Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Sep;177(18):5355–5357. doi: 10.1128/jb.177.18.5355-5357.1995

Genetic analysis suggests functional interactions between the N- and C-terminal domains of the TetA(C) efflux pump encoded by pBR322.

P McNicholas 1, M McGlynn 1, G G Guay 1, D M Rothstein 1
PMCID: PMC177333  PMID: 7665527

Abstract

Genetic analysis of the tetA(C) gene of pBR322 indicates the importance of two-cytoplasmic loops in the TetA(C) protein (P. McNicholas, I. Chopra, and D. M. Rothstein, J. Bacteriol. 174:7926-7933, 1992). In this study, we characterized second-site suppressor mutations that suggest a functional interaction between these two cytoplasmic regions of the protein.

Full Text

The Full Text of this article is available as a PDF (186.4 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. Allard J. D., Bertrand K. P. Sequence of a class E tetracycline resistance gene from Escherichia coli and comparison of related tetracycline efflux proteins. J Bacteriol. 1993 Jul;175(14):4554–4560. doi: 10.1128/jb.175.14.4554-4560.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chopra I., Hawkey P. M., Hinton M. Tetracyclines, molecular and clinical aspects. J Antimicrob Chemother. 1992 Mar;29(3):245–277. doi: 10.1093/jac/29.3.245. [DOI] [PubMed] [Google Scholar]
  4. Curiale M. S., Levy S. B. Two complementation groups mediate tetracycline resistance determined by Tn10. J Bacteriol. 1982 Jul;151(1):209–215. doi: 10.1128/jb.151.1.209-215.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Curiale M. S., McMurry L. M., Levy S. B. Intracistronic complementation of the tetracycline resistance membrane protein of Tn10. J Bacteriol. 1984 Jan;157(1):211–217. doi: 10.1128/jb.157.1.211-217.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dosch D. C., Salvacion F. F., Epstein W. Tetracycline resistance element of pBR322 mediates potassium transport. J Bacteriol. 1984 Dec;160(3):1188–1190. doi: 10.1128/jb.160.3.1188-1190.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Hickman R. K., Levy S. B. Evidence that TET protein functions as a multimer in the inner membrane of Escherichia coli. J Bacteriol. 1988 Apr;170(4):1715–1720. doi: 10.1128/jb.170.4.1715-1720.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaneko M., Yamaguchi A., Sawai T. Energetics of tetracycline efflux system encoded by Tn10 in Escherichia coli. FEBS Lett. 1985 Dec 2;193(2):194–198. doi: 10.1016/0014-5793(85)80149-6. [DOI] [PubMed] [Google Scholar]
  10. Marshall B., Morrissey S., Flynn P., Levy S. B. A new tetracycline-resistance determinant, class E, isolated from Enterobacteriaceae. Gene. 1986;50(1-3):111–117. doi: 10.1016/0378-1119(86)90315-x. [DOI] [PubMed] [Google Scholar]
  11. McMurry L. M., Stephan M., Levy S. B. Decreased function of the class B tetracycline efflux protein Tet with mutations at aspartate 15, a putative intramembrane residue. J Bacteriol. 1992 Oct;174(19):6294–6297. doi: 10.1128/jb.174.19.6294-6297.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. McMurry L., Petrucci R. E., Jr, Levy S. B. Active efflux of tetracycline encoded by four genetically different tetracycline resistance determinants in Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3974–3977. doi: 10.1073/pnas.77.7.3974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. McNicholas P., Chopra I., Rothstein D. M. Genetic analysis of the tetA(C) gene on plasmid pBR322. J Bacteriol. 1992 Dec;174(24):7926–7933. doi: 10.1128/jb.174.24.7926-7933.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mendez B., Tachibana C., Levy S. B. Heterogeneity of tetracycline resistance determinants. Plasmid. 1980 Mar;3(2):99–108. doi: 10.1016/0147-619x(80)90101-8. [DOI] [PubMed] [Google Scholar]
  15. Nguyen T. T., Postle K., Bertrand K. P. Sequence homology between the tetracycline-resistance determinants of Tn10 and pBR322. Gene. 1983 Nov;25(1):83–92. doi: 10.1016/0378-1119(83)90170-1. [DOI] [PubMed] [Google Scholar]
  16. Peden K. W. Revised sequence of the tetracycline-resistance gene of pBR322. Gene. 1983 May-Jun;22(2-3):277–280. doi: 10.1016/0378-1119(83)90112-9. [DOI] [PubMed] [Google Scholar]
  17. Rubin R. A., Levy S. B. Interdomain hybrid Tet proteins confer tetracycline resistance only when they are derived from closely related members of the tet gene family. J Bacteriol. 1990 May;172(5):2303–2312. doi: 10.1128/jb.172.5.2303-2312.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rubin R. A., Levy S. B. Tet protein domains interact productively to mediate tetracycline resistance when present on separate polypeptides. J Bacteriol. 1991 Jul;173(14):4503–4509. doi: 10.1128/jb.173.14.4503-4509.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Speer B. S., Shoemaker N. B., Salyers A. A. Bacterial resistance to tetracycline: mechanisms, transfer, and clinical significance. Clin Microbiol Rev. 1992 Oct;5(4):387–399. doi: 10.1128/cmr.5.4.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Waters S. H., Rogowsky P., Grinsted J., Altenbuchner J., Schmitt R. The tetracycline resistance determinants of RP1 and Tn1721: nucleotide sequence analysis. Nucleic Acids Res. 1983 Sep 10;11(17):6089–6105. doi: 10.1093/nar/11.17.6089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Yamaguchi A., Adachi K., Akasaka T., Ono N., Sawai T. Metal-tetracycline/H+ antiporter of Escherichia coli encoded by a transposon Tn10. Histidine 257 plays an essential role in H+ translocation. J Biol Chem. 1991 Apr 5;266(10):6045–6051. [PubMed] [Google Scholar]
  22. Yamaguchi A., Adachi K., Sawai T. Orientation of the carboxyl terminus of the transposon Tn10-encoded tetracycline resistance protein in Escherichia coli. FEBS Lett. 1990 Jun 4;265(1-2):17–19. doi: 10.1016/0014-5793(90)80872-g. [DOI] [PubMed] [Google Scholar]
  23. Yamaguchi A., Akasaka T., Ono N., Someya Y., Nakatani M., Sawai T. Metal-tetracycline/H+ antiporter of Escherichia coli encoded by transposon Tn10. Roles of the aspartyl residues located in the putative transmembrane helices. J Biol Chem. 1992 Apr 15;267(11):7490–7498. [PubMed] [Google Scholar]
  24. Yamaguchi A., Ono N., Akasaka T., Noumi T., Sawai T. Metal-tetracycline/H+ antiporter of Escherichia coli encoded by a transposon, Tn10. The role of the conserved dipeptide, Ser65-Asp66, in tetracycline transport. J Biol Chem. 1990 Sep 15;265(26):15525–15530. [PubMed] [Google Scholar]
  25. Yamaguchi A., Someya Y., Sawai T. Metal-tetracycline/H+ antiporter of Escherichia coli encoded by transposon Tn10. The role of a conserved sequence motif, GXXXXRXGRR, in a putative cytoplasmic loop between helices 2 and 3. J Biol Chem. 1992 Sep 25;267(27):19155–19162. [PubMed] [Google Scholar]
  26. Yamaguchi A., Udagawa T., Sawai T. Transport of divalent cations with tetracycline as mediated by the transposon Tn10-encoded tetracycline resistance protein. J Biol Chem. 1990 Mar 25;265(9):4809–4813. [PubMed] [Google Scholar]
  27. von Heijne G. Membrane protein structure prediction. Hydrophobicity analysis and the positive-inside rule. J Mol Biol. 1992 May 20;225(2):487–494. doi: 10.1016/0022-2836(92)90934-c. [DOI] [PubMed] [Google Scholar]

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

RESOURCES