Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1982 Aug;151(2):620–628. doi: 10.1128/jb.151.2.620-628.1982

Mutational separation of transport systems for branched-chain amino acids in Pseudomonas aeruginosa.

T Hoshino, M Kageyama
PMCID: PMC220302  PMID: 6807957

Abstract

Several types of Pseudomonas aeruginosa mutants defective in the transport systems for branched-chain amino acids were isolated by selection for resistance to 5',5',5'-DL-trifluoroleucine, a leucine analog, under certain conditions. Mutants resistant to trifluoroleucine in the absence of Na+ were defective in the high-affinity system. These mutants fell into two classes. One class showed a defect in the production of a periplasmic binding protein for leucine, isoleucine, valine, alanine, and threonine, and the other showed normal production of the binding protein as determined by a binding assay and by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Properties of the former class of mutants have been partly described (T. Hoshino and M. Kageyama, J. Bacteriol. 141:1055-1063, 1980). Mutants selected for resistance to trifluoroleucine with Na+ and an excess amount of alanine showed a defect in the low-affinity system. Membrane vesicles prepared from such a mutant lost the transport activity for leucine. A mutant which showed increased activity of the low-affinity system with a defect in the high-affinity system was obtained from strain PML1453 (high-affinity system defective) by selecting for utilization of isoleucine as a carbon source.

Full text

PDF
622

Images in this article

624Image
on p.624

Selected References

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

  1. Ames G. F., Roth J. R. Histidine and aromatic permeases of Salmonella typhimurim. J Bacteriol. 1968 Nov;96(5):1742–1749. doi: 10.1128/jb.96.5.1742-1749.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. J., Oxender D. L. Escherichia coli transport mutants lacking binding protein and other components of the branched-chain amino acid transport systems. J Bacteriol. 1977 Apr;130(1):384–392. doi: 10.1128/jb.130.1.384-392.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson J. J., Oxender D. L. Genetic separation of high- and low-affinity transport systems for branched-chain amino acids in Escherichia coli K-12. J Bacteriol. 1978 Oct;136(1):168–174. doi: 10.1128/jb.136.1.168-174.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anraku Y. Transport of sugars and amino acids in bacteria. I. Purification and specificity of the galactose- and leucine-binding proteins. J Biol Chem. 1968 Jun 10;243(11):3116–3122. [PubMed] [Google Scholar]
  5. FARGIE B., HOLLOWAY B. W. ABSENCE OF CLUSTERING OF FUNCTIONALLY RELATED GENES IN PSEUDOMONAS AERUGINOSA. Genet Res. 1965 Jul;6:284–299. doi: 10.1017/s0016672300004158. [DOI] [PubMed] [Google Scholar]
  6. Guardiola J., De Felice M., Klopotowski T., Iaccarino M. Mutations affecting the different transport systems for isoleucine, leucine, and valine in Escherichia coli K-12. J Bacteriol. 1974 Feb;117(2):393–405. doi: 10.1128/jb.117.2.393-405.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guardiola J., Iaccarino M. Escherichia coli K-12 mutants altered in the transport of branched-chain amino acids. J Bacteriol. 1971 Dec;108(3):1034–1044. doi: 10.1128/jb.108.3.1034-1044.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hoshino T., Kageyama M. Purification and properties of a binding protein for branched-chain amino acids in Pseudomonas aeruginosa. J Bacteriol. 1980 Mar;141(3):1055–1063. doi: 10.1128/jb.141.3.1055-1063.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hoshino T., Kageyama M. Sodium-dependent transport of L-leucine in membrane vesicles prepared from Pseudomonas aeruginosa. J Bacteriol. 1979 Jan;137(1):73–81. doi: 10.1128/jb.137.1.73-81.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hoshino T. Transport systems for branched-chain amino acids in Pseudomonas aeruginosa. J Bacteriol. 1979 Sep;139(3):705–712. doi: 10.1128/jb.139.3.705-712.1979. [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. LUBIN M., KESSEL D. H., BUDREAU A., GROSS J. D. The isolation of bacterial mutants defective in amino acid transport. Biochim Biophys Acta. 1960 Aug 26;42:535–538. doi: 10.1016/0006-3002(60)90836-2. [DOI] [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Landick R., Anderson J. J., Mayo M. M., Gunsalus R. P., Mavromara P., Daniels C. J., Oxender D. L. Regulation of high-affinity leucine transport in Escherichia coli. J Supramol Struct. 1980;14(4):527–537. doi: 10.1002/jss.400140410. [DOI] [PubMed] [Google Scholar]
  15. Marinus M. G., Loutit J. S. Regulation of isoleucine-valine biosynthesis in Pseudomonas aeruginosa. II. Regulation of enzyme activity and synthesis. Genetics. 1969 Nov;63(3):557–567. doi: 10.1093/genetics/63.3.557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mizuno T., Kageyama M. Separation and characterization of the outer membrane of Pseudomonas aeruginosa. J Biochem. 1978 Jul;84(1):179–191. doi: 10.1093/oxfordjournals.jbchem.a132106. [DOI] [PubMed] [Google Scholar]
  17. Motojima K., Yamato I., Anraku Y. Proline transport carrier-defective mutants of Escherichia coli K-12: properties and mapping. J Bacteriol. 1978 Oct;136(1):5–9. doi: 10.1128/jb.136.1.5-9.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Neal J. L. Analysis of Michaelis kinetics for two independent, saturable membrane transport functions. J Theor Biol. 1972 Apr;35(1):113–118. doi: 10.1016/0022-5193(72)90196-8. [DOI] [PubMed] [Google Scholar]
  19. Oxender D. L., Anderson J. J., Daniels C. J., Landick R., Gunsalus R. P., Zurawski G., Selker E., Yanofsky C. Structural and functional analysis of cloned DNA containing genes responsible for branched-chain amino acid transport in Escherichia coli. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1412–1416. doi: 10.1073/pnas.77.3.1412. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rosen B. P. Basic amino acid transport in Escherichia coli: properties of canavanine-resistant mutants. J Bacteriol. 1973 Nov;116(2):627–635. doi: 10.1128/jb.116.2.627-635.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yamato I., Anraku Y. Genetic and biochemical studies of transport systems for branched-chain amino acids in Escherichia coli K-12: isolation and properties of mutants defective in leucine-repressible transport activities. J Bacteriol. 1980 Oct;144(1):36–44. doi: 10.1128/jb.144.1.36-44.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yamato I., Ohki M., Anraku Y. Genetic and biochemical studies of transport systems for branched-chain amino acids in Escherichia coli. J Bacteriol. 1979 Apr;138(1):24–32. doi: 10.1128/jb.138.1.24-32.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES