Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Jun;179(11):3786–3789. doi: 10.1128/jb.179.11.3786-3789.1997

Membrane topology of the metal-tetracycline/H+ antiporter TetA(K) from Staphylococcus aureus.

S L Ginn 1, M H Brown 1, R A Skurray 1
PMCID: PMC179179  PMID: 9171431

Abstract

A series of fusions to the reporter proteins alkaline phosphatase and beta-galactosidase have been constructed in the predicted periplasmic and cytoplasmic loops of TetA(K), a protein responsible for efflux-mediated tetracycline resistance in Staphylococcus aureus. The results support a topological model of 14 transmembrane segments for TetA(K).

Full Text

The Full Text of this article is available as a PDF (200.3 KB).

Selected References

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

  1. Allard J. D., Bertrand K. P. Membrane topology of the pBR322 tetracycline resistance protein. TetA-PhoA gene fusions and implications for the mechanism of TetA membrane insertion. J Biol Chem. 1992 Sep 5;267(25):17809–17819. [PubMed] [Google Scholar]
  2. Brickman E., Beckwith J. Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and phi80 transducing phages. J Mol Biol. 1975 Aug 5;96(2):307–316. doi: 10.1016/0022-2836(75)90350-2. [DOI] [PubMed] [Google Scholar]
  3. Claros M. G., von Heijne G. TopPred II: an improved software for membrane protein structure predictions. Comput Appl Biosci. 1994 Dec;10(6):685–686. doi: 10.1093/bioinformatics/10.6.685. [DOI] [PubMed] [Google Scholar]
  4. Eckert B., Beck C. F. Topology of the transposon Tn10-encoded tetracycline resistance protein within the inner membrane of Escherichia coli. J Biol Chem. 1989 Jul 15;264(20):11663–11670. [PubMed] [Google Scholar]
  5. Engelman D. M., Steitz T. A., Goldman A. Identifying nonpolar transbilayer helices in amino acid sequences of membrane proteins. Annu Rev Biophys Biophys Chem. 1986;15:321–353. doi: 10.1146/annurev.bb.15.060186.001541. [DOI] [PubMed] [Google Scholar]
  6. Fujihira E., Kimura T., Shiina Y., Yamaguchi A. Transmembrane glutamic acid residues play essential roles in the metal-tetracycline/H+ antiporter of Staphylococcus aureus. FEBS Lett. 1996 Aug 12;391(3):243–246. doi: 10.1016/0014-5793(96)00743-0. [DOI] [PubMed] [Google Scholar]
  7. Griffith J. K., Baker M. E., Rouch D. A., Page M. G., Skurray R. A., Paulsen I. T., Chater K. F., Baldwin S. A., Henderson P. J. Membrane transport proteins: implications of sequence comparisons. Curr Opin Cell Biol. 1992 Aug;4(4):684–695. doi: 10.1016/0955-0674(92)90090-y. [DOI] [PubMed] [Google Scholar]
  8. Guay G. G., Khan S. A., Rothstein D. M. The tet(K) gene of plasmid pT181 of Staphylococcus aureus encodes an efflux protein that contains 14 transmembrane helices. Plasmid. 1993 Sep;30(2):163–166. doi: 10.1006/plas.1993.1045. [DOI] [PubMed] [Google Scholar]
  9. Hoshino T., Ikeda T., Tomizuka N., Furukawa K. Nucleotide sequence of the tetracycline resistance gene of pTHT15, a thermophilic Bacillus plasmid: comparison with staphylococcal TcR controls. Gene. 1985;37(1-3):131–138. doi: 10.1016/0378-1119(85)90265-3. [DOI] [PubMed] [Google Scholar]
  10. Iordănescu S. Three distinct plasmids originating in the same Staphylococcus aureus strain. Arch Roum Pathol Exp Microbiol. 1976 Jan-Jun;35(1-2):111–118. [PubMed] [Google Scholar]
  11. Kunkel T. A., Roberts J. D., Zakour R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods Enzymol. 1987;154:367–382. doi: 10.1016/0076-6879(87)54085-x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Levy S. B. Active efflux mechanisms for antimicrobial resistance. Antimicrob Agents Chemother. 1992 Apr;36(4):695–703. doi: 10.1128/aac.36.4.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Manoil C., Beckwith J. TnphoA: a transposon probe for protein export signals. Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129–8133. doi: 10.1073/pnas.82.23.8129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marger M. D., Saier M. H., Jr A major superfamily of transmembrane facilitators that catalyse uniport, symport and antiport. Trends Biochem Sci. 1993 Jan;18(1):13–20. doi: 10.1016/0968-0004(93)90081-w. [DOI] [PubMed] [Google Scholar]
  16. Michaelis S., Inouye H., Oliver D., Beckwith J. Mutations that alter the signal sequence of alkaline phosphatase in Escherichia coli. J Bacteriol. 1983 Apr;154(1):366–374. doi: 10.1128/jb.154.1.366-374.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Paulsen I. T., Brown M. H., Dunstan S. J., Skurray R. A. Molecular characterization of the staphylococcal multidrug resistance export protein QacC. J Bacteriol. 1995 May;177(10):2827–2833. doi: 10.1128/jb.177.10.2827-2833.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Paulsen I. T., Brown M. H., Littlejohn T. G., Mitchell B. A., Skurray R. A. Multidrug resistance proteins QacA and QacB from Staphylococcus aureus: membrane topology and identification of residues involved in substrate specificity. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3630–3635. doi: 10.1073/pnas.93.8.3630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Paulsen I. T., Brown M. H., Skurray R. A. Proton-dependent multidrug efflux systems. Microbiol Rev. 1996 Dec;60(4):575–608. doi: 10.1128/mr.60.4.575-608.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Paulsen I. T., Skurray R. A. Topology, structure and evolution of two families of proteins involved in antibiotic and antiseptic resistance in eukaryotes and prokaryotes--an analysis. Gene. 1993 Feb 14;124(1):1–11. doi: 10.1016/0378-1119(93)90755-r. [DOI] [PubMed] [Google Scholar]
  21. Rouch D. A., Cram D. S., DiBerardino D., Littlejohn T. G., Skurray R. A. Efflux-mediated antiseptic resistance gene qacA from Staphylococcus aureus: common ancestry with tetracycline- and sugar-transport proteins. Mol Microbiol. 1990 Dec;4(12):2051–2062. doi: 10.1111/j.1365-2958.1990.tb00565.x. [DOI] [PubMed] [Google Scholar]
  22. Smith R. L., Banks J. L., Snavely M. D., Maguire M. E. Sequence and topology of the CorA magnesium transport systems of Salmonella typhimurium and Escherichia coli. Identification of a new class of transport protein. J Biol Chem. 1993 Jul 5;268(19):14071–14080. [PubMed] [Google Scholar]
  23. Traxler B., Boyd D., Beckwith J. The topological analysis of integral cytoplasmic membrane proteins. J Membr Biol. 1993 Feb;132(1):1–11. doi: 10.1007/BF00233047. [DOI] [PubMed] [Google Scholar]
  24. Vieira J., Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. doi: 10.1016/0076-6879(87)53044-0. [DOI] [PubMed] [Google Scholar]
  25. Yamaguchi A., Shiina Y., Fujihira E., Sawai T., Noguchi N., Sasatsu M. The tetracycline efflux protein encoded by the tet(K) gene from Staphylococcus aureus is a metal-tetracycline/H+ antiporter. FEBS Lett. 1995 May 29;365(2-3):193–197. doi: 10.1016/0014-5793(95)00455-i. [DOI] [PubMed] [Google Scholar]
  26. von Heijne G. Membrane proteins: the amino acid composition of membrane-penetrating segments. Eur J Biochem. 1981 Nov;120(2):275–278. doi: 10.1111/j.1432-1033.1981.tb05700.x. [DOI] [PubMed] [Google Scholar]

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

RESOURCES