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. 1980 Feb;141(2):671–679. doi: 10.1128/jb.141.2.671-679.1980

Isolation of Klebsiella aerogenes mutants cis-dominant for glutamine synthetase expression.

D M Rothstein, B Magasanik
PMCID: PMC293674  PMID: 6102550

Abstract

We have isolated three strains of Klebsiella aerogenes that failed to show repression of glutamine synthetase even when grown under the most repressing conditions for the wild-type strain. These mutant strains were selected as glutamine-independent derivatives of a strain that is merodiploid for the glnA region and contains a mutated glnF allele. The mutation responsible for the Gln+ phenotype in each strain was tightly linked to glnA, the structural gene for glutamine synthetase, and was dominant to the wild-type allele. These mutations are probably lesions in the control region of the glnA gene, since each mutation was cis-dominant for constitutive expression of the enzyme in hybrid merodiploid strains. Strains harboring this class of mutations were unable to produce a high level of glutamine synthetase unless they also contained an intact glnF gene, and unless cells were grown in derepressing medium. This study supports the idea that the glnA gene is regulated both positively and negatively, and that the deoxyribonucleic acid sites critical for positive control and negative control are functionally distinct.

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Selected References

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  1. 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]
  2. Bender R. A., Magasanik B. Autogenous regulation of the synthesis of glutamine synthetase in Klebsiella aerogenes. J Bacteriol. 1977 Oct;132(1):106–112. doi: 10.1128/jb.132.1.106-112.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Bender R. A., Streicher S. L. Glutamine synthetase regulation, adenylylation state, and strain specificity analyzed by polyacrylamide gel electrophoresis. J Bacteriol. 1979 Feb;137(2):1000–1007. doi: 10.1128/jb.137.2.1000-1007.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bloom F. R., Streicher S. L., Tyler B. Regulation of enzyme synthesis by the glutamine synthetase of Salmonella typhimurium: a factor in addition to glutamine synthetase is required for activation of enzyme formation. J Bacteriol. 1977 Jun;130(3):983–990. doi: 10.1128/jb.130.3.983-990.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Deleo A. B., Magasanik B. Identification of the structural gene for glutamine synthetase in Klebsiella aerogenes. J Bacteriol. 1975 Jan;121(1):313–319. doi: 10.1128/jb.121.1.313-319.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. 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]
  10. Garcia E., Bancroft S., Rhee S. G., Kustu S. The product of a newly identified gene, gInF, is required for synthesis of glutamine synthetase in Salmonella. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1662–1666. doi: 10.1073/pnas.74.4.1662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Janssen K. A., Magasanik B. Glutamine synthetase of Klebsiella aerogenes: genetic and physiological properties of mutants in the adenylylation system. J Bacteriol. 1977 Feb;129(2):993–1000. doi: 10.1128/jb.129.2.993-1000.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Meers J. L., Tempest D. W., Brown C. M. 'Glutamine(amide):2-oxoglutarate amino transferase oxido-reductase (NADP); an enzyme involved in the synthesis of glutamate by some bacteria. J Gen Microbiol. 1970 Dec;64(2):187–194. doi: 10.1099/00221287-64-2-187. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. 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]
  20. Shapiro B. M., Stadtman E. R. Regulation of glutamine synthetase. IX. Reactivity of the sulfhydryl groups of the enzyme from Escherichia coli. J Biol Chem. 1967 Nov 10;242(21):5069–5079. [PubMed] [Google Scholar]
  21. Smith G. R., Halpern Y. S., Magasanik B. Genetic and metabolic control of enzymes responsible for histidine degradation in Salmonella typhimurium. 4-imidazolone-5-propionate amidohydrolase and N-formimino-L-glutamate formiminohydrolase. J Biol Chem. 1971 May 25;246(10):3320–3329. [PubMed] [Google Scholar]
  22. 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]
  23. Streicher S. L., Deleo A. B., Magasanik B. Regulation of enzyme formation in Klebsiella aerogenes by episomal glutamine synthetase of Escherichia coli. J Bacteriol. 1976 Jul;127(1):184–192. doi: 10.1128/jb.127.1.184-192.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Tempest D. W., Meers J. L., Brown C. M. Synthesis of glutamate in Aerobacter aerogenes by a hitherto unknown route. Biochem J. 1970 Apr;117(2):405–407. doi: 10.1042/bj1170405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tyler B., Deleo A. B., Magasanik B. Activation of transcription of hut DNA by glutamine synthetase. Proc Natl Acad Sci U S A. 1974 Jan;71(1):225–229. doi: 10.1073/pnas.71.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]

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