Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1989 Dec;33(12):2052–2062. doi: 10.1128/aac.33.12.2052

Isolation, characterization, and DNA sequence analysis of an AAC(6')-II gene from Pseudomonas aeruginosa.

K J Shaw 1, C A Cramer 1, M Rizzo 1, R Mierzwa 1, K Gewain 1, G H Miller 1, R S Hare 1
PMCID: PMC172821  PMID: 2515793

Abstract

The gene encoding a 6'-N-acetyltransferase, AAC(6')-II, was cloned from Pseudomonas aeruginosa plasmid pSCH884. This gene mediates resistance to gentamicin, tobramycin, and netilmicin but not amikacin or isepamicin. The DNA sequence of the gene and flanking regions was determined. The 5'- and 3'-flanking sequences showed near identity to sequences found abutting a variety of different genes encoding resistance determinants. It is likely that the current structure arose by the integration of the 572-base-pair sequence containing the AAC(6')-II gene into a Tn21-related sequence at the recombinational hot spot, AAAGTT. We have compared the sequence of the AAC(6')-II gene to genes of other 6'-N-acetyltransferases. An AAC(6')-Ib protein (encoded by the aacA4 gene; G. Tran Van Nhieu and E. Collatz, J. Bacteriol. 169:5708-5714, 1987) that results in resistance to amikacin but not gentamicin was found to share 82% sequence similarity with the AAC(6')-II protein. We speculate that these two genes arose from a common ancestor and that the processes of selection and dissemination have led to the observed differences in the spectrum of aminoglycoside resistance.

Full text

PDF
2052

Selected References

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

  1. Bolivar F., Backman K. Plasmids of Escherichia coli as cloning vectors. Methods Enzymol. 1979;68:245–267. doi: 10.1016/0076-6879(79)68018-7. [DOI] [PubMed] [Google Scholar]
  2. Cameron F. H., Groot Obbink D. J., Ackerman V. P., Hall R. M. Nucleotide sequence of the AAD(2'') aminoglycoside adenylyltransferase determinant aadB. Evolutionary relationship of this region with those surrounding aadA in R538-1 and dhfrII in R388. Nucleic Acids Res. 1986 Nov 11;14(21):8625–8635. doi: 10.1093/nar/14.21.8625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Champion H. M., Bennett P. M., Lewis D. A., Reeves D. S. Cloning and characterization of an AAC(6') gene from Serratia marcescens. J Antimicrob Chemother. 1988 Nov;22(5):587–596. doi: 10.1093/jac/22.5.587. [DOI] [PubMed] [Google Scholar]
  4. Chinault A. C., Blakesley V. A., Roessler E., Willis D. G., Smith C. A., Cook R. G., Fenwick R. G., Jr Characterization of transferable plasmids from Shigella flexneri 2a that confer resistance to trimethoprim, streptomycin, and sulfonamides. Plasmid. 1986 Mar;15(2):119–131. doi: 10.1016/0147-619x(86)90048-x. [DOI] [PubMed] [Google Scholar]
  5. Courvalin P., Carlier C. Resistance towards aminoglycoside-aminocyclitol antibiotics in bacteria. J Antimicrob Chemother. 1981 Jul;8 (Suppl A):57–69. doi: 10.1093/jac/8.suppl_a.57. [DOI] [PubMed] [Google Scholar]
  6. Dale J. W., Godwin D., Mossakowska D., Stephenson P., Wall S. Sequence of the OXA2 beta-lactamase: comparison with other penicillin-reactive enzymes. FEBS Lett. 1985 Oct 21;191(1):39–44. doi: 10.1016/0014-5793(85)80989-3. [DOI] [PubMed] [Google Scholar]
  7. Fling M. E., Kopf J., Richards C. Nucleotide sequence of the transposon Tn7 gene encoding an aminoglycoside-modifying enzyme, 3"(9)-O-nucleotidyltransferase. Nucleic Acids Res. 1985 Oct 11;13(19):7095–7106. doi: 10.1093/nar/13.19.7095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Guyer M. S. The gamma delta sequence of F is an insertion sequence. J Mol Biol. 1978 Dec 15;126(3):347–365. doi: 10.1016/0022-2836(78)90045-1. [DOI] [PubMed] [Google Scholar]
  9. Hall R. M., Vockler C. The region of the IncN plasmid R46 coding for resistance to beta-lactam antibiotics, streptomycin/spectinomycin and sulphonamides is closely related to antibiotic resistance segments found in IncW plasmids and in Tn21-like transposons. Nucleic Acids Res. 1987 Sep 25;15(18):7491–7501. doi: 10.1093/nar/15.18.7491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hollingshead S., Vapnek D. Nucleotide sequence analysis of a gene encoding a streptomycin/spectinomycin adenylyltransferase. Plasmid. 1985 Jan;13(1):17–30. doi: 10.1016/0147-619x(85)90052-6. [DOI] [PubMed] [Google Scholar]
  11. Hyde D. R., Tu C. P. tnpM: a novel regulatory gene that enhances Tn21 transposition and suppresses cointegrate resolution. Cell. 1985 Sep;42(2):629–638. doi: 10.1016/0092-8674(85)90120-5. [DOI] [PubMed] [Google Scholar]
  12. Kopecko D. J., Brevet J., Cohen S. N. Involvement of multiple translocating DNA segments and recombinational hotspots in the structural evolution of bacterial plasmids. J Mol Biol. 1976 Dec;108(2):333–360. doi: 10.1016/s0022-2836(76)80124-6. [DOI] [PubMed] [Google Scholar]
  13. Lovering A. M., White L. O., Reeves D. S. AAC(1): a new aminoglycoside-acetylating enzyme modifying the Cl aminogroup of apramycin. J Antimicrob Chemother. 1987 Dec;20(6):803–813. doi: 10.1093/jac/20.6.803. [DOI] [PubMed] [Google Scholar]
  14. Lovering A. M., White L. O., Reeves D. S. Identification of aminoglycoside-acetylating enzymes by high-pressure liquid chromatographic determination of their reaction products. Antimicrob Agents Chemother. 1984 Jul;26(1):10–12. doi: 10.1128/aac.26.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McClure W. R. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem. 1985;54:171–204. doi: 10.1146/annurev.bi.54.070185.001131. [DOI] [PubMed] [Google Scholar]
  17. Mercer A. A., Loutit J. S. Transformation and transfection of Pseudomonas aeruginosa: effects of metal ions. J Bacteriol. 1979 Oct;140(1):37–42. doi: 10.1128/jb.140.1.37-42.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meyer J. F., Wiedemann B. Characterization of aminoglycoside 6'-N-acetyltransferases [AAC(6')] from gram-negative bacteria and Streptomyces kanamyceticus. J Antimicrob Chemother. 1985 Mar;15(3):271–282. doi: 10.1093/jac/15.3.271. [DOI] [PubMed] [Google Scholar]
  19. Ouellette M., Bissonnette L., Roy P. H. Precise insertion of antibiotic resistance determinants into Tn21-like transposons: nucleotide sequence of the OXA-1 beta-lactamase gene. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7378–7382. doi: 10.1073/pnas.84.21.7378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pemberton J. M., Holloway B. W. Chromosome mapping in Pseudomonas aeruginosa. Genet Res. 1972 Jun;19(3):251–260. doi: 10.1017/s0016672300014518. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Shaw K. J., Frommer B. R., Anagnost J. A., Narula S., Leibowitz P. J. Regulated secretion of MuGM-CSF in Saccharomyces cerevisiae via GAL1:MF alpha 1 prepro sequences. DNA. 1988 Mar;7(2):117–126. doi: 10.1089/dna.1988.7.117. [DOI] [PubMed] [Google Scholar]
  23. Shimizu K., Kumada T., Hsieh W. C., Chung H. Y., Chong Y., Hare R. S., Miller G. H., Sabatelli F. J., Howard J. Comparison of aminoglycoside resistance patterns in Japan, Formosa, and Korea, Chile, and the United States. Antimicrob Agents Chemother. 1985 Aug;28(2):282–288. doi: 10.1128/aac.28.2.282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Swift G., McCarthy B. J., Heffron F. DNA sequence of a plasmid-encoded dihydrofolate reductase. Mol Gen Genet. 1981;181(4):441–447. doi: 10.1007/BF00428733. [DOI] [PubMed] [Google Scholar]
  25. Tenover F. C., Filpula D., Phillips K. L., Plorde J. J. Cloning and sequencing of a gene encoding an aminoglycoside 6'-N-acetyltransferase from an R factor of Citrobacter diversus. J Bacteriol. 1988 Jan;170(1):471–473. doi: 10.1128/jb.170.1.471-473.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tenover F. C., Phillips K. L., Gilbert T., Lockhart P., O'Hara P. J., Plorde J. J. Development of a DNA probe from the deoxyribonucleotide sequence of a 3-N-aminoglycoside acetyltransferase [AAC(3)-I] resistance gene. Antimicrob Agents Chemother. 1989 Apr;33(4):551–559. doi: 10.1128/aac.33.4.551. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Tran van Nhieu G., Collatz E. Primary structure of an aminoglycoside 6'-N-acetyltransferase AAC(6')-4, fused in vivo with the signal peptide of the Tn3-encoded beta-lactamase. J Bacteriol. 1987 Dec;169(12):5708–5714. doi: 10.1128/jb.169.12.5708-5714.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wang M. D., Liu L., Wang B. M., Berg C. M. Cloning and characterization of the Escherichia coli K-12 alanine-valine transaminase (avtA) gene. J Bacteriol. 1987 Sep;169(9):4228–4234. doi: 10.1128/jb.169.9.4228-4234.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zolg J. W., Hänggi U. J. Characterization of a R plasmid-associated, trimethoprim-resistant dihydrofolate reductase and determination of the nucleotide sequence of the reductase gene. Nucleic Acids Res. 1981 Feb 11;9(3):697–710. doi: 10.1093/nar/9.3.697. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES