1887

Abstract

Summary: Wild-type expresses three proline transport systems: a high-affinity proline transport system encoded by the gene, and two glycine betaine transport systems with a lower affinity for proline encoded by the and genes. Although proline uptake by the ProP and ProU transport systems is sufficient to supplement a proline auxotroph, it is not efficient enough to allow proline utilization as a sole source of carbon or nitrogen. Thus, the PutP transport system is required for utilization of proline as a carbon or nitrogen source. In this study, an overexpression suppressor, designated , which allows proline utilization in a genetic background and does not require the function of any of the known proline transport systems, was cloned and characterized. The suppressor gene, designated , maps at 8 min on the linkage map, distant from any of the other characterized proline transport genes. The DNA sequence of the gene predicts that it encodes a hydrophobic integral membrane protein, with strong similarity to a family of amino acid transporters. The suppressor phenotype requires either a multicopy clone of the gene or both a single copy of the gene and a mutation. These results indicate that the gene is the structural gene for a cryptic proline transporter that is silent unless overexpressed on a multicopy plasmid or activated by a mutation.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-143-9-2903
1997-09-01
2021-07-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/143/9/mic-143-9-2903.html?itemId=/content/journal/micro/10.1099/00221287-143-9-2903&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. 1990; Basic local alignment search tool. J MolBiol 215:403–410
    [Google Scholar]
  2. Beatriz A., Olfson P., Casadaban M. 1984; Plasmid insertion mutagenesis and lac gene fusion with mini-Mu bacteriophage transposons. Proc Natl Acad Sci USA 158:488–495
    [Google Scholar]
  3. Begley G., Warner K., Arents J., Postma P., Jacobson G. 1996; Isolation and characterization of a mutation that alters the substrate specificity of the Escherichia coli glucose permease. J Bacteriol 178:940–942
    [Google Scholar]
  4. Biek D. P., Cohen S. N. 1986; Identification and characterization of recD, a gene affecting plasmid maintenance and recombination in Escherichia coli . J Bacteriol 167:594–603
    [Google Scholar]
  5. Bochner B. 1984; Curing bacterial cells of lysogenic viruses by using UCB indicator plates. BioTechniques 2:234–240
    [Google Scholar]
  6. Claros M., von Heijne G. 1994; Prediction of transmembrane segments in integral membrane proteins, and the putative topologies. CABIOS 10:685–686
    [Google Scholar]
  7. Covarrubias L., Cervantes L., Covarrubias A., Soberon X., Vichido I., Glanco A., Kupersztoch-Portnoy Y., Bolivar F. 1981; Construction of new cloning vehicles. V. Mobilization and coding properties of pBR322 and several deletion derivatives including pBR327 and pBR328. Gene 1325–35
    [Google Scholar]
  8. Csonka L. 1982; A third l-prolinepermease in Salmonella typhimurium which functions in media of elevated osmotic strength. J Bacteriol 151:1433–1443
    [Google Scholar]
  9. Csonka L., Epstein W. 1996; Osmoregulation. In Escherichia coli and Salmonella: Cellular and Molecular Biology 2nd edn, pp. 1210–1224 Edited by Neidhardt F. C. and others Washington, DC: American Society for Microbiology;
    [Google Scholar]
  10. Davis R., Botstein D., Roth R. J. 1980 Advanced Bacterial Genetics. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  11. Dila D., Maloy S. 1987; Proline transport in Salmonella typhimurium: putP permease mutants with altered substrate specificity. J Bacteriol 168:590–594
    [Google Scholar]
  12. Ekena K., Liao M.-K., Maloy S. 1990; Activation of a cryptic proline transport system in Salmonella typhimurium . J Bacteriol 172:2940–2945
    [Google Scholar]
  13. Elliott T., Roth J. R. 1988; Characterization of Tn10d-Cam: a transposition-defective Tn10 specifying chloramphenicol resistance. Mol Gen Genet 213:332–338
    [Google Scholar]
  14. Fsihi H., Kottwitz B., Gremer E. 1993; Single amino acid substitutions affecting the substrate specificity of the Escherichia coli K-12 nucleoside-specific Tsx channel. J BiolChem 268:17495–17503
    [Google Scholar]
  15. Giel M., Desnoyer M., Lopilato J. 1996; A mutation in a new gene, bglJ, activates thebgl operon in Escherichia coli K-12. Genetics 143:627–635
    [Google Scholar]
  16. Grothe S., Krogsrud R., McClellan D., Milner J., Wood J. 1986; Proline transport and osmotic stress in Escherichia coli K-12. J Bacteriol 166:253–259
    [Google Scholar]
  17. Hall B., Yokoyama S., Calhoun D. 1983; Role of cryptic genes in microbial evolution. Mol Biol Evol 1:109–124
    [Google Scholar]
  18. vonHeijne G. 1987 Sequence Analysis in Molecular Biology: Treasure Trove or Trivial Pursuit. San Diego, CA: Academic Press;
    [Google Scholar]
  19. Higgins C., Sutherland L., Cairney J., Booth I. 1987; Theosmotically regulated proU locus of Salmonella typhimuriumencodes a periplasmicbetaine-binding protein. J Gen Microbiol 133:305–310
    [Google Scholar]
  20. Hmiel S. P., Snavely M. D., Miller C. G., Maguire M. E. 1986; Magnesium transport in Salmonella typhimurium:characterization of magnesium influx and cloning of a transport gene. J Bacteriol 168:1444–1450
    [Google Scholar]
  21. Holloway B., Low K. 1996; F-prime and R-prime factors. In Escherichia coli and Salmonella: Cellular and Molecular Biology 2nd edn, pp. 1210–1224 Edited by Neidhardt F. C. and others Washington, DC: American Society for Microbiology;
    [Google Scholar]
  22. Hughes K. T., Roth J. R. 1985; Directed formation of deletions and duplications using Mud(Ap, lac). Genetics 109:263–282
    [Google Scholar]
  23. Hughes K. T., Roth J. R. 1988; Transitory cis complementation: a method for providing transposition functions to defective transposons. Genetics 119:9–12
    [Google Scholar]
  24. King S., Wilson T. 1990; Identification of valine 177 as a mutation altering specificity for transport of sugars by the Escherichia coli lactose carrier. Enhanced specificity for sucrose and maltose. J BiolChetn 265:9638–9644
    [Google Scholar]
  25. Kiritani K., Ohnishi K. 1987; Multiple transport systems for branched-chain amino acids as studied by mutants of Salmonella typhimurium . Jpn J Genet 53:265–274
    [Google Scholar]
  26. Kyte J., Doolittle R. 1982; A simple method for displaying the hydropathic character of a protein. J MolBiol 157:105–132
    [Google Scholar]
  27. Maloy S. 1989 Experimental Techniques in Bacterial Genetics. Boston: Jones and Bartlett;
    [Google Scholar]
  28. Maloy S., Roth J. 1983; Regulation of proline utilization in Salmonella typhimurium: characterization of put::Mud(Ap, lac) operon fusions. J Bacteriol 154:561–568
    [Google Scholar]
  29. Maloy S., Stewart V., Taylor R. 1996 Genetic Analysis of Pathogenic Bacteria. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  30. Matsubara K., Ohnishi K., Kiritani K. 1987; Location of livA gene participating in the high-affinity transport of branch-chained amino acids in Salmonella typhimurium LT2. Jpn J Genet 62:189–196
    [Google Scholar]
  31. Mead D., Szczesna-Sorupa E., Kemper B. 1986; Single-stranded DNA ‘Blue’ T7 promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng 1:67–74
    [Google Scholar]
  32. Menzel R., Roth J. 1980; Identification and mapping of a second prolinepermease in Salmonella typhimurium . J Bacteriol 141:1064–1070
    [Google Scholar]
  33. Miesel L., Roth J. 1994; Salmonella recD mutations increase recombination in a short sequence transduction assay. J Bacteriol 176:4092–4103
    [Google Scholar]
  34. Milner J. L., Grothe S., Wood J. M. 1988; Proline porter II is activated by a hyperosmotic shift in both whole cells and membrane vesicles of Escherichia coli K12. J BiolChem 263:14900–14905
    [Google Scholar]
  35. Myers R., Townsend D., Maloy S. 1991; Dissecting the molecular mechanism of ion-solute cotransport: substrate specificity mutations in the putP gene affect the kinetics of proline transport. J MembrBiol 121:201–214
    [Google Scholar]
  36. Ohnishi K., Hasegawa A., Matsubara K., Date T., Okada T., Kiritani K. 1988; Cloning and nucleotide sequence of the brnQ gene, the structural gene for a membrane-associated component of the LIV-II transport system for branched-chain amino acids in Salmonella typhimurium . Jpn J Genet 63:343–357
    [Google Scholar]
  37. Ratzkin B., Roth J. 1978; Cluster of genes controlling proline degradation in Salmonella typhimurium . J Bacteriol 133:733–744
    [Google Scholar]
  38. Russell C., Thaler D., Dahlquist F. 1989; Chromosomal transformation of Escherichia coli recD strains with linearized plasmids. J Bacteriol 171:2609–2613
    [Google Scholar]
  39. Sanderson K., Hessel A., Liu S.-L., Rudd K. 1996; The genetic linkage map of Salmonella typhimurium, edition VIII. In Escherichia coli and Salmonella: Cellular and Molecular Biology 2nd edn, pp. 1210–1224 Edited by Neidhardt F. C. and others Washington, DC: American Society for Microbiology;
    [Google Scholar]
  40. Stalmach M., Grothe S., Wood J. 1983; Two proline porters in Escherichia coli K-12. J Bacteriol 156:481–486
    [Google Scholar]
  41. Sturrock S. S., Collins J. F. 1993; MPsrch version 1.3. Biocomputing Research Unit, University of Edinburgh; UK:
    [Google Scholar]
  42. Vogel H. J., Bonner D. M. 1956; Acetylornithinase of Escherichia coli: partial purification and some properties. J BiolChem 218:97–106
    [Google Scholar]
  43. Wang A., Roth J. 1988; Activation of silent genes by transposons Tn5 and Tn10 . Genetics 120:875–885
    [Google Scholar]
  44. Wood J. M. 1988; Proline porters effect the utilization of proline as a nutrient or osmoprotectant for bacteria. J MembrBiol 106:183–202
    [Google Scholar]
  45. Wood J. M., Zadworny D. 1979; Characterization of an inducible porter required for l-proline catabolism by Escherichia coli K-12. Can J Biochem 57:1191–1199
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-143-9-2903
Loading
/content/journal/micro/10.1099/00221287-143-9-2903
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error