Advertisement

Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Mutations in GCR1 affect SUC2 gene expression in Saccharomyces cerevisiae

Abstract

Transcription of SUC2, the gene that encodes the cytoplasmic and secreted forms of the enzyme invertase, is controlled by glucose repression and derepression mechanisms in Saccharomyces cerevisiae. Several regulatory factors such as the Mig1p-Tup1p-Ssn6p repressor complex and the Snf1p kinase complex have been identified previously as regulators of SUC2 expression. We show that, in addition to these factors, expression of SUC2 is affected by mutations in the gene GCR1 that encodes the glycolysis regulatory protein Gcr1p. Expression of Suc2-LacZ was not repressed by glucose in gcr1 mutant yeast cells exposed to glucose. Furthermore, secreted invertase activity was constitutively expressed under glucose-repressed and derepressed conditions in gcr1 mutants. DNA gel mobility shift assays and in-vitro DNase I protection experiments mapped a DNA binding site for Gcr1p in the transcriptional control region of the SUC2 gene, next to a previously mapped Mig1p binding site. However, the mechanism by which gcr1 mutations relieve glucose repression remains obscure.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2

References

  1. Ausubel SM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1993) Current protocols in molecular biology. Greene Publishing Associates and Wiley Interscience. NewYork

  2. Baker HV (1991) GCR1of Saccharomyces cerevisiae encodes a DNA binding protein whose binding is abolished by mutations in the CTTCC sequence motif. Proc Natl Acad Sci USA 88:9443–9447

  3. Bisson LF, Coons DM, Kruckeberg AL, Lewis DA (1993) Yeast sugar transporters. Crit Rev Biochem Mol Biol 28:259–308

  4. Boles E, Hollenberg CP (1997) The molecular genetics of hexose transport in yeasts. FEMS Microbiol Rev 21:85–111

  5. Boles E, Müller S, Zimmermann FK (1996) A multi-layered sensory system controls yeast glycolytic gene expression. Mol Microbiol 19:641–642

  6. Bu Y, Schmidt MC (1998) Identification of cis -acting elements in the SUC2promoter of Saccharomyces cerevisiae required for activation of transcription. Nucleic Acids Res 26:1002–1009

  7. Carlson M (1998) Regulation of glucose utilization in yeast. Curr Opin Genet Dev 8:560–564

  8. Carlson M, Botstein D (1982) Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase. Cell 28:145–154

  9. Clifton D, Fraenkel DG (1981) The gcr (glycolysis regulation) mutation of Saccharomyces cerevisiae. J Biol Chem 256:13074–13078

  10. DeVit MJ, Wadle JA, Johnson M (1997) Regulated nuclear translocation of Mig1 glucose repressor. Mol Biol Cell 8:1603–1618

  11. Drazinic CM, Smerage JB, López MC, Baker HV (1996) Activation mechanism of the multifunctional transcription factor repressor-activator protein 1 (Rap1p). Mol Cell Biol 16:3187–3196

  12. Dudley AM, Gansheroff LJ, Winston F (1999) Specific components of the SAGA complex are required for Gcn4- and Gcr1-mediated activation of the his4–912δ promoter in Saccharomyces cerevisiae. Genetics 151:1365–1378

  13. Edmondson DG, Smith MM, Roth SY (1996) Repression domain of the yeast global repressor TUP1 interacts directly with histones H3 and H4. Genes Dev 10:1247–1259

  14. Gancedo JM (1992) Carbon catabolite repression in yeast. Eur J Biochem 206:297–313

  15. Gavin IM, Simpson RT (1997) Interplay of yeast global transcriptional regulators Ssn6p-Tup1p and Swi-Snf and their effect on chromatin structure. EMBO J 16:6263–6271

  16. Goldstein A, Lampen JO (1975) β-D-Fructofuranoside fructohydrolase from yeast. Methods Enzymol 42:504–511

  17. Guarente L (1983) Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol 101:181–191

  18. Guarente L, Ptashne M (1981) Fusion of E. coli LacZ to cytochrome c gene of Saccharomyces cerevisiae. Proc Natl Acad Sci USA 78:2199–2203

  19. Guthrie C, Fink GR (1991) Guide to yeast genetics and molecular biology. Academic Press, San Diego

  20. Hardie DG, Carling D, Halford N (1994) Roles of the Snf1/Rkin1/AMP-activated protein kinase family in the response to environmental and nutritional stress. Semin Cell Biol 5:409–416

  21. Hardie DG, Carling D, Carlson M (1998) The AMP-activated/SNF1 protein kinase subfamily: metabolic sensors of the eukaryotic cell? Annu Rev Biochem 67:821–855

  22. Herrero P, Martinez-Campa C, Moreno F (1998) The hexokinase 2 protein participates in regulatory DNA-protein complexes necessary for glucose repression of the SUC2gene in Saccharomyces cerevisiae. FEBS Lett 434:71–76

  23. Higgins VJ, Braidwood M, Bell P, Bissinger P, Dawes IW, Attfield PV (1999) Genetic evidence that high noninduced maltase and maltose permease activities, governed by MALx3-encoded transcriptional regulators, determine efficiency of gas production by bakers yeast in unsugared dough. Appl Environ Microbiol 65:680–685

  24. Huie MA, Scott EW, Drazinic CM, López MC, Hornstra IK, Yang TP, Baker HV (1992) Characterization of the DNA-binding activity of Gcr1: in-vivo evidence for two GCR1-binding sites in the upstream activating sequence of TPI of Saccharomyces cerevisiae. Mol Cell Biol 12:2690–2700

  25. Ito H, Fukuda Y, Murata K, Kimura A (1983) Transformation of intact yeast cells treated with alkali cations. J Bacteriol 153:163–168

  26. Lenburg ME, O'Shea EK (2001) Genetic evidence for a morphogenic function of the Saccharomyces cerevisiae Pho85 cyclin-dependent kinase. Genetics 157:39–51

  27. López MC, Baker HV (2000) Understanding the growth phenotype of the yeast gcr1mutant in terms of global genomic expression patterns. J Bacteriol 182:4970–4978

  28. López MC, Smerage JB, Baker HV (1998) Multiple domains of repressor activator protein contribute to facilitated binding of glycolysis regulatory protein 1. Proc Natl Acad Sci USA 95:14112–14117

  29. Lundin M, Nehlin JO, Ronne H (1994) Importance of a flanking AT-rich region in target site recognition by the GC box-binding zinc finger protein MIG1. Mol Cell Biol 14:1979–1985

  30. Lutfiyya LL, Johnston M (1996) Two zinc-finger-containing repressors are responsible for glucose repression of SUC2expression. Mol Cell Biol 16:4790–4797

  31. Nagawa F, Fink GR (1985) The relationship between the "TATA" sequence and transcription initiation sites at the HIS4gene of Saccharomyces cerevisiae. Proc Natl Acad Sci USA 82:8557–8561

  32. Özcan S, Johnston M (1996) Two different repressors collaborate to restrict expression of the yeast glucose transporter genes HXT2and HXT4to low levels of glucose. Mol Cell Biol 16:5536–5545

  33. Palecek SP, Parikh AS, Kron SJ (2000) Genetic analysis reveals that FLO11upregulation and cell polarization independently regulate invasive growth in Saccharomyces cerevisiae. Genetics 156:1005–1023

  34. Pavlovic B, Hörz W (1988) The chromatin structure of the promoter of a glyceraldehyde phosphate dehydrogenase gene from Saccharomyces cerevisiae reflects its functional state. Mol Cell Biol 8:5513–5520

  35. Perlman D, Halvorson HO, Cannon LE (1982) Presecretory and cytoplasmic invertase polypeptides encoded by distinct mRNAs derived from the same structural gene differ by a signal sequence. Proc Natl Acad Sci USA 79:781–785

  36. Perlman D, Raney P, Halvorson HO (1984) Cytoplasmic and secreted Saccharomyces cerevisiae invertase mRNAs encoded by one gene can be differentially or coordinately regulated. Mol Cell Biol 9:1682–1688

  37. Ronne H (1995) Glucose repression in Fungi. Trends in Genet 11:12–17

  38. Rothe C, Lehle L (1998) Sorting of invertase signal peptide mutants in yeast dependent and independent on the signal recognition particle. Eur J Biochem 252:16–24

  39. Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual (2nd edn). Cold Spring Harbor Laboratory Press, Cold Spring Harbor. N.Y.

  40. Sarokin L, Carlson M (1985) Upstream region of the SUC2gene confers regulated expression to a heterologous gene in Saccharomyces cerevisiae. Mol Cell Biol 5: 2521–2526

  41. Scott EW, Baker HV (1993) Concerted action of the transcriptional activators REB1, RAP1 and GCR1 in the high-level expression of the glycolytic gene TPI. Mol Cell Biol 13:543–550

  42. Scott EW, Allison HE, Baker HV (1990) Characterization of TPI gene expression in isogenic wild-type and gcr1-deletion mutant strains of Saccharomyces cerevisiae. Nucleic Acids Res 18:7099–7107

  43. Stanway CA, Gibbs JM, Kearsey SE, Lopez MC, Baker HV (1994) The yeast co-activator Gal11 positively influences transcription of the phosphoglycerate kinase gene, but only when RAP1 is bound to its upstream activation sequence. Mol Gen Genet 243:207–214

  44. Sundarsanam P, Winston F (2000) The Swi/Snf family nucleosome-remodeling complexes and transcriptional control. Trends Genet 16: 345–351

  45. Trumbly RJ (1992) Glucose repression in the yeast Saccharomyces cerevisiae. Mol Microbiol 6:15–21

  46. Türkel S, Bisson LF (1999) Transcription of the HXT4gene is regulated by Gcr1p and Gcr2p in the yeast S. cerevisiae. Yeast 15:1047–1057

  47. Türkel S, Liao X-B, Farabaugh PJ (1997) GCR1-dependent transcriptional activation of yeast retrotransposon Ty2–917. Yeast 13:917–930

  48. Uemura H, Fraenkel DG (1990) gcr2, a new mutation affecting glycolytic gene expression in Saccharomyces cerevisiae. Mol Cell Biol 10:6389–6396

  49. Uemura H, Koshio M, Inoue Y, López MC, Baker HV (1997) The role of Gcr1p in the transcriptional activation of glycolytic genes in yeast Saccharomyces cerevisiae. Genetics 147:521–532

  50. Vallier LG, Carlson M (1991) New SNF genes GAL11and GRR1affect SUC2expression in Saccharomyces cerevisiae. Genetics 129:675–684

  51. Wu L, Winston F (1997) Evidence that Snf-Swi controls chromatin structure over both the TATA and UAS regions of the SUC2promoter in Saccharomyces cerevisiae. Nucleic Acids Res 25:4230–4234

Download references

Acknowledgements

We thank Dr. Akira Sakai of the Mitsubishi Chemical Corp., Japan for providing plasmids pZU and pZU-SUC2. Research in S. Türkel's laboratory was supported by The Scientific and Technical Research Council of Turkey (TUBITAK, TBAG-1979). Research conducted in H. V. Baker's lab was supported by a grant from the National Science Foundation (MCB9816990).

Author information

Correspondence to H. V. Baker.

Additional information

Communicated by C. P. Hollenberg

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Türkel, S., Turgut, T., López, M.C. et al. Mutations in GCR1 affect SUC2 gene expression in Saccharomyces cerevisiae . Mol Gen Genomics 268, 825–831 (2003). https://doi.org/10.1007/s00438-003-0808-4

Download citation

Keyword

  • Saccharomyces cerevisiae
  • SUC2
  • Invertase
  • GCR1
  • Glucose repression