Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Fluorocitrate resistant tricarboxylate transport mutants of Salmonella typhimurium

Summary

Spontaneous and Tn10 induced fluorocitrate resistant mutants were isolated and characterized. These mutants were unable to grow on either cis-aconitate or DL-isocitrate but were still able to grow slowly on sodium citrate and normally on potassium or potassium-plus-sodium citrate. These mutants were defective in both citrate transport and citrate binding to periplasmic proteins. Tn10 insertion mutants were unable to produce immunologically detectable amounts of the citrate inducible periplasmic C protein previously shown to bind tricarboxylates.

Using a series of tct::Tn10 directed Hfrs the tct locus was accurately positioned at 59 units between srlA and pheA, but was not cotransducible with either gene. In the absence of P22 mediated cotransduction with 16 adjacent chromosomal markers the srlA and tct loci were bridged by using a series of tct flanking Tn10 insertions, and by newly isolated and characterized nalB mutants. In addition the hyd and recA loci were located establishing the gene order in this region of the chromosome as: pheA tct nalB recA srlA hyd cys. Nitrosoguanidine derived tricarboxylate mutations (Imai 1975) were also mapped within the tct locus.

This is a preview of subscription content, log in to check access.

References

  1. Abou-Jaoude A, Pascal MC, Casse F, Chippaux M (1978) Isolation and phenotypes of mutants from Escherichia coli K12 defective in nitrite reductase activity. FEMS Microbiol Lett 3:235–239

  2. Ames GF (1974) Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem 249:634–644

  3. Ashton DM, Sweet GD, Somers JM, Kay WW (1980) Citrate transport in Salmonella typhimurium: studies with 2-fluoro-L-erythro-citrate as a substrate. Can J Biochem 58:797–803

  4. Chan RK, Botstein D, Watanabe T, Ogata Y (1972) Specialized transduction of tetracycline resistance by phage P22 in Salmonella typhimurium. II. Properties of a high-frequency-transducing lysate. Virology 50:883–898

  5. Chumley FG, Menzel R, Roth JR (1979) Hfr formation directed by Tn10. Genetics 91, 639–655

  6. Frost GE, Rosenberg H (1973) The citrate-dependent iron transport system in Escherichia coli K-12. Biochim Biophys. Acta 330:90–101

  7. Hane MW, Wood TH (1969) Escherichia coli mutants resistant to nalidixic acid: genetic mapping and dominance studies. J Bacteriol 99:238–241

  8. Hancock REW, Hantke K, Braun V (1976) Iron transport in Escherichia coli K12: involvement of the colicin B receptor and of a citrate-inducible protein. J Bacteriol 127:1370–1375

  9. Imai K (1975) Isolation of tricarboxylate transport-negative mutants of Salmonella typhimurium. J Gen Appl Microbiol 21:127–134

  10. Imai K, Iijima T, Banno I (1977) Location of tct (tricarboxylic acid transport) genes on the chromosome of Salmonella typhimurium. Inst Ferment Res Commun (Osaka) 8:63–68

  11. Ingolia TJ, Koshland Jr, DE (1979) Response to a metal ion citrate complex in bacterial sensing. J Bacteriol 140:798–804

  12. Kay WW (1978) Carboxylic acid transport. In: Rosen BP (ed) Bacterial transport. Marcel Dekker, Inc, New York, p 385

  13. Kay WW, Cameron M (1978) Citrate transport in Salmonella typhimurium. Arch Biochem Biophys 190:270–278

  14. Kemper J (1974) Gene order and co-transduction in the leu-ara-fob-pyrA region of Salmonella typhimurium linkage map. J Bacteriol 117:94–99

  15. Kihara M, Macnab RM (1979) Chemotaxis of Salmonella typhimurium toward citrate. J Bacteriol 140:297–300

  16. Kleckner N, Chan RK, Tye B, Botstein D (1975) Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J Mol Biol 97:561–575

  17. Kleckner N, Roth J, Botstein D (1977) Genetic engineering in vivo using translocatable drug-resistance elements. New methods in bacterial genetics. J Mol Biol 116:125–159

  18. Kleckner N, Reichardt K, Botstein D (1979) Inversions and deletions of the Salmonella chromosome generated by the translocatable tetracycline resistance element Tn10. J Mol Biol 127:89–115

  19. Sanderson KE, Hartman P (1978) Linkage map of Salmonella typhimurium. Microbiol Rev 42:471–519

  20. Sweet GD, Somers JM, Kay WW (1979) Purification and properties of a citrate-binding transport component, the C protein of Salmonella typhimurium. Can J Biochem 57:710–715

  21. Willis RE, Morris RG, Cirakoglu C, Schellenerg GD, Gerber NH, Furlong CE (1975) Preparation of the periplasmic binding proteins from Salmonella typhimurium and Escherichia coli. Arch Biochem Biophys 161:64–75

Download references

Author information

Correspondence to W. W. Kay.

Additional information

Communicated by G.O'Donovan

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Somers, J.M., Sweet, G.D. & Kay, W.W. Fluorocitrate resistant tricarboxylate transport mutants of Salmonella typhimurium . Molec. Gen. Genet. 181, 338–345 (1981). https://doi.org/10.1007/BF00425608

Download citation

Keywords

  • Sodium
  • Potassium
  • Citrate
  • Sodium Citrate
  • Gene Order