Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Sep;131(3):981–987. doi: 10.1128/jb.131.3.981-987.1977

Alteration of the Bacillus subtilis glutamine synthetase results in overproduction of the enzyme.

D R Dean, J A Hoch, A I Aronson
PMCID: PMC235556  PMID: 19424

Abstract

A mutational leading to glutamine auxotrophy was located near a 5-fluorouracil resistance marker in the citB-thyA region of the Bacillus subtilis chromosome. This mutation resulted in a glutamine synthetase with altered kinetic and feedback properties. The specific activity of manganese-stimulated glutamine synthetase activity in crude extracts was 18-fold higher, and the magnesium-stimulated activity was about 30% that of the wild type. Quantitation of the enzyme by precipitation with antibody prepared against pure enzyme confirmed the presence of high enzyme levels in the mutant. This mutation is very closely linked (recombination index of 0.03) to another glutamine auxotroph containing enzyme with altered electrophoretic and heat sensitivity properties. Mutations in the structural gene for glutamine synthetase may result not only in altered catalytic and regulatory properties but also in altered production of the enzyme.

Full text

PDF
985

Images in this article

985Image
on p.985

Selected References

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

  1. Anagnostopoulos C., Spizizen J. REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961 May;81(5):741–746. doi: 10.1128/jb.81.5.741-746.1961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BARAT M., ANAGNOSTOPOULOS C., SCHNEIDER A. M. LINKAGE RELATIONSHIPS OF GENES CONTROLLING ISOLEUCINE, VALINE, AND LEUCINE BIOSYNTHESIS IN BACILLUS SUBTILIS. J Bacteriol. 1965 Aug;90:357–369. doi: 10.1128/jb.90.2.357-369.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carlton B. C. Fine-structure mapping by transformation in the tryptophan region of Bacillus subtilis. J Bacteriol. 1966 May;91(5):1795–1803. doi: 10.1128/jb.91.5.1795-1803.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Deuel T. F. Bacillus subtilis glutamine synthetase. Specific catalytic changes associated with limited sulfhydryl modification. J Biol Chem. 1971 Feb 10;246(3):599–605. [PubMed] [Google Scholar]
  5. Deuel T. F., Ginsburg A., Yeh J., Shelton E., Stadtman E. R. Bacillus subtilis glutamine synthetase. Purification and physical characterization. J Biol Chem. 1970 Oct 25;245(20):5195–5205. [PubMed] [Google Scholar]
  6. Deuel T. F., Stadtman E. R. Some kinetic properties of Bacillus subtilis glutamine synthetase. J Biol Chem. 1970 Oct 25;245(20):5206–5213. [PubMed] [Google Scholar]
  7. Elmerich C. Le cycle du glutamate, point de départ du métabolisme de l'azote, chez Bacillus megaterium. Eur J Biochem. 1972 May 23;27(2):216–224. doi: 10.1111/j.1432-1033.1972.tb01829.x. [DOI] [PubMed] [Google Scholar]
  8. FREESE E., PARK S. W., CASHEL M. THE DEVELOPMENTAL SIGNIFICANCE OF ALANINE DEHYDROGENASE IN BACILLUS SUBTILIS. Proc Natl Acad Sci U S A. 1964 Jun;51:1164–1172. doi: 10.1073/pnas.51.6.1164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Kingdon H. S., Shapiro B. M., Stadtman E. R. Regulation of glutamine synthetase. 8. ATP: glutamine synthetase adenylyltransferase, an enzyme that catalyzes alterations in the regulatory properties of glutamine synthetase. Proc Natl Acad Sci U S A. 1967 Oct;58(4):1703–1710. doi: 10.1073/pnas.58.4.1703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Reysset G., Aubert J. P. Relationship between sporulation and mutations impairing glutamine synthetase in Bacillus megaterium. Biochem Biophys Res Commun. 1975 Aug 18;65(4):1237–1241. doi: 10.1016/s0006-291x(75)80362-7. [DOI] [PubMed] [Google Scholar]
  15. Schulp J. A., Stouthamer A. H. The influence of oxygen, glucose and nitrate upon the formation of nitrate reductase and the respiratory system in Bacillus licheniformis. J Gen Microbiol. 1970 Dec;64(2):195–203. doi: 10.1099/00221287-64-2-195. [DOI] [PubMed] [Google Scholar]
  16. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]

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

RESOURCES