Summary
76 mutants have been isolated in which the function of the single carbamoylphosphate synthetase of Escherichia coli K 12 is affected. A wide variety of phenotypes have been observed among these mutants, the most typical ones being: requirement for arginine and uracil, arginineless behaviour, sensitivity towards arginine and sensitivity towards uracil. The mutations have been localized by reciprocal transduction and deletion mapping; all are clustered in the same locus, car. The study of carbamoylphosphate synthesizing activities of these mutants and the combination of car mutations in various in vivo as well as in vitro complementation tests lead to the conclusion that car contains two genes: carA, covering the left part of the locus and coding for the “glutamine subunit” of the enzyme; carB, to the right, governing the synthesis of the heavy subunit of the enzyme.
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Abd-El-Al, A., Ingraham, J. L.: Control of carbamylphosphate synthesis in Salmonella typhimurium. J. biol. Chem. 244, 4033–4038 (1969a)
Abd-El-Al, A., Ingraham, J. L.: Cold sensitivity and other phenotypes resulting from mutation in pyrA gene. J. biol. Chem. 244, 4039–4045 (1969b)
Abd-El-Al, A., Kessler, D. P., Ingraham, J. L.: Arginine-auxotrophic phenotype resulting from a mutation in the pyrA gene of Escherichia coli b/r. J. Bact. 97, 466–468 (1969)
Adelberg, E. A., Mandel, M., Chein Ching Chen, G.: Optimal conditions for mutagenesis by N-methyl-N′-nitro-N-nitrosoguanidine in Escherichia coli K12. Biochem. biophys. Res. Commun. 18, 788–795 (1965)
Anderson, P. M., Meister, A.: Evidence for an actvated form of carbon dioxide in the reaction catalyzed by Escherichia coli carbamylphosphate synthetase. Biochemistry 4, 2803–2809 (1965)
Anderson, P. M., Meister, A.: Control of carbamylphosphate synthetase by purine and pyrimidine nucleotides. Biochemistry 5, 3164–3169 (1966)
Anderson, P. M., Marvin, S. V.: Effect of allosteric effectors and adenosine triphosphate on the aggregation and rate of inhibition by N-ethylmaleimide of carbamylphosphate synthetase of Escherichia coli. Biochemistry 9, 171–178 (1970)
Bachman, B. J.: Pedigrees of some mutant strains of Escherichia coli K12. Bact. Rev. 36, 525–557 (1972)
Beckwith, J. R., Pardee, A. B., Austrian, R., Jacob, F.: Coordination of the synthesis of the enzymes in the pyrimidine pathway of E. coli. J. molec. Biol. 5, 618–634 (1962)
Charles, H. P., Roberts, G. A.: Carbon dioxide as a growth factor for mutants of Escherichia coli. J. gen. Microbiol. 51, 211–224 (1968)
Davis, R. H.: Channeling in Neurospora metabolism. In: Organizational biosynthesis (ed. J. H. Vogel, J. O. Lampen and V. Bryzon), p. 303–322. New York: Academic Press 1967
Eisenstark, A.: Linkage of arginine sensitive (ars) and uracil-arginine requiring (pyrA) loci of Salmonella typhimurium. Nature (Lond.) 213, 1263–1264 (1967)
Glansdorff, N.: Topography of cotransducible arginine mutations in Escherichia coli K12. Genetics 51, 167–179 (1965)
Gorini, L., Kalman, S. M.: Control by uracil of carbamylphosphate synthesis in Escherichia coli. Biochim. biophys. Acta (Amst.) 69, 355–360 (1963)
Gorini, L., Kaufman, H.: Selecting bacterial mutants by the penicillin method. Science 131, 604–605 (1960)
Hartman, P. E., Hartman, Z., Serman, D.: Complementation mapping by abortive transduction of histidine requiring mutants. J. gen. Microbiol. 22, 354–368 (1960)
Hopwood, D. A.: Genetic analysis and genome structure in Streptomyces coelicolor. Bact. Rev. 31, 373–403 (1969)
Issaly, I. M., Issaly, A. S., Reissig, J. L.: Carbamylphosphate biosynthesis in Bacillus subtilis. Biochim. biophys. Acta (Amst.) 198, 482–494 (1970)
Jacob, F.: Transduction of lysogeny in Escherichia coli. Virology 1, 207–220 (1955)
Kalman, S. M., Duffield, P. H., Brozozowski, I.: Purification and properties of a bacterial carbamylphosphate synthetase. J. biol. Chem. 241, 1871–1877 (1966)
Khedouri, E., Anderson, P. M., Meister, A.: Selective inactivation of the glutamine binding site of Escherichia coli carbamylphosphate synthetase by 2-amino-4-oxo-5-chloropentanoic acid. Biochemistry 5, 3552–3556 (1966)
Loutit, J. S.: Studies on nutritionally deficient strains of Pseudomonas aeruginosa. 1. The production by X-rays and the isolation of nutritionally deficient strains. Aust. J. exp. Biol. med. Sci. 30, 287–294 (1952)
Low, B.: Formation of merodiploids in mating with a class of rec- recipient strain of Escherichia coli K12. Proc. nat. Acad. Sci. (Wash.) 60, 160–167 (1968)
Martin, R. G.: Frameshift mutants in the histidine operon of Salmonella typhimurium. J. molec. Biol. 26, 311–328 (1967)
Mergeay, M.: Physiologie et génétique d'un branchement métabolique: la biosynthèse du carbamylphosphate chez Escherichia coli. Thèse de doctorat, Université Libre de Bruxelles (1969)
Novick, P. R., Maas, W. K.: Control by endogenously synthesized arginine of the formation of ornithine transcarbamylase in Escherichia coli. J. Bact. 81, 236–240 (1961)
Oeschger, N. S., Hartman, P. E.: ICR-induced frameshift mutations in the histidine operon of Salmonella. J. Bact. 101, 490–504 (1970)
Piérard, A.: Control of the activity of Escherichia coli carbamoylphosphate synthetase by antagonistic allosteric effectors. Science 154, 1572–1573 (1966)
Piérard, A., Glansdorff, N., Mergeay, M., Wiame, J.-M.: Control of the biosynthesis of carbamoylphosphate in Escherichia coli. J. molec. Biol. 14, 23–26 (1965)
Piérard, A., Glansdorff, N., Yashphe, J.: Mutations affecting uridine monophosphate pyrophosphorylase or the argR gene in Escherichia coli. Effects on carbamoylphosphate and pyrimidine biosynthesis and on uracil uptake. Molec. gen. Genet. 118, 235–245 (1972)
Piérard, A., Grenson, M., Glansdorff, N., Wiame, J.-M.: A comparison of the organization of carbamoylphosphate synthesis in Saccharomyces cerevisiae and Escherichia coli, based on genetical and biochemical evidences. In: The enzymes of glutamine metabolism (ed. S. Prusiner and E. R. Stadtman), p. 483–503. New York: Academic Press 1973
Piérard, A., Wiame, J.-M.: Regulation and mutation affecting a glutamine dependent formation of carbamylphosphate in Escherichia coli. Biochem. biophys. Res. Commun. 15, 76–81 (1964)
Pinkus, M. L., Meister, A.: Identification of a reactive cystein residue at the glutamine binding site of carbamylphosphate synthetase. J. biol. Chem. 247, 6119–6127 (1972)
Prozesky, O. M., Coetzee, J. N.: Linked transduction in Proteus mirabilis. Nature (Lond.) 209, 1262 (1966)
Roepke, R. R., Mercer, F. E.: Lethal and sublethal effects of X rays on Escherichia coli as related to the yield of biochemical mutant. J. Bact. 54, 731–743 (1947)
Syvanen, J. M., Roth, J. R.: Structural genes for catalytic and regulatory subunits of aspartate transcarbamylase. J. molec. Biol. 76, 363–368 (1973)
Taylor, A. L., Trotter, C. D.: Linkage map of Escherichia coli strain K-12. Bact. Rev. 36, 504–524 (1972)
Trotta, P. P., Burt, M. E., Haschenmeyer, R. H., Meister, A.: Reversible dissociation of carbamylphosphate synthetase into a regulated synthesis subunit and a subunit required for glutamine utilization. Proc. nat. Acad. Sci. (Wash.) 68, 2599–2603 (1971)
Van Montagu, M., Leurs, C., Brachet, P., Thomas, R.: A set of amber mutants of bacteriophages lambda and MS2 suitable for the identification of suppressors. Mutation Res. 4, 698–700 (1967)
Vyas, S., Maas, W. K.: Feedback inhibition of acetylglutamate synthetase by arginine in Escherichia coli. Arch. Biochem. Biophys. 100, 542–546 (1963)
Whitfield, H. J., Martin, R. G., Ames, B. N.: Classification of aminotransferase (C genes) mutants in the histidine operon. J. molec. Biol. 21, 335–355 (1966)
Yan, Y., Demerec, M.: Genetic analysis of pyrimidine mutants of Salmonella typhimurium. Genetics 52, 643–651 (1965)
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Mergeay, M., Gigot, D., Beckmann, J. et al. Physiology and genetics of carbamoylphosphate synthesis in Escherichia coli K12. Molec. gen. Genet. 133, 299–316 (1974). https://doi.org/10.1007/BF00332706
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DOI: https://doi.org/10.1007/BF00332706