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. 1995 May;15(5):2646–2653. doi: 10.1128/mcb.15.5.2646

The GCR1 requirement for yeast glycolytic gene expression is suppressed by dominant mutations in the SGC1 gene, which encodes a novel basic-helix-loop-helix protein.

K Nishi 1, C S Park 1, A E Pepper 1, G Eichinger 1, M A Innis 1, M J Holland 1
PMCID: PMC230494  PMID: 7739544

Abstract

The GCR1 gene product is required for maximal transcription of yeast glycolytic genes and for growth of yeast strains in media containing glucose as a carbon source. Dominant mutations in two genes, SGC1 and SGC2, as well as recessive mutations in the SGC5 gene were identified as suppressors of the growth and transcriptional defects caused by a gcr1 null mutation. The wild-type and mutant alleles of SGC1 were cloned and sequenced. The predicted amino acid sequence of the SGC1 gene product includes a region with substantial similarity to the basic-helix-loop-helix domain of the Myc family of DNA-binding proteins. The SGC1-1 dominant mutant allele contained a substitution of glutamine for a highly conserved glutamic acid residue within the putative basic DNA binding domain. A second dominant mutant, SGC1-2, contained a valine-for-isoleucine substitution within the putative loop region. The SGC1-1 dominant mutant suppressed the GCR1 requirement for enolase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, and pyruvate kinase gene expression. Expression of the yeast enolase genes was reduced three- to fivefold in strains carrying an sgc1 null mutation, demonstrating that SGC1 is required for maximal enolase gene expression. Expression of the enolase genes in strains carrying gcr1 and sgc1 double null mutations was substantially less than observed for strains carrying either null mutation alone, suggesting that GCR1 and SGC1 function on parallel pathways to activate yeast glycolytic gene expression.

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Selected References

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  1. Alber T., Kawasaki G. Nucleotide sequence of the triose phosphate isomerase gene of Saccharomyces cerevisiae. J Mol Appl Genet. 1982;1(5):419–434. [PubMed] [Google Scholar]
  2. Baker H. V. GCR1 of Saccharomyces cerevisiae encodes a DNA binding protein whose binding is abolished by mutations in the CTTCC sequence motif. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9443–9447. doi: 10.1073/pnas.88.21.9443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker H. V. Glycolytic gene expression in Saccharomyces cerevisiae: nucleotide sequence of GCR1, null mutants, and evidence for expression. Mol Cell Biol. 1986 Nov;6(11):3774–3784. doi: 10.1128/mcb.6.11.3774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berben G., Legrain M., Gilliquet V., Hilger F. The yeast regulatory gene PHO4 encodes a helix-loop-helix motif. Yeast. 1990 Sep-Oct;6(5):451–454. doi: 10.1002/yea.320060510. [DOI] [PubMed] [Google Scholar]
  5. Bitter G. A., Chang K. K., Egan K. M. A multi-component upstream activation sequence of the Saccharomyces cerevisiae glyceraldehyde-3-phosphate dehydrogenase gene promoter. Mol Gen Genet. 1991 Dec;231(1):22–32. doi: 10.1007/BF00293817. [DOI] [PubMed] [Google Scholar]
  6. Breeden L., Nasmyth K. Regulation of the yeast HO gene. Cold Spring Harb Symp Quant Biol. 1985;50:643–650. doi: 10.1101/sqb.1985.050.01.078. [DOI] [PubMed] [Google Scholar]
  7. Brindle P. K., Holland J. P., Willett C. E., Innis M. A., Holland M. J. Multiple factors bind the upstream activation sites of the yeast enolase genes ENO1 and ENO2: ABFI protein, like repressor activator protein RAP1, binds cis-acting sequences which modulate repression or activation of transcription. Mol Cell Biol. 1990 Sep;10(9):4872–4885. doi: 10.1128/mcb.10.9.4872. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buchman A. R., Lue N. F., Kornberg R. D. Connections between transcriptional activators, silencers, and telomeres as revealed by functional analysis of a yeast DNA-binding protein. Mol Cell Biol. 1988 Dec;8(12):5086–5099. doi: 10.1128/mcb.8.12.5086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Burke R. L., Tekamp-Olson P., Najarian R. The isolation, characterization, and sequence of the pyruvate kinase gene of Saccharomyces cerevisiae. J Biol Chem. 1983 Feb 25;258(4):2193–2201. [PubMed] [Google Scholar]
  10. Cai M., Davis R. W. Yeast centromere binding protein CBF1, of the helix-loop-helix protein family, is required for chromosome stability and methionine prototrophy. Cell. 1990 May 4;61(3):437–446. doi: 10.1016/0092-8674(90)90525-j. [DOI] [PubMed] [Google Scholar]
  11. Carmen A. A., Holland M. J. The upstream repression sequence from the yeast enolase gene ENO1 is a complex regulatory element that binds multiple trans-acting factors including REB1. J Biol Chem. 1994 Apr 1;269(13):9790–9797. [PubMed] [Google Scholar]
  12. Casadaban M. J., Martinez-Arias A., Shapira S. K., Chou J. Beta-galactosidase gene fusions for analyzing gene expression in escherichia coli and yeast. Methods Enzymol. 1983;100:293–308. doi: 10.1016/0076-6879(83)00063-4. [DOI] [PubMed] [Google Scholar]
  13. Chambers A., Stanway C., Tsang J. S., Henry Y., Kingsman A. J., Kingsman S. M. ARS binding factor 1 binds adjacent to RAP1 at the UASs of the yeast glycolytic genes PGK and PYK1. Nucleic Acids Res. 1990 Sep 25;18(18):5393–5399. doi: 10.1093/nar/18.18.5393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Clifton D., Fraenkel D. G. The gcr (glycolysis regulation) mutation of Saccharomyces cerevisiae. J Biol Chem. 1981 Dec 25;256(24):13074–13078. [PubMed] [Google Scholar]
  15. Cohen R., Holland J. P., Yokoi T., Holland M. J. Identification of a regulatory region that mediates glucose-dependent induction of the Saccharomyces cerevisiae enolase gene ENO2. Mol Cell Biol. 1986 Jul;6(7):2287–2297. doi: 10.1128/mcb.6.7.2287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Cohen R., Yokoi T., Holland J. P., Pepper A. E., Holland M. J. Transcription of the constitutively expressed yeast enolase gene ENO1 is mediated by positive and negative cis-acting regulatory sequences. Mol Cell Biol. 1987 Aug;7(8):2753–2761. doi: 10.1128/mcb.7.8.2753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ferré-D'Amaré A. R., Prendergast G. C., Ziff E. B., Burley S. K. Recognition by Max of its cognate DNA through a dimeric b/HLH/Z domain. Nature. 1993 May 6;363(6424):38–45. doi: 10.1038/363038a0. [DOI] [PubMed] [Google Scholar]
  18. Fisher F., Goding C. R. Single amino acid substitutions alter helix-loop-helix protein specificity for bases flanking the core CANNTG motif. EMBO J. 1992 Nov;11(11):4103–4109. doi: 10.1002/j.1460-2075.1992.tb05503.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gallwitz D., Seidel R. Molecular cloning of the actin gene from yeast Saccharomyces cerevisiae. Nucleic Acids Res. 1980 Mar 11;8(5):1043–1059. doi: 10.1093/nar/8.5.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hitzeman R. A., Hagie F. E., Hayflick J. S., Chen C. Y., Seeburg P. H., Derynck R. The primary structure of the Saccharomyces cerevisiae gene for 3-phosphoglycerate kinase. Nucleic Acids Res. 1982 Dec 11;10(23):7791–7808. doi: 10.1093/nar/10.23.7791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Holland J. P., Holland M. J. The primary structure of a glyceraldehyde-3-phosphate dehydrogenase gene from Saccharomyces cerevisiae. J Biol Chem. 1979 Oct 10;254(19):9839–9845. [PubMed] [Google Scholar]
  22. Holland J. P., Labieniec L., Swimmer C., Holland M. J. Homologous nucleotide sequences at the 5' termini of messenger RNAs synthesized from the yeast enolase and glyceraldehyde-3-phosphate dehydrogenase gene families. The primary structure of a third yeast glyceraldehyde-3-phosphate dehydrogenase gene. J Biol Chem. 1983 Apr 25;258(8):5291–5299. [PubMed] [Google Scholar]
  23. Holland M. J., Yokoi T., Holland J. P., Myambo K., Innis M. A. The GCR1 gene encodes a positive transcriptional regulator of the enolase and glyceraldehyde-3-phosphate dehydrogenase gene families in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Feb;7(2):813–820. doi: 10.1128/mcb.7.2.813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Huie M. A., Scott E. W., Drazinic C. M., Lopez M. C., Hornstra I. K., Yang T. P., Baker H. V. 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. 1992 Jun;12(6):2690–2700. doi: 10.1128/mcb.12.6.2690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kohalmi S. E., Kunz B. A. Role of neighbouring bases and assessment of strand specificity in ethylmethanesulphonate and N-methyl-N'-nitro-N-nitrosoguanidine mutagenesis in the SUP4-o gene of Saccharomyces cerevisiae. J Mol Biol. 1988 Dec 5;204(3):561–568. doi: 10.1016/0022-2836(88)90355-5. [DOI] [PubMed] [Google Scholar]
  27. McAlister L., Holland M. J. Differential expression of the three yeast glyceraldehyde-3-phosphate dehydrogenase genes. J Biol Chem. 1985 Dec 5;260(28):15019–15027. [PubMed] [Google Scholar]
  28. Nikoloff D. M., McGraw P., Henry S. A. The INO2 gene of Saccharomyces cerevisiae encodes a helix-loop-helix protein that is required for activation of phospholipid synthesis. Nucleic Acids Res. 1992 Jun 25;20(12):3253–3253. doi: 10.1093/nar/20.12.3253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rodicio R., Heinisch J. J., Hollenberg C. P. Transcriptional control of yeast phosphoglycerate mutase-encoding gene. Gene. 1993 Mar 30;125(2):125–133. doi: 10.1016/0378-1119(93)90319-x. [DOI] [PubMed] [Google Scholar]
  30. Scott E. W., Baker H. V. Concerted action of the transcriptional activators REB1, RAP1, and GCR1 in the high-level expression of the glycolytic gene TPI. Mol Cell Biol. 1993 Jan;13(1):543–550. doi: 10.1128/mcb.13.1.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Slater M. R., Craig E. A. Transcriptional regulation of an hsp70 heat shock gene in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1987 May;7(5):1906–1916. doi: 10.1128/mcb.7.5.1906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Uemura H., Fraenkel D. G. gcr2, a new mutation affecting glycolytic gene expression in Saccharomyces cerevisiae. Mol Cell Biol. 1990 Dec;10(12):6389–6396. doi: 10.1128/mcb.10.12.6389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Uemura H., Jigami Y. Role of GCR2 in transcriptional activation of yeast glycolytic genes. Mol Cell Biol. 1992 Sep;12(9):3834–3842. doi: 10.1128/mcb.12.9.3834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Voronova A., Baltimore D. Mutations that disrupt DNA binding and dimer formation in the E47 helix-loop-helix protein map to distinct domains. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4722–4726. doi: 10.1073/pnas.87.12.4722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Willett C. E., Gelfman C. M., Holland M. J. A complex regulatory element from the yeast gene ENO2 modulates GCR1-dependent transcriptional activation. Mol Cell Biol. 1993 Apr;13(4):2623–2633. doi: 10.1128/mcb.13.4.2623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yoshida K., Ogawa N., Oshima Y. Function of the PHO regulatory genes for repressible acid phosphatase synthesis in Saccharomyces cerevisiae. Mol Gen Genet. 1989 May;217(1):40–46. doi: 10.1007/BF00330940. [DOI] [PubMed] [Google Scholar]

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