Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1983 Jan;153(1):116–123. doi: 10.1128/jb.153.1.116-123.1983

Tn1721-encoded tetracycline resistance: mapping of structural and regulatory genes mediating resistance.

J Altenbuchner, K Schmid, R Schmitt
PMCID: PMC217348  PMID: 6294046

Abstract

The genes encoding inducible tetracycline resistance in Tn1721 were located in a 2.1-kilobase portion of the transposon. Using deletions and insertions, we mapped and characterized two tet genes by their mutant phenotypes. Two tetracycline-inducible polypeptides synthesized in minicells were assigned to the tet genes. The polarity of the tet genes was determined by employing a deletion and a gene fusion which altered the carboxy termini of the polypeptides. The gene responsible for resistance (tetA) encompasses 1,250 base pairs and encodes a membrane-bound protein with an apparent molecular weight of 34,000. The second gene (tetR) encompasses at least 650 base pairs and encodes a soluble 26,000-dalton protein, identified by complementation analysis as the repressor. The two adjacent genes have opposite transcriptional polarity, suggesting that the sites controlling their expression are located in the intercistronic region between tetA and tetR.

Full text

PDF
116

Images in this article

120Image
on p.120

Selected References

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

  1. Beck C. F., Mutzel R., Barbé J., Müller W. A multifunctional gene (tetR) controls Tn10-encoded tetracycline resistance. J Bacteriol. 1982 May;150(2):633–642. doi: 10.1128/jb.150.2.633-642.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Boyer H. W., Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. doi: 10.1016/0022-2836(69)90288-5. [DOI] [PubMed] [Google Scholar]
  4. Büchel D. E., Gronenborn B., Müller-Hill B. Sequence of the lactose permease gene. Nature. 1980 Feb 7;283(5747):541–545. doi: 10.1038/283541a0. [DOI] [PubMed] [Google Scholar]
  5. Chopra I., Howe T. G., Linton A. H., Linton K. B., Richmond M. H., Speller D. C. The tetracyclines: prospects at the beginning of the 1980s. J Antimicrob Chemother. 1981 Jul;8(1):5–21. doi: 10.1093/jac/8.1.5. [DOI] [PubMed] [Google Scholar]
  6. Coetzee J. N., Datta N., Hedges R. W. R factors from Proteus rettgeri. J Gen Microbiol. 1972 Oct;72(3):543–552. doi: 10.1099/00221287-72-3-543. [DOI] [PubMed] [Google Scholar]
  7. Coleman D. C., Foster T. J. Analysis of the reduction in expression of tetracycline resistance determined by transposon Tn10 in the multicopy state. Mol Gen Genet. 1981;182(1):171–177. doi: 10.1007/BF00422786. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Dougan G., Saul M., Twigg A., Gill R., Sherratt D. Polypeptides expressed in Escherichia coli K-12 minicells by transposition elements Tn1 and Tn3. J Bacteriol. 1979 Apr;138(1):48–54. doi: 10.1128/jb.138.1.48-54.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dougan G., Sherratt D. The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet. 1977 Mar 7;151(2):151–160. doi: 10.1007/BF00338689. [DOI] [PubMed] [Google Scholar]
  11. Ehring R., Beyreuther K., Wright J. K., Overath P. In vitro and in vivo products of E. coli lactose permease gene are identical. Nature. 1980 Feb 7;283(5747):537–540. doi: 10.1038/283537a0. [DOI] [PubMed] [Google Scholar]
  12. Jorgensen R. A., Reznikoff W. S. Organization of structural and regulatory genes that mediate tetracycline resistance in transposon Tn10. J Bacteriol. 1979 Jun;138(3):705–714. doi: 10.1128/jb.138.3.705-714.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Levy S. B. R factor proteins synthesized in Escherichia coli minicells: incorporation studies with different R factors and detection of deoxyribonucleic acid-binding proteins. J Bacteriol. 1974 Dec;120(3):1451–1463. doi: 10.1128/jb.120.3.1451-1463.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. McKenney K., Shimatake H., Court D., Schmeissner U., Brady C., Rosenberg M. A system to study promoter and terminator signals recognized by Escherichia coli RNA polymerase. Gene Amplif Anal. 1981;2:383–415. [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. Novick R. P., Clowes R. C., Cohen S. N., Curtiss R., 3rd, Datta N., Falkow S. Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev. 1976 Mar;40(1):168–189. doi: 10.1128/br.40.1.168-189.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Reeve J. Use of minicells for bacteriophage-directed polypeptide synthesis. Methods Enzymol. 1979;68:493–503. doi: 10.1016/0076-6879(79)68038-2. [DOI] [PubMed] [Google Scholar]
  19. Schmitt R., Altenbuchner J., Wiebauer K., Arnold W., Pühler A., Schöffl F. Basis of transposition and gene amplification by Tn1721 and related tetracycline-resistance transposons. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):59–65. doi: 10.1101/sqb.1981.045.01.011. [DOI] [PubMed] [Google Scholar]
  20. Schmitt R., Bernhard E., Mattes R. Characterisation of Tn1721, a new transposon containing tetracycline resistance genes capable of amplification. Mol Gen Genet. 1979 Apr 17;172(1):53–65. doi: 10.1007/BF00276215. [DOI] [PubMed] [Google Scholar]
  21. Schöffl F., Arnold W., Pühler A., Altenbuchner J., Schmitt R. The tetracycline resistance transposons Tn1721 and Tn1771 have three 38-base-pair repeats and generate five-base-pair direct repeats. Mol Gen Genet. 1981;181(1):87–94. doi: 10.1007/BF00339010. [DOI] [PubMed] [Google Scholar]
  22. Shaw W. V., Packman L. C., Burleigh B. D., Dell A., Morris H. R., Hartley B. S. Primary structure of a chloramphenicol acetyltransferase specified by R plasmids. Nature. 1979 Dec 20;282(5741):870–872. doi: 10.1038/282870a0. [DOI] [PubMed] [Google Scholar]
  23. Wallace L. J., Ward J. M., Richmond M. H. The location of sequences of TnA required for the establishment of transposition immunity. Mol Gen Genet. 1981;184(1):80–86. doi: 10.1007/BF00271199. [DOI] [PubMed] [Google Scholar]
  24. Ward J. M., Grinsted J. Physical and genetic analysis of the Inc-W group plasmids R388, Sa, and R7K. Plasmid. 1982 May;7(3):239–250. doi: 10.1016/0147-619x(82)90005-1. [DOI] [PubMed] [Google Scholar]
  25. Wiebauer K., Schraml S., Shales S. W., Schmitt R. Tetracycline resistance transposon Tn1721: recA-dependent gene amplification and expression of tetracycline resistance. J Bacteriol. 1981 Sep;147(3):851–859. doi: 10.1128/jb.147.3.851-859.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Wray L. V., Jr, Jorgensen R. A., Reznikoff W. S. Identification of the tetracycline resistance promoter and repressor in transposon Tn10. J Bacteriol. 1981 Aug;147(2):297–304. doi: 10.1128/jb.147.2.297-304.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Yang H. L., Zubay G., Levy S. B. Synthesis of an R plasmid protein associated with tetracycline resistance is negatively regulated. Proc Natl Acad Sci U S A. 1976 May;73(5):1509–1512. doi: 10.1073/pnas.73.5.1509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Zupancic T. J., King S. R., Pogue-Geile K. L., Jaskunas S. R. Identification of a second tetracycline-inducible polypeptide encoded by Tn10. J Bacteriol. 1980 Oct;144(1):346–355. doi: 10.1128/jb.144.1.346-355.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES