Summary
Mutants of Escherichia coli K12 requiring glutamine as a nitrogen source were isolated, and characterized as lacking glutamine synthetase activity. Temperature sensitive revertants of one of the mutants had a heat labile glutamine synthetase, while temperature insensitive revertants had a glutamine synthetase which was thermostable in vitro, indicating that the mutation was in the structural gene for the enzyme. All of the mutations mapped in the same region of the chromosome suggesting that they might all be in the same gene. The glutamine synthetase gene (gln) was located on the E. coli chromosome by conjugation and P1-mediated transduction at minute 77. The gln gene cotransduced with the genes for oleate degradation (old), and the genes for L-rhamnose utilization (rha). The most probable gene order is old-gln-rha.
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Brice, C. B., Kornberg, A. L.: Location of a gene specifying phosphopyruvate synthetase activity on the genome of Escherichia coli K12. Proc. roy. Soc. B 168, 281–292 (1967)
Brown, M., Segal, S. A., Stadtman, E. R.: Modulation of glutamine synthetase adenylylation and deadenylylation is mediated by metabolic transformation of the PII regulatory protein. Proc. nat. Acad. Sci. (Wash.) 68, 2949–2953 (1971)
Davis, B. D., Mingioli, E. S.: Mutants of Escherichia coli requiring methionine or vitamine B12. J. Bact. 60, 17–28 (1950)
Halpern, Y. S., Umbarger, H. E.: Conversion of ammonia to amino groups in Escherichia coli. J. Bact. 80, 285–288 (1960)
Kingdon, H. S., Hubbard, J. S., Stadtman, E. R.: Regulation of glutamine synthetase. XI. The nature and implications of a lag phase in the Escherichia coli glutamine synthetase reaction. Biochemistry (Wash.) 7, 2136–2142 (1968)
Kingdon, H. S., Noyes, C., Lahiri, A., Heinrikson, R. L.: Primary structure of Escherichia coli glutamine synthetase. III. Chemical evidence for subunit identy; amino acid sequence at the NH2 and COOH termini. J. biol. Chem. 247, 7923–7926 (1972)
Kingdon, H. S., Shapiro, B. M., Stadtman, E. R.: Regulation of glutamine synthetase. VIII. ATP: glutamine synthetase adenylyltransferase, an enzyme that catalyzes alterations in the regulatory properties of glutamine synthetase. Proc. nat. Acad. Sci. (Wash.) 58, 1703–1710 (1967)
Kingdon, H. S., Stadtman, E. R.: Two Escherichia coli glutamine synthetases with different sensitivities to feedback effectors. Biochem. biophys. Res. Commun. 27, 470–473 (1967)
Lennox, E. S.: Transduction of genetic characters of the host by bacteriophage P1. Virology 1, 190–206 (1955)
Leo, A. B. de, Magasanik, B.: Identification of the structural gene for glutamine synthetase in Klebsiella aerogenes. J. Bact. 121, 313–319 (1975)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the folin phenol reagent. J. biol. Chem. 193, 265–275 (1951)
Overath, P., Pauli, G., Schairer, H. U.: Fatty acid degradation in Escherichia coli. An inducible acyl-CoA synthetase, the mapping of old-mutations, and the isolation of regulatory mutants. Europ. J. Biochem. 7, 559–574 (1969)
Prival, M. J., Brenchley, J., Magasanik, B.: Glutamine synthetase and the regulation of histidine formation in Klebsiella aerogenes. J. biol. Chem. 248, 4334–4344 (1973)
Shapiro, B. M.: The glutamine synthetase deadenylylating enzyme system from Escherichia coli. Resolution into two components, specific nucleotide stimulation, and cofactor requirements. Biochemistry (Wash.) 8, 659–670 (1969)
Shapiro, B. M., Ginsburg, A.: Effects of specific divalent cations on some physical and chemical properties of glutamine synthetase from Escherichia coli. Taut and relaxed enzyme forms. Biochemistry (Wash.) 7, 2153–2167 (1968)
Shapiro, B. M., Kingdon, H. S., Stadtman, E. R.: Regulation of glutamine synthetase. VII. Adenylylglutamine synthetase: A new form of the enzyme with altered regulatory and kinetic properties. Proc. nat. Acad. Zci. (Wash.) 58, 642–649 (1967)
Shapiro, B. M., Stadtman, E. R.: Glutamine synthetase deadenylylating enzyme. Biochem. biophys. Res. Commun. 30, 32–37 (1968)
Stadtman, E. R., Ginsburg, A., Ciardi, J. E., Yeh, J., Hennig, S. B., Shapiro, B. M.: Multiple molecular forms of glutamine synthetase produced by enzyme catalyzed adenylylation and deadenylylation reactions. Advanc. Enz. Regul. 8, 99–118 (1969)
Sreicher, S. L., Bender, R. A., and Magasanik, B.: Genetic control of glutamine synthetase in Klebsiella aerogenes. J. Bact. 121, 320–331 (1975)
Swanstrom, M., Adams, M. H.: Agar-layer method for production of high titer phage stocks. Proc. Soc. exp. Biol. (N.Y.) 78, 372–375 (1951)
Taylor, A. L., Trotter, C. D.: Linkage map of Escherichia coli strain K12. Bact. Rev. 36, 504–524 (1972)
Woolfolk, C. A., Shapiro, B. M., Stadtman, E. R.: Regulation of glutamine synthetase. I. Purification and properties of glutamine synthetase from Escherichia coli. Arch. Biochem. Biophys. 116, 177–192 (1966)
Woolfolk, C. A., Stadtman, E. R.: Cummulative feedback inhibition in the multiple end-product regulation of glutamine synthetase activity in Escherichia coli. Biochem. biophys. Res. Commun. 17, 313–319 (1964)
Wu, C., Yuan, L. H.: Regulation of synthesis of glutamine synthetase in Escherichia coli. J. gen. Microbiol. 51, 57–65(1968)
Wulff, K., Mecke, D., Holzer, A.: Mechanism of enzymatic inactivation of glutamine synthetase from Escherichia coli. Biochem. biophys. Res. Commun. 28, 740–745 (1967)
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Communicated by W. Maas
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Mayer, E.P., Smith, O.H., Fredricks, W.W. et al. The isolation and characterization of glutamine-requiring strains of Escherichia coli K12. Molec. gen. Genet. 137, 131–142 (1975). https://doi.org/10.1007/BF00341679
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DOI: https://doi.org/10.1007/BF00341679