Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1990 Dec;172(12):7249–7255. doi: 10.1128/jb.172.12.7249-7255.1990

Role of the nac gene product in the nitrogen regulation of some NTR-regulated operons of Klebsiella aerogenes.

A Macaluso 1, E A Best 1, R A Bender 1
PMCID: PMC210849  PMID: 1979323

Abstract

A positive, genetic selection against the activity of the nitrogen regulatory (NTR) system was used to isolate insertion mutations affecting nitrogen regulation in Klebsiella aerogenes. Two classes of mutation were obtained: those affecting the NTR system itself and leading to the loss of almost all nitrogen regulation, and those affecting the nac locus and leading to a loss of nitrogen regulation of a family of nitrogen-regulated enzymes. The set of these nac-dependent enzymes included histidase, glutamate dehydrogenase, glutamate synthase, proline oxidase, and urease. The enzymes shown to be nac independent included glutamine synthetase, asparaginase, tryptophan permease, nitrate reductase, the product of the nifLA operon, and perhaps nitrite reductase. The expression of the nac gene was itself highly nitrogen regulated, and this regulation was mediated by the NTR system. The loss of nitrogen regulation was found in each of the four insertion mutants studied, showing that loss of nitrogen regulation resulted from the absence of nac function rather than from an altered form of the nac gene product. Thus we propose two classes of nitrogen-regulated operons: in class I, the NTR system directly activates expression of the operon; in class II, the NTR system activates nac expression and the product(s) of the nac locus activates expression of the operon.

Full text

PDF
7249

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Austin S., Henderson N., Dixon R. Requirements for transcriptional activation in vitro of the nitrogen-regulated glnA and nifLA promoters from Klebsiella pneumoniae: dependence on activator concentration. Mol Microbiol. 1987 Jul;1(1):92–100. doi: 10.1111/j.1365-2958.1987.tb00532.x. [DOI] [PubMed] [Google Scholar]
  2. Baker T. A., Howe M. M., Gross C. A. Mu dX, a derivative of Mu d1 (lac Apr) which makes stable lacZ fusions at high temperature. J Bacteriol. 1983 Nov;156(2):970–974. doi: 10.1128/jb.156.2.970-974.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baldauf S. L., Cardani M. A., Bender R. A. Regulation of the galactose-inducible lac operon and the histidine utilization operons in pts mutants of Klebsiella aerogenes. J Bacteriol. 1988 Dec;170(12):5588–5593. doi: 10.1128/jb.170.12.5588-5593.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bender R. A., Eades L. J. A nonsense mutation in the structural gene for glutamine synthetase leading to loss of nitrogen regulation in Klebsiella aerogenes. Mol Gen Genet. 1982;187(3):414–418. doi: 10.1007/BF00332621. [DOI] [PubMed] [Google Scholar]
  5. Bender R. A., Janssen K. A., Resnick A. D., Blumenberg M., Foor F., Magasanik B. Biochemical parameters of glutamine synthetase from Klebsiella aerogenes. J Bacteriol. 1977 Feb;129(2):1001–1009. doi: 10.1128/jb.129.2.1001-1009.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bender R. A., Snyder P. M., Bueno R., Quinto M., Magasanik B. Nitrogen regulation system of Klebsiella aerogenes: the nac gene. J Bacteriol. 1983 Oct;156(1):444–446. doi: 10.1128/jb.156.1.444-446.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bremer E., Silhavy T. J., Weinstock G. M. Transposable lambda placMu bacteriophages for creating lacZ operon fusions and kanamycin resistance insertions in Escherichia coli. J Bacteriol. 1985 Jun;162(3):1092–1099. doi: 10.1128/jb.162.3.1092-1099.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Brenchley J. E., Prival M. J., Magasanik B. Regulation of the synthesis of enzymes responsible for glutamate formation in Klebsiella aerogenes. J Biol Chem. 1973 Sep 10;248(17):6122–6128. [PubMed] [Google Scholar]
  9. Egner C., Berg D. E. Excision of transposon Tn5 is dependent on the inverted repeats but not on the transposase function of Tn5. Proc Natl Acad Sci U S A. 1981 Jan;78(1):459–463. doi: 10.1073/pnas.78.1.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Friedrich B., Magasanik B. Urease of Klebsiella aerogenes: control of its synthesis by glutamine synthetase. J Bacteriol. 1977 Aug;131(2):446–452. doi: 10.1128/jb.131.2.446-452.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gaillardin C. M., Magasanik B. Involvement of the product of the glnF gene in the autogenous regulation of glutamine synthetase formation in Klebsiella aerogenes. J Bacteriol. 1978 Mar;133(3):1329–1338. doi: 10.1128/jb.133.3.1329-1338.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Goldberg R. B., Bender R. A., Streicher S. L. Direct selection for P1-sensitive mutants of enteric bacteria. J Bacteriol. 1974 Jun;118(3):810–814. doi: 10.1128/jb.118.3.810-814.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goldie H., Magasanik B. Effects of glnL and other regulatory loci on regulation of transcription of glnA-lacZ fusions in Klebsiella aerogenes. J Bacteriol. 1982 Apr;150(1):231–238. doi: 10.1128/jb.150.1.231-238.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hirschman J., Wong P. K., Sei K., Keener J., Kustu S. Products of nitrogen regulatory genes ntrA and ntrC of enteric bacteria activate glnA transcription in vitro: evidence that the ntrA product is a sigma factor. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7525–7529. doi: 10.1073/pnas.82.22.7525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hunt T. P., Magasanik B. Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8453–8457. doi: 10.1073/pnas.82.24.8453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Lester R. L., DeMoss J. A. Effects of molybdate and selenite on formate and nitrate metabolism in Escherichia coli. J Bacteriol. 1971 Mar;105(3):1006–1014. doi: 10.1128/jb.105.3.1006-1014.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Magasanik B. Genetic control of nitrogen assimilation in bacteria. Annu Rev Genet. 1982;16:135–168. doi: 10.1146/annurev.ge.16.120182.001031. [DOI] [PubMed] [Google Scholar]
  19. Magasanik B., Prival M. J., Brenchley J. E., Tyler B. M., DeLeo A. B., Streicher S. L., Bender R. A., Paris C. G. Glutamine synthetase as a regulator of enzyme synthesis. Curr Top Cell Regul. 1974;8(0):119–138. doi: 10.1016/b978-0-12-152808-9.50010-9. [DOI] [PubMed] [Google Scholar]
  20. Nieuwkoop A. J., Baldauf S. A., Hudspeth M. E., Bender R. A. Bidirectional promoter in the hut(P) region of the histidine utilization (hut) operons from Klebsiella aerogenes. J Bacteriol. 1988 May;170(5):2240–2246. doi: 10.1128/jb.170.5.2240-2246.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ninfa A. J., Magasanik B. Covalent modification of the glnG product, NRI, by the glnL product, NRII, regulates the transcription of the glnALG operon in Escherichia coli. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5909–5913. doi: 10.1073/pnas.83.16.5909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ninfa A. J., Reitzer L. J., Magasanik B. Initiation of transcription at the bacterial glnAp2 promoter by purified E. coli components is facilitated by enhancers. Cell. 1987 Sep 25;50(7):1039–1046. doi: 10.1016/0092-8674(87)90170-x. [DOI] [PubMed] [Google Scholar]
  23. O'Neill E. A., Kiely G. M., Bender R. A. Transposon Tn5 encodes streptomycin resistance in nonenteric bacteria. J Bacteriol. 1984 Jul;159(1):388–389. doi: 10.1128/jb.159.1.388-389.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ow D. W., Ausubel F. M. Regulation of nitrogen metabolism genes by nifA gene product in Klebsiella pneumoniae. Nature. 1983 Jan 27;301(5898):307–313. doi: 10.1038/301307a0. [DOI] [PubMed] [Google Scholar]
  25. Paris C. G., Magasanik B. Tryptophan metabolism in Klebsiella aerogenes: regulation of the utilization of aromatic amino acids as sources of nitrogen. J Bacteriol. 1981 Jan;145(1):257–265. doi: 10.1128/jb.145.1.257-265.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Popham D. L., Szeto D., Keener J., Kustu S. Function of a bacterial activator protein that binds to transcriptional enhancers. Science. 1989 Feb 3;243(4891):629–635. doi: 10.1126/science.2563595. [DOI] [PubMed] [Google Scholar]
  27. Prival M. J., Brenchley J. E., Magasanik B. Glutamine synthetase and the regulation of histidase formation in Klebsiella aerogenes. J Biol Chem. 1973 Jun 25;248(12):4334–4344. [PubMed] [Google Scholar]
  28. Prival M. J., Magasanik B. Resistance to catabolite repression of histidase and proline oxidase during nitrogen-limited growth of Klebsiella aerogenes. J Biol Chem. 1971 Oct 25;246(20):6288–6296. [PubMed] [Google Scholar]
  29. Quinto M., Bender R. A. Use of bacteriophage P1 as a vector for Tn5 insertion mutagenesis. Appl Environ Microbiol. 1984 Feb;47(2):436–438. doi: 10.1128/aem.47.2.436-438.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Resnick A. D., Magasanik B. L-Asparaginase of Klebsiella aerogenes. Activation of its synthesis by glutamine synthetase. J Biol Chem. 1976 May 10;251(9):2722–2728. [PubMed] [Google Scholar]
  31. de Vries G. E., Raymond C. K., Ludwig R. A. Extension of bacteriophage lambda host range: selection, cloning, and characterization of a constitutive lambda receptor gene. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6080–6084. doi: 10.1073/pnas.81.19.6080. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES