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. 1995 Jan;177(1):94–102. doi: 10.1128/jb.177.1.94-102.1995

Transcriptional and posttranslational regulation of the general amino acid permease of Saccharomyces cerevisiae.

M Stanbrough 1, B Magasanik 1
PMCID: PMC176561  PMID: 7798155

Abstract

The cellular level and activity of the general amino acid permease, the product of the GAP1 gene of Saccharomyces cerevisiae, are regulated at the level of transcription by two systems, the products of URE2/GLN3 and NIL1 in response to the nitrogen sources of the growth medium and inactivation in response to the presence of glutamine or glutamate. Active permease is phosphorylated. The addition of glutamine causes rapid dephosphorylation and inactivation of the permease with the same kinetics, which is followed by slower disappearance of the protein. These results suggest that inactivation of the permease results from its dephosphorylation.

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Selected References

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  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandriss M. C., Magasanik B. Proline: an essential intermediate in arginine degradation in Saccharomyces cerevisiae. J Bacteriol. 1980 Sep;143(3):1403–1410. doi: 10.1128/jb.143.3.1403-1410.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carlson M., Botstein D. Two differentially regulated mRNAs with different 5' ends encode secreted with intracellular forms of yeast invertase. Cell. 1982 Jan;28(1):145–154. doi: 10.1016/0092-8674(82)90384-1. [DOI] [PubMed] [Google Scholar]
  4. Clark-Adams C. D., Winston F. The SPT6 gene is essential for growth and is required for delta-mediated transcription in Saccharomyces cerevisiae. Mol Cell Biol. 1987 Feb;7(2):679–686. doi: 10.1128/mcb.7.2.679-686.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Coschigano P. W., Magasanik B. 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):822–832. doi: 10.1128/mcb.11.2.822-832.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Courchesne W. E., Magasanik B. Ammonia regulation of amino acid permeases in Saccharomyces cerevisiae. Mol Cell Biol. 1983 Apr;3(4):672–683. doi: 10.1128/mcb.3.4.672-683.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Courchesne W. E., Magasanik B. Regulation of nitrogen assimilation in Saccharomyces cerevisiae: roles of the URE2 and GLN3 genes. J Bacteriol. 1988 Feb;170(2):708–713. doi: 10.1128/jb.170.2.708-713.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Forsburg S. L., Guarente L. 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):647–654. doi: 10.1128/mcb.8.2.647-654.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fu Y. H., Marzluf G. A. Characterization of nit-2, the major nitrogen regulatory gene of Neurospora crassa. Mol Cell Biol. 1987 May;7(5):1691–1696. doi: 10.1128/mcb.7.5.1691-1696.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fu Y. H., Marzluf G. A. 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):5331–5335. doi: 10.1073/pnas.87.14.5331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Grenson M., Hou C. Ammonia inhibition of the general amino acid permease and its suppression in NADPH-specific glutamate dehydrogenaseless mutants of saccharomyces cerevisiae. Biochem Biophys Res Commun. 1972 Aug 21;48(4):749–756. doi: 10.1016/0006-291x(72)90670-5. [DOI] [PubMed] [Google Scholar]
  12. Grenson M., Hou C., Crabeel M. Multiplicity of the amino acid permeases in Saccharomyces cerevisiae. IV. Evidence for a general amino acid permease. J Bacteriol. 1970 Sep;103(3):770–777. doi: 10.1128/jb.103.3.770-777.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Grenson M. Inactivation-reactivation process and repression of permease formation regulate several ammonia-sensitive permeases in the yeast Saccharomyces cerevisiae. Eur J Biochem. 1983 Jun 1;133(1):135–139. doi: 10.1111/j.1432-1033.1983.tb07438.x. [DOI] [PubMed] [Google Scholar]
  14. Grenson M. Study of the positive control of the general amino-acid permease and other ammonia-sensitive uptake systems by the product of the NPR1 gene in the yeast Saccharomyces cerevisiae. Eur J Biochem. 1983 Jun 1;133(1):141–144. doi: 10.1111/j.1432-1033.1983.tb07439.x. [DOI] [PubMed] [Google Scholar]
  15. Haldi M., Guarente L. N-terminal deletions of a mitochondrial signal sequence in yeast. Targeting information of delta-aminolevulinate synthase is encoded in non-overlapping regions. J Biol Chem. 1989 Oct 15;264(29):17107–17112. [PubMed] [Google Scholar]
  16. Huisman O., Raymond W., Froehlich K. U., Errada P., Kleckner N., Botstein D., Hoyt M. A. A Tn10-lacZ-kanR-URA3 gene fusion transposon for insertion mutagenesis and fusion analysis of yeast and bacterial genes. Genetics. 1987 Jun;116(2):191–199. doi: 10.1093/genetics/116.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Jauniaux J. C., Grenson M. GAP1, the general amino acid permease gene of Saccharomyces cerevisiae. Nucleotide sequence, protein similarity with the other bakers yeast amino acid permeases, and nitrogen catabolite repression. Eur J Biochem. 1990 May 31;190(1):39–44. doi: 10.1111/j.1432-1033.1990.tb15542.x. [DOI] [PubMed] [Google Scholar]
  19. Kolodziej P. A., Young R. A. Epitope tagging and protein surveillance. Methods Enzymol. 1991;194:508–519. doi: 10.1016/0076-6879(91)94038-e. [DOI] [PubMed] [Google Scholar]
  20. Kudla B., Caddick M. X., Langdon T., Martinez-Rossi N. M., Bennett C. F., Sibley S., Davies R. W., Arst H. N., Jr The regulatory gene areA mediating nitrogen metabolite repression in Aspergillus nidulans. Mutations affecting specificity of gene activation alter a loop residue of a putative zinc finger. EMBO J. 1990 May;9(5):1355–1364. doi: 10.1002/j.1460-2075.1990.tb08250.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ljungdahl P. O., Gimeno C. J., Styles C. A., Fink G. R. SHR3: a novel component of the secretory pathway specifically required for localization of amino acid permeases in yeast. Cell. 1992 Oct 30;71(3):463–478. doi: 10.1016/0092-8674(92)90515-e. [DOI] [PubMed] [Google Scholar]
  22. Miller S. M., Magasanik B. Role of NAD-linked glutamate dehydrogenase in nitrogen metabolism in Saccharomyces cerevisiae. J Bacteriol. 1990 Sep;172(9):4927–4935. doi: 10.1128/jb.172.9.4927-4935.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Miller S. M., Magasanik B. Role of the complex upstream region of the GDH2 gene in nitrogen regulation of the NAD-linked glutamate dehydrogenase in Saccharomyces cerevisiae. Mol Cell Biol. 1991 Dec;11(12):6229–6247. doi: 10.1128/mcb.11.12.6229-6247.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Minehart P. L., Magasanik B. Sequence of the GLN1 gene of Saccharomyces cerevisiae: role of the upstream region in regulation of glutamine synthetase expression. J Bacteriol. 1992 Mar;174(6):1828–1836. doi: 10.1128/jb.174.6.1828-1836.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mitchell A. P., Magasanik B. Purification and properties of glutamine synthetase from Saccharomyces cerevisiae. J Biol Chem. 1983 Jan 10;258(1):119–124. [PubMed] [Google Scholar]
  26. Plumb M., Frampton J., Wainwright H., Walker M., Macleod K., Goodwin G., Harrison P. GATAAG; a cis-control region binding an erythroid-specific nuclear factor with a role in globin and non-globin gene expression. Nucleic Acids Res. 1989 Jan 11;17(1):73–92. doi: 10.1093/nar/17.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rytka J. Positive selection of general amino acid permease mutants in Saccharomyces cerevisiae. J Bacteriol. 1975 Feb;121(2):562–570. doi: 10.1128/jb.121.2.562-570.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sadowski I., Niedbala D., Wood K., Ptashne M. GAL4 is phosphorylated as a consequence of transcriptional activation. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10510–10514. doi: 10.1073/pnas.88.23.10510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Silve S., Volland C., Garnier C., Jund R., Chevallier M. R., Haguenauer-Tsapis R. Membrane insertion of uracil permease, a polytopic yeast plasma membrane protein. Mol Cell Biol. 1991 Feb;11(2):1114–1124. doi: 10.1128/mcb.11.2.1114-1124.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Strathern J. N., Higgins D. R. Recovery of plasmids from yeast into Escherichia coli: shuttle vectors. Methods Enzymol. 1991;194:319–329. doi: 10.1016/0076-6879(91)94024-7. [DOI] [PubMed] [Google Scholar]
  32. Sumrada R. A., Cooper T. G. 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. doi: 10.1073/pnas.84.12.3997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Tsai S. F., Martin D. I., Zon L. I., D'Andrea A. D., Wong G. G., Orkin S. H. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells. Nature. 1989 Jun 8;339(6224):446–451. doi: 10.1038/339446a0. [DOI] [PubMed] [Google Scholar]
  34. Vandenbol M., Jauniaux J. C., Grenson M. The Saccharomyces cerevisiae NPR1 gene required for the activity of ammonia-sensitive amino acid permeases encodes a protein kinase homologue. Mol Gen Genet. 1990 Jul;222(2-3):393–399. doi: 10.1007/BF00633845. [DOI] [PubMed] [Google Scholar]
  35. Vandenbol M., Jauniaux J. C., Vissers S., Grenson M. Isolation of the NPR1 gene responsible for the reactivation of ammonia-sensitive amino-acid permeases in Saccharomyces cerevisiae. RNA analysis and gene dosage effects. Eur J Biochem. 1987 May 4;164(3):607–612. doi: 10.1111/j.1432-1033.1987.tb11170.x. [DOI] [PubMed] [Google Scholar]
  36. Wiame J. M., Grenson M., Arst H. N., Jr Nitrogen catabolite repression in yeasts and filamentous fungi. Adv Microb Physiol. 1985;26:1–88. doi: 10.1016/s0065-2911(08)60394-x. [DOI] [PubMed] [Google Scholar]

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