Home LiteratureArticle Details
PMID: 1682801 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Role of the complex upstream region of the GDH2 gene in nitrogen regulation of the NAD-linked glutamate dehydrogenase in Saccharomyces cerevisiae.

Molecular and cellular biology ·Vol. 11 ·No. 12 ·1991-12-00 ·Pages 6229-47

Miller SM, Magasanik B

Abstract

We analyzed the upstream region of the GDH2 gene, which encodes the NAD-linked glutamate dehydrogenase in Saccharomyces cerevisiae, for elements important for the regulation of the gene by the nitrogen source. The levels of this enzyme are high in cells grown with glutamate as the sole source of nitrogen and low in cells grown with glutamine or ammonium. We found that this regulation occurs at the level of transcription and that a total of six sites are required to cause a CYC1-lacZ fusion to the GDH2 gene to be regulated in the same manner as the NAD-linked glutamate dehydrogenase. Two sites behaved as upstream activation sites (UASs). The remaining four sites were found to block the effects of the two UASs in such a way that the GDH2-CYC1-lacZ fusion was not expressed unless the cells containing it were grown under conditions favorable for the activity of both UASs. This complex regulatory system appears to account for the fact that GDH2 expression is exquisitely sensitive to glutamine, whereas the expression of GLN1, coding for glutamine synthetase, is not nearly as sensitive.

Related Genes
GDH2
MeSH Terms
Amino Acid Sequence Ammonia/metabolism Base Sequence Cloning, Molecular DNA, Fungal Gene Expression Regulation, Fungal Glutamate Dehydrogenase/genetics,metabolism Glutamates/metabolism Glutamic Acid Molecular Sequence Data Nitrogen/metabolism Protein Biosynthesis RNA, Fungal/metabolism RNA, Messenger/metabolism Regulatory Sequences, Nucleic Acid Restriction Mapping Saccharomyces cerevisiae/enzymology,genetics Transcription, Genetic
Chemicals
DNA, Fungal Glutamates RNA, Fungal RNA, Messenger Glutamic Acid Ammonia Glutamate Dehydrogenase Nitrogen
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Miller S M
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
Magasanik B
References (58)
58 references, click to expand
  1. Activation of yeast polymerase II transcription by herpesvirus VP16 and GAL4 derivatives in vitro.
    Mol Cell Biol. 1989 Nov;9(11):4746-9 PMID: 2557540
  2. Possible failure of NADP-glutamate dehydrogenase to participate directly in nitrogen repression of the allantoin degradative enzymes in Saccharomyces cerevisiae.
    Biochem Biophys Res Commun. 1975 Oct 6;66(3):889-92 PMID: 241348
  3. THE PATHWAY OF ARGININE BREAKDOWN IN SACCHAROMYCES CEREVISIAE.
    Biochim Biophys Acta. 1964 Dec 9;93:650-2 PMID: 14263163
  4. The URE2 gene product of Saccharomyces cerevisiae plays an important role in the cellular response to the nitrogen source and has homology to glutathione s-transferases.
    Mol Cell Biol. 1991 Feb;11(2):822-32 PMID: 1990286
  5. Role of integration host factor in the regulation of the glnHp2 promoter of Escherichia coli.
    Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1631-5 PMID: 2000372
  6. Sequence and expression of GLN3, a positive nitrogen regulatory gene of Saccharomyces cerevisiae encoding a protein with a putative zinc finger DNA-binding domain.
    Mol Cell Biol. 1991 Dec;11(12):6216-28 PMID: 1682800
  7. Physiological and genetic analysis of the carbon regulation of the NAD-dependent glutamate dehydrogenase of Saccharomyces cerevisiae.
    Mol Cell Biol. 1991 Sep;11(9):4455-65 PMID: 1652057
  8. Site-directed mutagenesis of the 'zinc finger' DNA-binding domain of the nitrogen-regulatory protein NIT2 of Neurospora.
    Mol Microbiol. 1990 Nov;4(11):1847-52 PMID: 2150539
  9. Requirement of upstream activation sequences for nitrogen catabolite repression of the allantoin system genes in Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Dec;9(12):5440-4 PMID: 2511434
  10. Identification of sequences responsible for transcriptional activation of the allantoate permease gene in Saccharomyces cerevisiae.
    Mol Cell Biol. 1989 Feb;9(2):602-8 PMID: 2651902
  11. The erythroid-specific transcription factor Eryf1: a new finger protein.
    Cell. 1989 Sep 8;58(5):877-85 PMID: 2776214
  12. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells.
    Nature. 1989 Jun 8;339(6224):446-51 PMID: 2725678
  13. Regulation of nitrogen assimilation in Saccharomyces cerevisiae: roles of the URE2 and GLN3 genes.
    J Bacteriol. 1988 Feb;170(2):708-13 PMID: 2892826
  14. Regulation of glutamine-repressible gene products by the GLN3 function in Saccharomyces cerevisiae.
    Mol Cell Biol. 1984 Dec;4(12):2758-66 PMID: 6152012
  15. Adenovirus-2 E1A products repress enhancer-induced stimulation of transcription.
    Nature. 1984 Dec 13-19;312(5995):608-12 PMID: 6095113
  16. A bacterial repressor protein or a yeast transcriptional terminator can block upstream activation of a yeast gene.
    Nature. 1984 Dec 13-19;312(5995):612-5 PMID: 6390216
  17. Molecular analysis of SSN6, a gene functionally related to the SNF1 protein kinase of Saccharomyces cerevisiae.
    Mol Cell Biol. 1987 Oct;7(10):3637-45 PMID: 3316983
  18. Transcriptional regulation of the DAL5 gene in Saccharomyces cerevisiae.
    J Bacteriol. 1987 Aug;169(8):3521-4 PMID: 3301804
  19. Ubiquitous upstream repression sequences control activation of the inducible arginase gene in yeast.
    Proc Natl Acad Sci U S A. 1987 Jun;84(12):3997-4001 PMID: 3295874
  20. Identification of the ureidoglycolate hydrolase gene in the DAL gene cluster of Saccharomyces cerevisiae.
    Mol Cell Biol. 1985 Sep;5(9):2279-88 PMID: 3915539
  21. Specific DNA binding of GAL4, a positive regulatory protein of yeast.
    Cell. 1985 Apr;40(4):767-74 PMID: 3886158
  22. The GLN3 gene product is required for transcriptional activation of allantoin system gene expression in Saccharomyces cerevisiae.
    J Bacteriol. 1990 Feb;172(2):1014-8 PMID: 2153652
  23. A cis-acting element present in multiple genes serves as a repressor protein binding site for the yeast CAR1 gene.
    Mol Cell Biol. 1990 Aug;10(8):3884-95 PMID: 2115115
  24. Role of NAD-linked glutamate dehydrogenase in nitrogen metabolism in Saccharomyces cerevisiae.
    J Bacteriol. 1990 Sep;172(9):4927-35 PMID: 1975578
  25. A mechanism for synergistic activation of a mammalian gene by GAL4 derivatives.
    Nature. 1990 May 24;345(6273):361-4 PMID: 2160609
  26. Release of two Saccharomyces cerevisiae cytochrome genes, COX6 and CYC1, from glucose repression requires the SNF1 and SSN6 gene products.
    Mol Cell Biol. 1990 Mar;10(3):1297-300 PMID: 2154683
  27. nit-2, the major positive-acting nitrogen regulatory gene of Neurospora crassa, encodes a sequence-specific DNA-binding protein.
    Proc Natl Acad Sci U S A. 1990 Jul;87(14):5331-5 PMID: 2142530
  28. How different eukaryotic transcriptional activators can cooperate promiscuously.
    Nature. 1990 May 24;345(6273):359-61 PMID: 2188137
  29. Regulation of glutamine synthetase from Saccharomyces cerevisiae by repression, inactivation and proteolysis.
    Eur J Biochem. 1982 Apr;123(3):611-6 PMID: 6122575
  30. An integrated and simplified approach to cloning into plasmids and single-stranded phages.
    Methods Enzymol. 1983;101:78-89 PMID: 6310343
  31. One-step gene disruption in yeast.
    Methods Enzymol. 1983;101:202-11 PMID: 6310324
  32. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast.
    Methods Enzymol. 1983;101:181-91 PMID: 6310321
  33. Distinctly regulated tandem upstream activation sites mediate catabolite repression of the CYC1 gene of S. cerevisiae.
    Cell. 1984 Feb;36(2):503-11 PMID: 6319028
  34. Heme regulates transcription of the CYC1 gene of S. cerevisiae via an upstream activation site.
    Cell. 1983 Apr;32(4):1279-86 PMID: 6301690
  35. A rapid boiling method for the preparation of bacterial plasmids.
    Anal Biochem. 1981 Jun;114(1):193-7 PMID: 6269464
  36. Transcription maps of adenovirus.
    Methods Enzymol. 1980;65(1):750-68 PMID: 6246372
  37. Transformation of intact yeast cells treated with alkali cations.
    J Bacteriol. 1983 Jan;153(1):163-8 PMID: 6336730
  38. Ammonia regulation of amino acid permeases in Saccharomyces cerevisiae.
    Mol Cell Biol. 1983 Apr;3(4):672-83 PMID: 6343842
  39. Isolation and characterization of mutants that produce the allantoin-degrading enzymes constitutively in Saccharomyces cerevisiae.
    Mol Cell Biol. 1982 Sep;2(9):1088-95 PMID: 6757722
  40. Mutants of yeast defective in sucrose utilization.
    Genetics. 1981 May;98(1):25-40 PMID: 7040163
  41. Of moulds and men, or two fingers are not better than one.
    Trends Genet. 1990 Oct;6(10):311-3 PMID: 2149222
  42. Nitrogen catabolite repression in yeasts and filamentous fungi.
    Adv Microb Physiol. 1985;26:1-88 PMID: 2869649
  43. The GLN1 locus of Saccharomyces cerevisiae encodes glutamine synthetase.
    Genetics. 1985 Oct;111(2):243-58 PMID: 2865193
  44. Mutational analysis of upstream activation sequence 2 of the CYC1 gene of Saccharomyces cerevisiae: a HAP2-HAP3-responsive site.
    Mol Cell Biol. 1988 Feb;8(2):647-54 PMID: 2832731
  45. Three regulatory systems control expression of glutamine synthetase in Saccharomyces cerevisiae at the level of transcription.
    Mol Gen Genet. 1989 Jun;217(2-3):370-7 PMID: 2570348
  46. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase.
    Cell. 1982 Jan;28(1):145-54 PMID: 7039847
  47. Nitrogen catabolite repression of asparaginase II in Saccharomyces cerevisiae.
    J Bacteriol. 1980 Jul;143(1):422-6 PMID: 6995441
  48. Regulation of compartmentation of amino acid pools in Saccharomyces cerevisiae and its effects on metabolic control.
    Eur J Biochem. 1980 Jul;108(2):439-47 PMID: 6997042
  49. Glutamine and ammonia in nitrogen catabolite repression of Saccharomyces cerevisiae.
    Biochem Biophys Res Commun. 1977 Mar 21;75(2):233-9 PMID: 322661
  50. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  51. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids.
    Cell. 1977 Nov;12(3):721-32 PMID: 922889
  52. Ammonia assimilation in Saccharomyces cerevisiae as mediated by the two glutamate dehydrogenases. Evidence for the gdhA locus being a structural gene for the NADP-dependent glutamate dehydrogenase.
    Mol Gen Genet. 1974;128(1):73-85 PMID: 4150855
  53. The amidotransferases.
    Adv Enzymol Relat Areas Mol Biol. 1973;39:91-183 PMID: 4355768
  54. Induction and repression of arginase and ornithine transaminase in baker's yeast.
    Antonie Van Leeuwenhoek. 1970;36(1):1-19 PMID: 4912187
  55. Yeast mutants pleiotropically impaired in the regulation of the two glutamate dehydrogenases.
    Biochem Biophys Res Commun. 1973 Jul 17;53(2):367-72 PMID: 4146147
  56. Glutamate synthase: properties of the reduced nicotinamide adenine dinucleotide-dependent enzyme from Saccharomyces cerevisiae.
    J Bacteriol. 1974 Apr;118(1):89-95 PMID: 4362465
  57. Nitrogen repression of the allantoin degradative enzymes in Saccharomyces cerevisiae.
    J Bacteriol. 1974 Jun;118(3):821-9 PMID: 4598006
  58. The DAL7 promoter consists of multiple elements that cooperatively mediate regulation of the gene's expression.
    Mol Cell Biol. 1989 Aug;9(8):3231-43 PMID: 2552287
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.12.6229-6247.1991
Published
1991-12-00
Pages
6229-47
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC361811
Subset
IM
Grants
NIGMS NIH HHS · GM-07446 · United States
Databases
GENBANK
M64559, M64560, M64561, M64562, M64563, M64564, M64565, M64566, S66436, X59842
External Links
PubMed source DOI: 10.1128/mcb.11.12.6229-6247.1991 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2