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Molecular and General Genetics MGG

, Volume 194, Issue 1–2, pp 114–123 | Cite as

cis-dominant, glutamine synthetase constitutive mutations of Escherichia coli indepedent of activation by the glnG and glnF products

  • Aurora V. Osorio
  • Luis Servín-González
  • Mario Rocha
  • Alejandra A. Covarrubias
  • Fernando Bastarrachea
Article

Summary

Mutants resistant to 80 μM L-methionine-DL-sulfoximine (MS) were isolated on glucose-minimal 15 mM NH + 4 medium plates from Escherichia coli cells which were hypersensitive to this concentration of the analogue by virtue of their harboring glnG mutations. MS-resistant mutants derived from strain MX902 carried, in addition to its glnG74::Tn5 allele, mutations tightly linked to glnA, as shown by P1-mediated transduction experiments. One particular allele, gln-76, which suppressed the MS-sensitivity conferred by glnG74::Tn5 but not its Ntr phenotype (inability to transport and utilize compounds such as arginine or proline as the only nitrogen sources), was shown to allow constitutive expression of glutamine synthetase in the absence not only of a functional glnG product but also of a functional glnF product. This behavior was found to be cis-dominant in complementation experiments with F'14 merogenotes. In an otherwise wild-type genetic background as in MX929 (gln-76 glnA+ glnL+ glnG+ glnF+), however, normal activation, mediated by the glnG and glnF products was preferred over that mediated by gln-76.

Keywords

Nitrogen Escherichia Coli Proline Arginine Glutamine 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Ausubel FM, Margolskee RF, Maizels N (1976) Mutants of Klebsiella pneumoniae in which expression of nitrogenase is independent of glutamine synthetase control. In: Newton W, Postgate JR, Rodriguez-Barrueco C (eds) Recent developments in nitrogen fixation. Academic Press, LOndon, pp 347–356Google Scholar
  2. Ausubel FM, Reidel G, Cannon F, Peskin A, Margolskee R (1977) Cloning nitrogen fixation genes from Klebsiella pneumoniae in vitro and the isolation of nif promoter mutants affecting glutamine synthetase regulation. In: Hollaender A (ed) Genetic engineering for nitrogen fixation. Plenum Press, New York, pp 111–128Google Scholar
  3. Ayling PD, Bridgeland ES (1972) Methionine transport in wild type and transport defective mutants of Salmonella typhimurium. J Gen Microbiol 73:127–141PubMedGoogle Scholar
  4. Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K-12, edition 6. Microbiol Revs 44:1–56Google Scholar
  5. Backman K, Chen Y, Magasanik B (1981) Physical and genetic characterization of the glnA-glnG region of the Escherichia coli chromosome. Proc Natl Acad Sci USA 78:3743–3747PubMedGoogle Scholar
  6. Bastarrachea F, Willetts NS (1968) The elimination by acridine orange of F30 from recombination-deficient strans of Escherichia coli K12. Genetics 59:153–166PubMedGoogle Scholar
  7. Bastarrachea F, Brom S, Covarrubias AA, Osorio A, Bolivar F (1980) Genetic characterization of mutations affecting glutamine biosynthesis and its regulation in Escherichia coli K12. In: Mora J, Palacios R (eds) Glutamine: Metabolsim, enzymology and regulation. Academic Press, New York, pp 107–121Google Scholar
  8. Berg DE, Davies J, Allet B, Rochaix J (1975) Transposition of R factor genes to bacteriophage λ. Proc Natl Acad Sci USA 72:3628–3632PubMedGoogle Scholar
  9. Betlach M, Hershfield V, Chow L, Brown W, Goodman HW, Boyer HW (1976) A restriction endonuclease analysis of the bacterial plasmid controlling the EcoRI restriction and modification of DNA. Fed Proc 35:2037–2043PubMedGoogle Scholar
  10. Betteridge PR, Ayling PD (1975) The role of methionine transportdefective mutations in resistance to methionine sulphoximine in Salmonella typhimurium. Mol Gen Genet 138:41–52CrossRefGoogle Scholar
  11. Chen Y, Backman K, Magasanik B (1982) Characterization of a gene, glnL, the product of which is involved in the regulation of nitrogen utilizationin Escherichia coli. J Bacteriol 150:214–220PubMedGoogle Scholar
  12. Covarrubias AA, Sanchez-Pescador R, Osorio A, Bolivar F, Bastarrachea F (1980) Col E1 hybrid plasmids containing Escherichia coli genes involved in the biosynthesis of glutamate and glutamine. Plasmid 3:150–164PubMedGoogle Scholar
  13. Covarrubias AA, Bastarrachea F (1983) Nucleotide sequence of the glnA control region of Escherichia coli. Mol Gen Genet 190:171–175PubMedGoogle Scholar
  14. De Lucia P, Cairns J (1969) Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature 224:1164–1166PubMedGoogle Scholar
  15. Garcia E, Bancroft S, Rhee SG, Kustu S (1977) The product of a newly identified gene, glnF, is required for synthesis of glutamine synthetase in Salmonella. Proc Natl Acad Sci USA 74:1662–1666PubMedGoogle Scholar
  16. Goldie H, Magasanik B (1982) Effects of glnL and other regulatory loci on regulation of transcription of glnA-lacZ fusions in Klebsiella aerogenes. J Bacteriol 150:231–238PubMedGoogle Scholar
  17. Greene PJ, Heyneker HL, Bolivar F, Rodriguez RL, Betlach MC, Covarrubias AA, Backman K, Russel DJ, Tait R, Boyer HW (1978) A general method for the purification of restriction enzymes. Nucleic Acids Res 5:2373–2380PubMedGoogle Scholar
  18. Guterman SK, Roberts G, Tyler B (1982) Polarity in the glnA operon: suppression of the Reg phenotype by rho mutations. J Bacteriol 150:1314–1321PubMedGoogle Scholar
  19. Kustu S, Burton D, Garcia E, McCarter L, MacFarland N (1979a) Nitrogen Control in Salmonella: regulation by the glnR and glnF gene products. Proc Natl Acad Sci USA 76:4576–4580PubMedGoogle Scholar
  20. Kustu SG, McFarland NC, Hui SP, Esmon B, Ames GF (1979b) Nitrogen control in Salmonella typhimurium: co-regulation of synthesis of glutamine synthetase and amino acid transport systems. J Bacteriol 138:218–234PubMedGoogle Scholar
  21. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  22. MacNeil D (1981) General method, using Mu-Mudl dilysogens, to determine the direction of transcription of and generate deletions in the glnA region of Escherichia coli. J Bacteriol 146:260–268PubMedGoogle Scholar
  23. MacNeil T, MacNeil D, Tyler B (1982) Fine-structure deletion map and complementation analysis of the glnA-glnL-glnG region of Escherichia coli. J Bacteriol 150:1302–1313PubMedGoogle Scholar
  24. Magasanik B (1982) Genetic control of nitrogen assimilation i bacteria. Annu Rev Genet 16:135–168CrossRefPubMedGoogle Scholar
  25. Manning JM, Moore S, Rowe WB, Meister A (1969) Identification of L-methionine S-sulfoximine as the diatercoisomer of L-methionine SR-sulfoximine that inhibits glutamine synthetase. Biochemistry 8:2681–2685PubMedGoogle Scholar
  26. Marmur J (1961) A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J Mol Biol 3:208–218Google Scholar
  27. McFarland N, McCarter L, Artz S, Kustu S (1981) Nitrogen regulatory locus “glnR” of enteric bacteria is composed by cistrons ntrB and ntrC: identification of their protein products. Proc Natl Acad Sci USA 78:2135–2139PubMedGoogle Scholar
  28. Miller ES, Brenchley JE (1981) L-methionine SR-sulfoximine-resistant glutamine synthetase from mutants of Salmonella typhimurium. J Biol Chem 256:11307–11312PubMedGoogle Scholar
  29. Miller J (1972) Experiments in molecular genetics. 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New YorkGoogle Scholar
  30. Pahel G, Zelenetz AD, Tyler BM (1978) gltB gene and regulation of nitrogen metabolism by glutamine synthetase in Escherichia coli. J Bacteriol 133:139–148PubMedGoogle Scholar
  31. Pahel G, Tyler B (1979) A new glnA-linked regulatory gene for glutamine synthetase in Escherichia coli. Proc Natl Acad Sci USA 76:4544–4548PubMedGoogle Scholar
  32. Pahel G, Rothstein DM, Magasanik B (1982) Complex glnA-glnL-glnG operon of Escherichia coli. J Bacteriol 150:202–213PubMedGoogle Scholar
  33. Ronzio RA, Meister A (1968) Phosphorylation of methionine sulfoximine by glutamine synthetase. Proc Natl Acad Sci USA 59:164–170PubMedGoogle Scholar
  34. Ronzio RA, Rowe WB, Meister A (1969) Studies on the mechanisms of inhibition of glutamine synthetase by methionine sulfoximine. Biochemistry 8:1066–1075PubMedGoogle Scholar
  35. Rothstein DM, Magasanik B (1980) Isolation of Klebsiella aerogenes mutants cis-dominant for glutamine synthetase expression. J Bacteriol 141:671–679PubMedGoogle Scholar
  36. Rothstein DM, Pahel G, Tyler B, Magasanik B (1980) Regulation of expression from the glnA promoter of Escherichia coli in the absence of glutamine synthetase. Proc Natl Acad Sci USA 77:7372–7376PubMedGoogle Scholar
  37. Rowe WB, Ronzio RA, Meister A (1969) Inhibition of glutamine synthetase by methionine sulfoximine. Studies on methionine sulfoximine phosphate. Biochemistry 8:2674–2680PubMedGoogle Scholar
  38. Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517PubMedGoogle Scholar
  39. Wei GR, Kustu S (1981) Glutamine auxotrophs with mutations in a nitrogen regulatory gene, ntrC, that is near glnA. Mol Gen Genet 181:392–399CrossRefGoogle Scholar
  40. Woolfolk CA, Shapiro BM, Stadtman ER (1966) Regulation of glutamine synthetase, I. Purification and properties of glutamine synthetase from Escherichia coli. Arch Biochem Biophys 116:177–192PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Aurora V. Osorio
    • 1
  • Luis Servín-González
    • 1
  • Mario Rocha
    • 1
  • Alejandra A. Covarrubias
    • 1
  • Fernando Bastarrachea
    • 1
  1. 1.Centro de Investigación sobre Ingeniería Genética y BiotecnologíaUniversidad Nacional Autónoma de MéxicoMéxico 20México

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