and its regulation during growth ofSaccharomyces cerevisiae. Biochem Int 26:521–530Google Scholar
  12. De Virgilio C, Bürckert N, Boller T, Wiemken A (1991) A method to study the rapid phosphorylation-related modulation of neutral trehalase activity by temperature shifts in yeast. FEBS Lett 291:355–358Google Scholar
  13. De Virgilio C, Bürckert N, Bell W, Jenö P, Boller T, Wiemken A (1993) Disruption ofTPS2, the gene encoding the 100-kDa subunit of the trehalose-6-phosphate synthase/phosphatase complex inSaccharomyces cerevisiae, causes accumulation of trehalose-6-phosphate and loss of trehalose-6-phosphate phosphatase activity. Eur J Biochem 212:315–323Google Scholar
  14. Domdey H, Apostol B, Lin R-J, Newman A, Brody E, Abelson J (1984) Lariat structures are in vivo intermediates in yeast pre-mRNA splicing. Cell 39:611–621Google Scholar
  15. Durnez P, Oris E, Argüelles JC, Mergelsberg H, Thevelein JM (1994) Activation of trehalase during growth induction by nitrogen sources in the yeastSaccharomyces cerevisiae depends on the free catalytic subunits of cAMP-dependent protein kinase. Yeast 10:1049–1064Google Scholar
  16. Entian K-D (1981) A carbon catabolite repression mutant ofSaccharomyces cerevisiae with elevated hexokinase activity: evidence for regulatory control of hexokinase PH synthesis. Mol Gen Genet 184:278–282Google Scholar
  17. François J, Eraso P, Gancedo C (1987) Changes in the concentration of cAMP, fructose-2,6-biphosphate and related metabolites and enzymes inSaccharomyces cerevisiae during growth on glucose. Eur J Biochem 164:369–373Google Scholar
  18. François J, Neves M-J, Hers H-G (1991) The control of trehalose biosynthesis inSaccharomyces cerevisiae: evidence for a catabolite inactivation and repression of trehalose-6-phosphate synthase and trehalose-6-phosphate phosphatase. Yeast 7:575–587Google Scholar
  19. Gallwitz D, Sures I (1980) Structure of a split yeast gene: complete nucleotide sequence of the actin gene inSaccharomyces cerevisiae. Proc Natl Acad Sci USA 77:2546–2550Google Scholar
  20. Gonzalez MI, Stucka R, Blazquez MA, Feldmann H, Gancedo C (1992) Molecular cloning ofClF1, a yeast gene necessary for growth on glucose. Yeast 8:183–192Google Scholar
  21. Gounalaki N, Thireos G (1994) Yap1p, a yeast transcription activator that mediates multidrug resistance, regulates the metabolic stress response. EMBO J 13:4036–4041Google Scholar
  22. 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 yeastSaccharomyces cerevisiae: cAMP is not involved as second messenger. J Gen Microbiol 138:2035–2043Google Scholar
  23. Hohmann S, Neves MJ, de Koning W, Alijo R, Ramos J, Thevelein JM (1993) The growth and signalling defects in theggs1 (fdp1/byp1) deletion mutant on glucose are suppressed by a deletion of the gene encoding hexokinase PH. Cum Genet 23:281–289Google Scholar
  24. Hohmann S, Van Dijck P, Luyten K, Thevelein JM (1994) Thebyp1-3 allele of theSaccharomyces cerevisiae GGS1/TPS1 gene and its multicopy suppressor tRNAGLN (CAG): Ggs1/Tps1 levels restraining growth on fermentable sugars and trehalose accumulation. Curr Genet 26:295–301Google Scholar
  25. Hottiger T, Boller T, Wiemken A (1987a) Rapid changes of heat and dissication tolerance correlated with changes of trehalose content inSaccharomyces cerevisiae cells subjected to temperature shifts. FEBS Lett 220:113–115Google Scholar
  26. Hottiger T, Schmutz P, Wiemken A (1987b) Heat-induced accumulation and futile cycling of trehalose inSaccharomyces cerevisiae. J Bacteriol 169:5518–5522Google Scholar
  27. Hottiger T, Boller T, Wiemken A (1989) Correlation of trehalose content and heat resistance in yeast altered in the Ras/adenylate cyclase pathway: is trehalose a thermoprotectant? FEBS Lett 255:431–434Google Scholar
  28. Jakobsen BK, Pelham HRB (1988) Constitutive binding of yeast heat shock factor to DNA in vivo. Mol Cell Biol 8:5040–5042Google Scholar
  29. Kobayashi N, McEntee K (1990) Evidence for a heat shock transcription factor-independent mechanism for heat shock induction of transcription inSaccharomyces cerevisiae. Proc Nat Acad Sci USA 87:6550–6554Google Scholar
  30. Kobayashi N, McEntee K (1993) Identification of cis and trans components of a novel heat shock stress regulatory pathway inSaccharomyces cerevisiae Mol Cell Biol 13:248–256Google Scholar
  31. Lillie SH, Pringle JR (1980) Reverse carbohydrate metabolism inSaccharomyces cerevisiae. Response to nutrient limitation. J Bacteriol 143:1384–1394Google Scholar
  32. Mager WH, Moradas Ferreira P (1983) Stress response in yeast. Biochem J 290:1–13Google Scholar
  33. Mager WH, De Kruijff AJJ (1995) Stress-induced transcriptional activation. Microbiol Rev 59:506–531Google Scholar
  34. Marchler G, Schüller C, Adam G, Ruis H (1993)Saccharomyces cerevisiae UAS element controlled by protein kinase A activates transcription in response to a variety of stress conditions. EMBO J 12:1997–2003Google Scholar
  35. Matsumoto K, Uno I, Ishikawa T (1985) Genetic analysis of the role of cAMP in yeast. Yeast 1:15–24Google Scholar
  36. McDougall J, Kaasen I, Ström AR (1993) A yeast gene for trehalose-6-phosphate synthase and its complementation of anEscherichia coli otsA mutant. FEMS Microbiol Lett 107:25–30Google Scholar
  37. Myers AM, Tzagoloff A, Kinney DM, Lusty CJ (1986) Yeast shuttle and integrative vectors with multiple cloning sites suitable for construction oflacZ fusions. Gene 45:299–310Google Scholar
  38. Neves M-J, François J (1992) On the mechanism by which a heat shock induces trehalose accumulation inSaccharomyces cerevisiae. Biochem J 288:859–864Google Scholar
  39. Neves M-J, Hohmann S, Bell W, Dumortier F, Luyten K, Ramos J, Cobbaert P, de Koning W, Kaneva Z, Thevelein JM (1995) Control of glucose influx into glycolysis and pleiotropic effects studied in different isogenic sets ofSaccharomyces cerevisiae mutants in trehalose biosynthesis. Curr Genet 27:306–308Google Scholar
  40. Nikawa J, Cameron S, Toda T, Ferguson KW, Wigler M (1987) Rigorous feedback control of cAMP levels inSaccharomyces cerevisiae. Genes Dev 1:931–937Google Scholar
  41. Nwaka S, Kopp M, Burgert M, Deuchler I, Kienle I, Holzer H (1984) Is thermotolerance of yeast dependent on trehalose accumulation? FEBS Lett 344:255–228Google Scholar
  42. Nwaka S, Kopp M, Holzer H (1995a) Expression and function of the trehalase genesNTH1 andYBR0106 inSaccharomyces cerevisiae. J Biol Chem 270:10193–10198Google Scholar
  43. Nwaka S, Mechler B, Destruelle M, Holzer H (1995b) Phenotypic features of trehalase mutants inSaccharomyces cerevisiae. FEBS Lett 360:286–290Google Scholar
  44. Panek AD (1991) Storage carbohydrates. In: Rose AH, Harrison JS (eds) The yeasts. Academic Press, New York, pp 655–678Google Scholar
  45. Panek AD, Mattoon JR (1977) Regulation of energy metabolism inSaccharomyces cerevisiae. Relationship between carbolite repression, trehalose synthesis and mitochondrial development. Arch Biochem Biophys 183:306–316Google Scholar
  46. Panek AD, Panek AC (1990) Metabolism and thermotolerance function of trehalose inSaccharomyces cerevisiae: a current perspective. J Biotechnol 14:229–238Google Scholar
  47. Panek AC, de Araujo PS, Neto VK, Panek AD (1987) Regulation of the trehalose-6-phosphate synthase complex inSaccharomyces cerevisiae. Curr Genet 11:459–465Google Scholar
  48. Panek AC, de Araujo PS, Poppe SC, Panek AD (1990) On the determination of trehalose-6-phosphate synthase inSaccharomyces cerevisiae. Biochem Int 21:695–704Google Scholar
  49. Pernambuco MB, Winderickx J, Crauwels M, Griffioen G, Mager WH, Thevelein JM (1986) Differential requirement for sugar phosphorylation in cells of the yeastSaccharomyces cerevisiae grown on glucose or grown on non-fermentable carbon sources for glucose-triggered signalling phenomena. Microbiology 142:1775–1782Google Scholar
  50. Rose MD, Winston F, Hieter P (1990) Methods in yeast genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  51. Schüller C, Brewster JL, Alexander MR, Gustin MC, Ruis H (1994) The HOG pathway controls osmotic regulation of transcription via stress response element (STRE) of theSaccharomyces cerevisiae CTT1 gene. EMBO J 13:4382–4389Google Scholar
  52. Singh KK, Norton RS (1991) Metabolic changes induced during adaptation ofSaccharomyces cerevisiae to a water stress. Arch Microbiol 165:38–42Google Scholar
  53. Sorger PK, Pelham HRB (1988) Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell 54:855–864Google Scholar
  54. Sur IP, Lobo Z, Maitra PK (1994) Analysis ofPFK3 — a gene involved in particulate phosphofructokinase synthesis reveals additional functions ofTPS2 inSaccharomyces cerevisiae. Yeast 10:199–209Google Scholar
  55. Thevelein JM (1984) Regulation of trehalose mobilisation in fungi. Microbiol Rev 48:42–59Google Scholar
  56. Thevelein JM (1988) Regulation of trehalose activity by phosphorylation-dephosphorylation during developmental transitions in fungi. Exp Mycol 12:1–12Google Scholar
  57. Thevelein JM (1991) Fermentable sugars and intracellular acidification as specific activators of the Ras adenylate cyclase signalling pathway in yeast. The relationship to nutrient-induced cell cycle control. Mol Microbiol 5:1301–1307Google Scholar
  58. Thevelein JM (1994) Signal transduction in yeast. Yeast 10:1753–1790Google Scholar
  59. Thevelein JM, Beullens M (1985) Cyclic AMP and the stimulation of trehalase activity in the yeastSaccharomyces cerevisiae by carbon sources, nitrogen sources and inhibitors of protein synthesis. J Gen Microbiol 131:3199–3209Google Scholar
  60. Thevelein JM, Hohmann S (1995) Trehalose synthase: guard to the gate of glycolysis in yeast? Trends Biochem Sci 20:3–10Google Scholar
  61. Thomas BJ, Rothstein RJ (1989) Elevated recombination rates in transcriptionally active DNA. Cell 56:619–630Google Scholar
  62. Toda T, Uno I, Ishikawa T, Powers S, Kataoka T, Broek D, Cameron S, Broach J, Matsumoto K, Wigler M (1985) in yeast, Ras proteins are controlling elements of adenylate cyclase. Cell 40:27–36Google Scholar
  63. Toda T, Cameron S, Sass P, Zoller M, Scott JD, McBullen B, Hurwitz M, Krebs EG, Wigler M (1987a) Cloning and characterization ofBCY1, a locus encoding a regulatory subunit of the cyclic AMP-dependent protein kinase inSaccharomyces cerevisiae. Mol Cell Biol 7:1371–1377Google Scholar
  64. Toda T, Cameron S, Sass P, Zoller M, Wigler M (1987b) Three different genes inSaccharomyces cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase. Cell 50:277–287Google Scholar
  65. Uno I, Matsumoto K, Adachi K, Ishikawa T (1983) Genetic and biochemical evidence that trehalase is a substrate of cAMP-dependent protein kinase in yeast. J Biol Chem 258:10867–10872Google Scholar
  66. Van Aelst L, Hohmann S, Bulaya B, De Koning W, Sierkstra L, Neves MJ, Luyten K, Alijo R, Ramos J, Coccetti P, Martegani E, de Magelhaes-Rocha NM, Brandao RL, Van Dijck P, Van Halewyn M, Dumez P, Jans AWH, Thevelein JM (1993) Molecular cloning of a gene involved in glucose sensing in the yeastSaccharomyces cerevisiae. Mol Microbiol 8:927–943Google Scholar
  67. Vandercammen A, François J, Hers H-G (1989) Characterisation of trehalose-6-phosphate synthetase and trehalose-6-phosphate phosphatase ofSaccharomyces cerevisiae. Eur J Biochem 182:613–620Google Scholar
  68. Van Dijck P, Colavizza D, Smet P, Thevelein JM (1995) Differential importance of trehalose in stress resistance in fermenting and nonfermentingSaccharomyces cerevisiae cells (1995) Appl Environ Microbiol 61:109–115Google Scholar
  69. Varela JCS, van Beekvelt C, Planta RJ, Mager WH (1992) Osmostress-induced changes in yeast gene expression. Mol Microbiol 6:2183–2190Google Scholar
  70. Varela JCS, Praekelt UM, Meacock PA, Planta RJ, Mager WH (1995) TheSaccharomyces cerevisiae HSP12 gene is activated by the high-osmolarity glycerol pathway and negatively regulated by protein kinase A. Mol Cell Biol 15:6232–6245Google Scholar
  71. Vuorio OE, Kalkkinen N, Londesborough J (1993) Cloning of two related genes encoding the 56-kDa and 123-kDa subunits of trehalose synthase from the yeastSaccharomyces cerevisiae. Eur J Biochem 216:848–861Google Scholar
  72. Wiemken A (1990) Trehalose in yeast: stress protectant rather than reserve carbohydrate. Antonie van Leeuwenhoek 58:209–217Google Scholar
  73. Winderickx J, Hino A, Crauwels M, Cobbaert P, Thevelein JM (1994) Characterisation of the ‘fermentable growth medium induced pathway’ inSaccharomyces cerevisiae. Arch Int Physiol Bioch Biophys 102:B44Google Scholar
  74. Wu A, Wemmie JA, Edgington NP, Goebl M, Guevara JL, Scott-Rowley W (1993) Yeast bZip proteins mediate pleiotropic drug and metal resistance. J Biol Chem 268:18850–18858Google Scholar
  75. Zamenhoff S (1957) Preparation and assay of deoxyribonucleic acids from animal tissues. Methods Enzymol 3:696–704Google Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • J. Winderickx
    • 1
  • J. H. de Winde
    • 1
  • M. Crauwels
    • 1
  • A. Hino
    • 1
  • S. Hohmann
    • 1
  • P. Van Dijck
    • 1
  • J. M. Thevelein
    • 1
  1. 1.Laboratorium voor Moleculaire CelbiologieKatholieke Universiteit LeuvenLeuven-HeverleeBelgium
  2. 2.National Food Research InstituteTsukubaJapan
  3. 3.Department of General and Marine MicrobiologyGöteborg UniversityGöteborgSweden
  4. 4.Janssen Research FoundationBeerseBelgium

Personalised recommendations