Current Genetics

, Volume 54, Issue 2, pp 71–81 | Cite as

Pseudohyphal differentiation defect due to mutations in GPCR and ammonium signaling is suppressed by low glucose concentration: a possible integrated role for carbon and nitrogen limitation

  • Revathi S. Iyer
  • Maitreyi Das
  • Paike Jayadeva Bhat
Research Article

Abstract

In response to carbon and/or nitrogen limitation, diploid cells of Saccharomyces cerevisiae either sporulate or develop pseudohyphae. Although the signal transduction pathways leading to these developmental changes have been extensively studied, how nutritional signals are integrated is not clearly understood. Results of this study indicate that reducing glucose concentration from 2% (SLAD) to 0.05% (SLALD) causes an increase in the magnitude of filamentation as well as a discernible reduction in the time required for pseudohyphal development. Further, the pseudohyphal defect of gpa2, gpr1and gpa2gpr1 but not the mep2 mutant strain is overcome on SLALD. Low glucose also induced pseudohyphae in mep2gpr1 but not mep2gpa2 strain suggesting that GPR1 inhibits pseudohyphae by inhibiting GPA2 function. Accordingly, deleting GPA2 in mep2gpr1 mutant abrogated pseudohyphae formation in SLALD. Further, replenishment of glucose suppressed pseudohyphal differentiation in wild-type cells grown in SLAD medium. However, in SLALD, glucose replenishment suppressed the filamentation response of gpa2 mutants but not that of strains carrying the wild-type GPA2. Increased trehalose levels correlated with decreased pseudohyphae formation. Results of this study demonstrate that filamentation in response to nitrogen limitation occurs as glucose becomes limiting.

Keywords

Yeast Glucose Pseudohyphae GPCR MEP2 Rapamycin 

Supplementary material

294_2008_202_MOESM1_ESM.doc (19 mb)
[Supplementary Figures] (DOC 19,431 kb)

References

  1. Adams BG (1972) Induction of galactokinase in Saccharomyces cerevisiae: kinetics of induction and glucose effects. J Bacteriol 111:308–315PubMedGoogle Scholar
  2. Adams A, Gottschling DE, Kaiser CA, Stearns T (1997) Methods in yeast genetics. Cold Spring Harbor Laboratory, New York, pp 1–157Google Scholar
  3. Beck T, Hall MN (1999) The TOR signaling pathway controls nuclear localization of nutrient-regulated transcription factors. Nature 402:689–692PubMedCrossRefGoogle Scholar
  4. Betram PG, Choi JH, Carvalho J, Chan T, Ai W, Steven Zheng XF (2002) Convergence of TOR-nitrogen and Snf1-glucose signaling pathways onto Gln3. Mol Cell Biol 22:1246–1252CrossRefGoogle Scholar
  5. Boeckstaens M, Andre B, Marini AM (2007) The yeast ammonium transport protein Mep2 and its positive regulator, the Npr1 kinase, play an important role in normal and pseudohyphal growth on various nitrogen media through retrieval of excreted ammonium. Mol Microbiol 64:534–546PubMedCrossRefGoogle Scholar
  6. Carlson M (1999) Glucose repression in yeast. Curr Opin Microbiol 2:202–207PubMedCrossRefGoogle Scholar
  7. Carlson M, Osmond BC, Neigeborn L, Botstein D (1984) A suppressor of SNF1 mutations causes constitutive high-level invertase synthesis yeast. Genetics 107:19–32PubMedGoogle Scholar
  8. Chen XJ, Clark-Walker GD (1999) The petite mutation in yeasts: 50 years on. Int Rev Cytol 194:197–237CrossRefGoogle Scholar
  9. Crespo JL, Hall MN (2002) Elucidating TOR signaling and rapamycin action: lessons from Saccharomyces cerevisiae. Microbiol Mol Bio Rev 66:579–591CrossRefGoogle Scholar
  10. Cutler NS, Pan X, Heitman J, Cardenas M (2001) The TOR signal transduction cascade controls cellular differentiation in response to nutrients. Mol Biol Cell 12:4103–4113PubMedGoogle Scholar
  11. Donzeau M, Bandlow W (1999) The yeast trimeric guanine nucleotide-binding protein α subunit, Gpa2p, controls the meiosis-specific kinase Ime2p activity in response to nutrients. Mol Cell Biol 19:6110–6119PubMedGoogle Scholar
  12. Engebrecht J (2003) Cell signaling in yeast sporulation. BBRC 306:325–328PubMedGoogle Scholar
  13. Erdman S, Snyder M (2001) A filamentous growth response mediated by the yeast mating pathway. Genetics 159:919–928PubMedGoogle Scholar
  14. Gancedo JM (2001) Control of pseudohyphae formation in Saccharomyces cerevisiae. FEMS Microbiol Rev 25:107–123PubMedCrossRefGoogle Scholar
  15. Gasch AP (2003) The environmental stress response: a common yeast response to diverse environmental stress. In: Hohmann S, Mager WH (eds) Yeast stress response. Springer-Verlag, Berlin, pp 11–57CrossRefGoogle Scholar
  16. Gimeno CJ, Ljungdahl PO, Styles CA, Fink GR (1992) Unipolar cell divisions in the yeast S. cerevisiae lead to filamentous growth: regulation by starvation and RAS. Cell 68:1077–1090PubMedCrossRefGoogle Scholar
  17. Gray JV, Petsko GA, Johnston GC, Ringe D, Singer RA, Werner-Wahsburne M (2004) “Sleeping beauty”: quiescence in Saccharomyces cerevisiae. Microbiol Mol Biol Rev 68:187–206PubMedCrossRefGoogle Scholar
  18. Harashima T, Heitman J (2002) The Gα protein Gpa2 controls yeast differentiation by interacting with kelch repeat proteins that mimic Gβ subunits. Mol Cell 10:163–173PubMedCrossRefGoogle Scholar
  19. Hedbacker K, Townley R, Carlson M (2004) Cyclic AMP-dependent protein kinase regulates the subcellular localization of Snf1-Sip1 protien kinase. Mol Cell Biol 24:1836–1843PubMedCrossRefGoogle Scholar
  20. Holstege FC, Jennings EG, Wyrick JJ, Lee TI, Hengartner CJ, Green MR, Golub TR, Lander ES, Young RA (1998) Dissecting the regulatory circuitry of a eukaryotic genome. Cell 95:717–728PubMedCrossRefGoogle Scholar
  21. Honigberg SM, Purnapatre K (2003) Signal pathway integration in the switch from the mitotic cell cycle to meiosis in yeast. J Cell Sci 116:2137–2147PubMedCrossRefGoogle Scholar
  22. Kienle I, Burgert M, Holzer H (1993) Assay of trehalose with acid trehalase purified from Saccharomyces cerevisiae. Yeast 9:607–611PubMedCrossRefGoogle Scholar
  23. Kubler E, Mosch H, Rupp S, Lisanti MP (1997) Gpa2p, a G-protein α-subunit, regulates growth and pseudohyphal development in Saccharomyces cerevsiae via a cAMP-dependent mechanism. J Biol Chem 272:20321–20323PubMedCrossRefGoogle Scholar
  24. Kuchin S, Vyas VK, Carlson M (2002) Snf1 protein kinase and the repressors Nrg1 and Nrg2 regulate FLO11, haploid invasive growth and diploid pseudohyphal growth. Mol Cell Biol 22:3994–4000PubMedCrossRefGoogle Scholar
  25. Lengeler KB, Davidson RC, D’Souza C, Harashima T, Shen W, Wang P, Pan X, Waugh M, Heitman J (2000) Signal transduction cascades regulating fungal development and virulence. Microbiol Mol Biol Rev 64:746–785PubMedCrossRefGoogle Scholar
  26. Lo W, Dranginis AM (1998) The cell surface flocculin Flo11 is required for pseudohyphae formation and invasion by Saccharomyces cerevisiae. Mol Biol Cell 9:161–171PubMedGoogle Scholar
  27. Lorenz MC, Heitman J (1997) Yeast pseudohyphal growth is regulated by GPA2, a G protein α homolog. EMBO J 16:7008–7018PubMedCrossRefGoogle Scholar
  28. Lorenz MC, Heitman J (1998) The MEP2 ammonium permease regulates pseudohyphal differentiation Saccharomyces cerevisiae. EMBO J 17:1236–1247PubMedCrossRefGoogle Scholar
  29. Lorenz MC, Pan X, Harashima T, Cardenas ME, Xue Y, Hirsch JP, Heitman J (2000) The G protein-coupled receptor Gpr1 is a nutrient sensor that regulates pseudohyphal differentiation in Saccharomyces cerevisiae. Genetics 154:609–622PubMedGoogle Scholar
  30. Lu A, Hirsch JP (2005) Cyclic AMP-independent regulation of protein kinase: a substrate phosphorylation by Kelch repeat homologues. Eukaryot Cell 4:1794–1800PubMedCrossRefGoogle Scholar
  31. Marini A, Soussi-Boudekou S, Vissers S, Andre B (1997) A family of ammonium transporters in Saccharomyces cerevisiae. Mol Cell Biol 17:4282–4283PubMedGoogle Scholar
  32. Mosch H, Roberts RL, Fink G (1996) Ras2 signals via the Cdc42/Ste20/mitogen-activated protein kinase module to induce filamenatous growth in Saccharomyces cerevisiae. Proc Natl Acad Sci USA 93:5352–5356PubMedCrossRefGoogle Scholar
  33. Orlova M, Kanter E, Krakovich D, Kuchin S (2006) Nitrogen availability and TOR regulate the Snf1 protein kinase in Saccharomyces cerevisiae. Eukaryot Cell 5:1831–1837PubMedCrossRefGoogle Scholar
  34. Palecek SP, Parikh AS, Kron SJ (2002) Sensing, signaling and integrating physical processes during Saccharomyces cerevisiae invasive and filamentous growth. Microbiol 148:893–907Google Scholar
  35. Pan X, Heitman J (2002) Protein kinase A operates a molecular switch that governs yeast pseudohyphal differentiation. Mol Cell Biol 22:3981–3993PubMedCrossRefGoogle Scholar
  36. Peeters T, Louwet W, Gelade R, Nauwelaers D, Thevelein JM, Versele M (2006) Kelch-repeat proteins interacting with the G{alpha} protein Gpa2 bypass adenylate cyclase for direct regulation of protein kinase A in yeast. Proc Natl Acad Sci USA 103:13034–13039PubMedCrossRefGoogle Scholar
  37. Piskur J, Rozpedowska E, Polakava S, Merico A, Compagno A (2006) How did Saccharomyces cerevisiae become a good brewer? Trends Genet 22:183–187PubMedCrossRefGoogle Scholar
  38. Roberts RL, Fink GR (1994) Elements of a single MAP kinase cascade in Saccharomyces cerevisiae mediate two developmental programs in the same cell type: mating and invasive growth. Genes Dev 8:2974–2985PubMedCrossRefGoogle Scholar
  39. Rupp S, Summers E, Lo HJ, Madhani H, Fink G (1999) MAP kinase and cAMP filamentation signaling pathways converge on the unusually large promoter of the yeast FLO11 gene. EMBO J 18:1257–1269PubMedCrossRefGoogle Scholar
  40. Thevelein JM, Gelade R, Holsbeeks I, Legatie O, Popova Y, Rolland F, Stolz F, Van de Velde S, Van Dijck P et al (2005) Nutrient sensing systems for rapid activation of the protein kinase A pathway in yeast. Biochem Soc Trans 33:253–256PubMedCrossRefGoogle Scholar
  41. Thomas G, Hall MN (1997) TOR signaling and control of cell growth. Curr Opin Cell Biol 9:782–787PubMedCrossRefGoogle Scholar
  42. Van de Velde S, Thevelein JM (2008) cAMP-PKA and Snf1 signaling mechanisms underlie the superior potency of sucrose for induction of filamentation in yeast. Eukaryot Cell 7:286–293PubMedCrossRefGoogle Scholar
  43. van Nuland A, Vandormael P, Donaton M, Alenquer M, Lourenco A, Quintino E, Versele M, Thevelein JM (2006) Ammonium permease-based sensing mechanism for rapid ammonium activation of the protein kinase: a pathway in yeast. Mol Microbiol 59:1485–1505PubMedCrossRefGoogle Scholar
  44. Xie MW, Jin F, Hwang H, Hwang S, Anand V, Duncan MC, Huang J (2005) Insights into TOR function and rapamycin response: chemical genomic profiling by using a high-density cell array method. Proc Natl Acad Sci USA 102:7215–7220PubMedCrossRefGoogle Scholar
  45. Xue Y, Batlle M, Hirsch JP (1998) GPR1 encodes a putative G protein-coupled receptor that associates with the Gpa2 Gα subunit and functions in a Ras-independent pathway. EMBO J 17:1996–2007PubMedCrossRefGoogle Scholar
  46. Zurita-Martinez SA, Cardenas ME (2005) Tor and cyclic AMP-Protein kinase A: two parallel pathways regulating expression of genes required for cell growth. Eukaryot Cell 4:63–71PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Revathi S. Iyer
    • 1
  • Maitreyi Das
    • 2
  • Paike Jayadeva Bhat
    • 1
  1. 1.Laboratory of Molecular GeneticsSchool of Biosciences and Bioengineering, Indian Institute of TechnologyMumbaiIndia
  2. 2.Department of Molecular and Cellular PharmacologyUniversity of Miami Miller School of MedicineMiamiUSA

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