Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1993 Apr;175(7):2116–2124. doi: 10.1128/jb.175.7.2116-2124.1993

The product of the Klebsiella aerogenes nac (nitrogen assimilation control) gene is sufficient for activation of the hut operons and repression of the gdh operon.

A Schwacha 1, R A Bender 1
PMCID: PMC204320  PMID: 8458854

Abstract

In Klebsiella aerogenes, the formation of a large number of enzymes responds to the quality and quantity of the nitrogen source provided in the growth medium, and this regulation requires the action of the nitrogen regulatory (NTR) system in every case known. Nitrogen regulation of several operons requires not only the NTR system, but also NAC, the product of the nac gene, raising the question of whether the role of NAC is to activate operons directly or by modifying the specificity of the NTR system. We isolated an insertion of the transposon Tn5tac1 which puts nac gene expression under the control of the IPTG-inducible tac promoter rather than the nitrogen-responsive nac promoter. When IPTG was present, cells carrying the tac-nac fusion activated NAC-dependent operons and repressed NAC-repressible operons independent of the nitrogen supply and even in the absence of an active NTR system. Thus, NAC is sufficient to regulate operons like hut (encoding histidase) and gdh (encoding glutamate dehydrogenase), confirming the model that the NTR system activates nac expression and NAC activates hut and represses gdh. Activation of urease formation occurred at a lower level of NAC than that required for glutamate dehydrogenase repression, and activation of histidase formation required still more NAC.

Full text

PDF
2116

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. 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]
  3. Bender R. A., Friedrich B. Regulation of assimilatory nitrate reductase formation in Klebsiella aerogenes W70. J Bacteriol. 1990 Dec;172(12):7256–7259. doi: 10.1128/jb.172.12.7256-7259.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Bender R. A., Magasanik B. Regulatory mutations in the Klebsiella aerogenes structural gene for glutamine synthetase. J Bacteriol. 1977 Oct;132(1):100–105. doi: 10.1128/jb.132.1.100-105.1977. [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. Bender R. A. The role of the NAC protein in the nitrogen regulation of Klebsiella aerogenes. Mol Microbiol. 1991 Nov;5(11):2575–2580. doi: 10.1111/j.1365-2958.1991.tb01965.x. [DOI] [PubMed] [Google Scholar]
  8. Best E. A., Bender R. A. Cloning of the Klebsiella aerogenes nac gene, which encodes a factor required for nitrogen regulation of the histidine utilization (hut) operons in Salmonella typhimurium. J Bacteriol. 1990 Dec;172(12):7043–7048. doi: 10.1128/jb.172.12.7043-7048.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Chen L. M., Maloy S. Regulation of proline utilization in enteric bacteria: cloning and characterization of the Klebsiella put control region. J Bacteriol. 1991 Jan;173(2):783–790. doi: 10.1128/jb.173.2.783-790.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chow W. Y., Berg D. E. Tn5tac1, a derivative of transposon Tn5 that generates conditional mutations. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6468–6472. doi: 10.1073/pnas.85.17.6468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Foor F., Cedergren R. J., Streicher S. L., Rhee S. G., Magasanik B. Glutamine synthetase of Klebsiella aerogenes: properties of glnD mutants lacking uridylyltransferase. J Bacteriol. 1978 May;134(2):562–568. doi: 10.1128/jb.134.2.562-568.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Foor F., Janssen K. A., Magasanik B. Regulation of synthesis of glutamine synthetase by adenylylated glutamine synthetase. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4844–4848. doi: 10.1073/pnas.72.12.4844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Foor F., Reuveny Z., Magasanik B. Regulation of the synthesis of glutamine synthetase by the PII protein in Klebsiella aerogenes. Proc Natl Acad Sci U S A. 1980 May;77(5):2636–2640. doi: 10.1073/pnas.77.5.2636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. Holtel A., Merrick M. Identification of the Klebsiella pneumoniae glnB gene: nucleotide sequence of wild-type and mutant alleles. Mol Gen Genet. 1988 Dec;215(1):134–138. doi: 10.1007/BF00331314. [DOI] [PubMed] [Google Scholar]
  19. Kustu S., Burton D., Garcia E., McCarter L., McFarland N. Nitrogen control in Salmonella: regulation by the glnR and glnF gene products. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4576–4580. doi: 10.1073/pnas.76.9.4576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Lanzer M., Bujard H. Promoters largely determine the efficiency of repressor action. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8973–8977. doi: 10.1073/pnas.85.23.8973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Macaluso A., Best E. A., Bender R. A. Role of the nac gene product in the nitrogen regulation of some NTR-regulated operons of Klebsiella aerogenes. J Bacteriol. 1990 Dec;172(12):7249–7255. doi: 10.1128/jb.172.12.7249-7255.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. NEIDHARDT F. C., MAGASANIK B. Reversal of the glucose inhibition of histidase biosynthesis in Aerobacter aerogenes. J Bacteriol. 1957 Feb;73(2):253–259. doi: 10.1128/jb.73.2.253-259.1957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Nieuwkoop A. J., Bender R. A. RNA polymerase as a repressor of transcription in the hut(P) region of mutant Klebsiella aerogenes histidine utilization operons. J Bacteriol. 1988 Oct;170(10):4986–4990. doi: 10.1128/jb.170.10.4986-4990.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Pahel G., Tyler B. A new glnA-linked regulatory gene for glutamine synthetase in Escherichia coli. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4544–4548. doi: 10.1073/pnas.76.9.4544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. 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]
  30. Schwacha A., Bender R. A. The nac (nitrogen assimilation control) gene from Klebsiella aerogenes. J Bacteriol. 1993 Apr;175(7):2107–2115. doi: 10.1128/jb.175.7.2107-2115.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Simon R. High frequency mobilization of gram-negative bacterial replicons by the in vitro constructed Tn5-Mob transposon. Mol Gen Genet. 1984;196(3):413–420. doi: 10.1007/BF00436188. [DOI] [PubMed] [Google Scholar]
  32. Stock J. B., Ninfa A. J., Stock A. M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. doi: 10.1128/mr.53.4.450-490.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Streicher S. L., Bender R. A., Magasanik B. Genetic control of glutamine synthetase in Klebiella aerogenes. J Bacteriol. 1975 Jan;121(1):320–331. doi: 10.1128/jb.121.1.320-331.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Weiss V., Claverie-Martin F., Magasanik B. Phosphorylation of nitrogen regulator I of Escherichia coli induces strong cooperative binding to DNA essential for activation of transcription. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5088–5092. doi: 10.1073/pnas.89.11.5088. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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