Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1995 Jan;61(1):252–259. doi: 10.1128/aem.61.1.252-259.1995

Identification of a PutP proline permease gene homolog from Staphylococcus aureus by expression cloning of the high-affinity proline transport system in Escherichia coli.

P A Wengender 1, K J Miller 1
PMCID: PMC167280  PMID: 7887605

Abstract

The important food-borne pathogen Staphylococcus aureus is distinguished by its ability to grow at low water activity values. Previous work in our laboratory and by others has revealed that proline accumulation via transport is an important osmoregulatory strategy employed by this bacterium. Furthermore, proline uptake by this bacterium has been shown to be mediated by two distinct transport systems: a high-affinity system and a low-affinity system (J.-H. Bae, and K. J. Miller, Appl. Environ. Microbiol. 58:471-475, 1992; D. E. Townsend and B. J. Wilkinson, J. Bacteriol. 174:2702-2710, 1992). In the present study, we report the cloning of the high-affinity proline transport system of S. aureus by functional expression in an Escherichia coli host. The sequence of the staphylococcal proline permease gene was predicted to encode a protein of 497 amino acids which shares 49% identity with the PutP high-affinity proline permease of E. coli. Analysis of hydropathy also indicated a common overall structure for these proteins.

Full Text

The Full Text of this article is available as a PDF (295.9 KB).

Selected References

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

  1. Altschul S. F., Gish W., Miller W., Myers E. W., Lipman D. J. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. doi: 10.1016/S0022-2836(05)80360-2. [DOI] [PubMed] [Google Scholar]
  2. Amanuma H., Itoh J., Anraku Y. Proton-dependent binding of proline to carrier in Escherichia coli membrane. FEBS Lett. 1977 Jun 15;78(2):173–176. doi: 10.1016/0014-5793(77)80299-8. [DOI] [PubMed] [Google Scholar]
  3. Bae J. H., Anderson S. H., Miller K. J. Identification of a high-affinity glycine betaine transport system in Staphylococcus aureus. Appl Environ Microbiol. 1993 Aug;59(8):2734–2736. doi: 10.1128/aem.59.8.2734-2736.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bae J. H., Miller K. J. Identification of two proline transport systems in Staphylococcus aureus and their possible roles in osmoregulation. Appl Environ Microbiol. 1992 Feb;58(2):471–475. doi: 10.1128/aem.58.2.471-475.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breidt F., Jr, Hengstenberg W., Finkeldei U., Stewart G. C. Identification of the genes for the lactose-specific components of the phosphotransferase system in the lac operon of Staphylococcus aureus. J Biol Chem. 1987 Dec 5;262(34):16444–16449. [PubMed] [Google Scholar]
  6. Capaldi R. A., Vanderkooi G. The low polarity of many membrane proteins. Proc Natl Acad Sci U S A. 1972 Apr;69(4):930–932. doi: 10.1073/pnas.69.4.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Csonka L. N. Physiological and genetic responses of bacteria to osmotic stress. Microbiol Rev. 1989 Mar;53(1):121–147. doi: 10.1128/mr.53.1.121-147.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Culham D. E., Lasby B., Marangoni A. G., Milner J. L., Steer B. A., van Nues R. W., Wood J. M. Isolation and sequencing of Escherichia coli gene proP reveals unusual structural features of the osmoregulatory proline/betaine transporter, ProP. J Mol Biol. 1993 Jan 5;229(1):268–276. doi: 10.1006/jmbi.1993.1030. [DOI] [PubMed] [Google Scholar]
  9. Deguchi Y., Yamato I., Anraku Y. Nucleotide sequence of gltS, the Na+/glutamate symport carrier gene of Escherichia coli B. J Biol Chem. 1990 Dec 15;265(35):21704–21708. [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dila D. K., Maloy S. R. Proline transport in Salmonella typhimurium: putP permease mutants with altered substrate specificity. J Bacteriol. 1986 Nov;168(2):590–594. doi: 10.1128/jb.168.2.590-594.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Dyer D. W., Iandolo J. J. Rapid isolation of DNA from Staphylococcus aureus. Appl Environ Microbiol. 1983 Jul;46(1):283–285. doi: 10.1128/aem.46.1.283-285.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Faatz E., Middendorf A., Bremer E. Cloned structural genes for the osmotically regulated binding-protein-dependent glycine betaine transport system (ProU) of Escherichia coli K-12. Mol Microbiol. 1988 Mar;2(2):265–279. doi: 10.1111/j.1365-2958.1988.tb00028.x. [DOI] [PubMed] [Google Scholar]
  14. Graham J. E., Wilkinson B. J. Staphylococcus aureus osmoregulation: roles for choline, glycine betaine, proline, and taurine. J Bacteriol. 1992 Apr;174(8):2711–2716. doi: 10.1128/jb.174.8.2711-2716.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grothe S., Krogsrud R. L., McClellan D. J., Milner J. L., Wood J. M. Proline transport and osmotic stress response in Escherichia coli K-12. J Bacteriol. 1986 Apr;166(1):253–259. doi: 10.1128/jb.166.1.253-259.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hanada K., Yoshida T., Yamato I., Anraku Y. Sodium ion and proline binding sites in the Na+/proline symport carrier of Escherichia coli. Biochim Biophys Acta. 1992 Mar 23;1105(1):61–66. doi: 10.1016/0005-2736(92)90162-f. [DOI] [PubMed] [Google Scholar]
  17. Hediger M. A., Turk E., Wright E. M. Homology of the human intestinal Na+/glucose and Escherichia coli Na+/proline cotransporters. Proc Natl Acad Sci U S A. 1989 Aug;86(15):5748–5752. doi: 10.1073/pnas.86.15.5748. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kaback H. R., Patel L. The role of functional sulfhydryl groups in active transport in Escherichia coli membrane vesicles. Biochemistry. 1978 May 2;17(9):1640–1646. doi: 10.1021/bi00602a010. [DOI] [PubMed] [Google Scholar]
  19. Kunin C. M., Rudy J. Effect of NaCl-induced osmotic stress on intracellular concentrations of glycine betaine and potassium in Escherichia coli, Enterococcus faecalis, and staphylococci. J Lab Clin Med. 1991 Sep;118(3):217–224. [PubMed] [Google Scholar]
  20. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  21. Miller K., Maloy S. DNA sequence of the putP gene from Salmonella typhimurium and predicted structure of proline permease. Nucleic Acids Res. 1990 May 25;18(10):3057–3057. doi: 10.1093/nar/18.10.3057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Myers R. S., Maloy S. R. Mutations of putP that alter the lithium sensitivity of Salmonella typhimurium. Mol Microbiol. 1988 Nov;2(6):749–755. doi: 10.1111/j.1365-2958.1988.tb00086.x. [DOI] [PubMed] [Google Scholar]
  23. Nakao T., Yamato I., Anraku Y. Nucleotide sequence of putP, the proline carrier gene of Escherichia coli K12. Mol Gen Genet. 1987 Jun;208(1-2):70–75. doi: 10.1007/BF00330424. [DOI] [PubMed] [Google Scholar]
  24. Peerce B. E., Wright E. M. Evidence for tyrosyl residues at the Na+ site on the intestinal Na+/glucose cotransporter. J Biol Chem. 1985 May 25;260(10):6026–6031. [PubMed] [Google Scholar]
  25. Pourkomailian B., Booth I. R. Glycine betaine transport by Staphylococcus aureus: evidence for two transport systems and for their possible roles in osmoregulation. J Gen Microbiol. 1992 Dec;138(12):2515–2518. doi: 10.1099/00221287-138-12-2515. [DOI] [PubMed] [Google Scholar]
  26. Reizer J., Reizer A., Saier M. H., Jr A functional superfamily of sodium/solute symporters. Biochim Biophys Acta. 1994 Jun 29;1197(2):133–166. doi: 10.1016/0304-4157(94)90003-5. [DOI] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Short S. A., White D. C., Kaback H. R. Mechanisms of active transport in isolated bacterial membrane vesicles. IX. The kinetics and specificity of amino acid transport in Staphylococcus aureus membrane vesicles. J Biol Chem. 1972 Dec 10;247(23):7452–7458. [PubMed] [Google Scholar]
  29. Stormo G. D., Schneider T. D., Gold L. M. Characterization of translational initiation sites in E. coli. Nucleic Acids Res. 1982 May 11;10(9):2971–2996. doi: 10.1093/nar/10.9.2971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Townsend D. E., Wilkinson B. J. Proline transport in Staphylococcus aureus: a high-affinity system and a low-affinity system involved in osmoregulation. J Bacteriol. 1992 Apr;174(8):2702–2710. doi: 10.1128/jb.174.8.2702-2710.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wood J. M. Proline porters effect the utilization of proline as nutrient or osmoprotectant for bacteria. J Membr Biol. 1988 Dec;106(3):183–202. doi: 10.1007/BF01872157. [DOI] [PubMed] [Google Scholar]
  32. Yamato I., Anraku Y. Site-specific alteration of cysteine 281, cysteine 344, and cysteine 349 in the proline carrier of Escherichia coli. J Biol Chem. 1988 Nov 5;263(31):16055–16057. [PubMed] [Google Scholar]
  33. Yamato I., Kotani M., Oka Y., Anraku Y. Site-specific alteration of arginine 376, the unique positively charged amino acid residue in the mid-membrane-spanning regions of the proline carrier of Escherichia coli. J Biol Chem. 1994 Feb 25;269(8):5720–5724. [PubMed] [Google Scholar]
  34. Yamato I., Ohsawa M., Anraku Y. Defective cation-coupling mutants of Escherichia coli Na+/proline symport carrier. Characterization and localization of mutations. J Biol Chem. 1990 Feb 15;265(5):2450–2455. [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES