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. 1993 Jul;37(7):1456–1462. doi: 10.1128/aac.37.7.1456

Characterization of the aac(6')-Ib gene encoding an aminoglycoside 6'-N-acetyltransferase in Pseudomonas aeruginosa BM2656.

M Galimand 1, T Lambert 1, G Gerbaud 1, P Courvalin 1
PMCID: PMC187994  PMID: 8363376

Abstract

Pseudomonas aeruginosa BM2656 was resistant to tobramycin and susceptible to gentamicin and amikacin by disk diffusion testing. This unusual resistance was not transferable by conjugation to Escherichia coli or P. aeruginosa PAO38, and plasmid DNA was not detected in this strain. A 0.9-kb fragment harboring the tobramycin resistance gene was cloned from BM2656 into pUC18, generating pAT129. Analysis for aminoglycoside-modifying activity in extracts of BM2656 and E. coli harboring pAT129 indicated that tobramycin resistance was due to synthesis of an aminoglycoside 6'-N-acetyltransferase type I [AAC(6')-I] enzyme which modified amikacin and tobramycin. Although amikacin was acetylated, the bactericidal synergism of this aminoglycoside with ceftazidime against BM2656 was minimally affected. The sequence of the DNA fragment was determined. It contained an aac (6')-Ib-like gene and was located downstream from a conserved region related to Tn21. The translated sequence of this aac(6')-Ib gene possessed 99.2% identity with the putative products of the aac(6')-Ib gene cassettes from Serratia marcescens and Klebsiella pneumoniae and 69% identity with the putative aacA(6')-II gene product from P. aeruginosa. We conclude that an aac(6')-Ib gene has spread to the chromosome of P. aeruginosa, probably by transposition.

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Selected References

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  1. Bagdasarian M. M., Amann E., Lurz R., Rückert B., Bagdasarian M. Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors. Gene. 1983 Dec;26(2-3):273–282. doi: 10.1016/0378-1119(83)90197-x. [DOI] [PubMed] [Google Scholar]
  2. Bouanchaud D. H., Scavizzi M. R., Chabbert Y. A. Elimination by ethidium bromide of antibiotic resistance in enterobacteria and staphylococci. J Gen Microbiol. 1968 Dec;54(3):417–425. doi: 10.1099/00221287-54-3-417. [DOI] [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. Collis C. M., Hall R. M. Site-specific deletion and rearrangement of integron insert genes catalyzed by the integron DNA integrase. J Bacteriol. 1992 Mar;174(5):1574–1585. doi: 10.1128/jb.174.5.1574-1585.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Courvalin P., Carlier C., Collatz E. Plasmid-mediated resistance to aminocyclitol antibiotics in group D streptococci. J Bacteriol. 1980 Aug;143(2):541–551. doi: 10.1128/jb.143.2.541-551.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Haas D., Holloway B. W. R factor variants with enhanced sex factor activity in Pseudomonas aeruginosa. Mol Gen Genet. 1976 Mar 30;144(3):243–251. doi: 10.1007/BF00341722. [DOI] [PubMed] [Google Scholar]
  7. Haas M. J., Dowding J. E. Aminoglycoside-modifying enzymes. Methods Enzymol. 1975;43:611–628. doi: 10.1016/0076-6879(75)43124-x. [DOI] [PubMed] [Google Scholar]
  8. Hall R. M., Brookes D. E., Stokes H. W. Site-specific insertion of genes into integrons: role of the 59-base element and determination of the recombination cross-over point. Mol Microbiol. 1991 Aug;5(8):1941–1959. doi: 10.1111/j.1365-2958.1991.tb00817.x. [DOI] [PubMed] [Google Scholar]
  9. Jacob A. E., Grinter N. J. Plasmid RP4 as a vector replicor in genetic engineering. Nature. 1975 Jun 5;255(5508):504–506. doi: 10.1038/255504a0. [DOI] [PubMed] [Google Scholar]
  10. Jacoby G. A., Blaser M. J., Santanam P., Hächler H., Kayser F. H., Hare R. S., Miller G. H. Appearance of amikacin and tobramycin resistance due to 4'-aminoglycoside nucleotidyltransferase [ANT(4')-II] in gram-negative pathogens. Antimicrob Agents Chemother. 1990 Dec;34(12):2381–2386. doi: 10.1128/aac.34.12.2381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Labigne-Roussel A., Gerbaud G., Courvalin P. Translocation of sequences encoding antibiotic resistance from the chromosome to a receptor plasmid in Salmonella ordonez. Mol Gen Genet. 1981;182(3):390–408. doi: 10.1007/BF00293927. [DOI] [PubMed] [Google Scholar]
  12. Lambert T., Gerbaud G., Courvalin P. Transferable amikacin resistance in Acinetobacter spp. due to a new type of 3'-aminoglycoside phosphotransferase. Antimicrob Agents Chemother. 1988 Jan;32(1):15–19. doi: 10.1128/aac.32.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mabilat C., Lourençao-Vital J., Goussard S., Courvalin P. A new example of physical linkage between Tn1 and Tn21: the antibiotic multiple-resistance region of plasmid pCFF04 encoding extended-spectrum beta-lactamase TEM-3. Mol Gen Genet. 1992 Oct;235(1):113–121. doi: 10.1007/BF00286188. [DOI] [PubMed] [Google Scholar]
  14. Meyer J. F., Nies B. A., Wiedemann B. Amikacin resistance mediated by multiresistance transposon Tn2424. J Bacteriol. 1983 Aug;155(2):755–760. doi: 10.1128/jb.155.2.755-760.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nobuta K., Tolmasky M. E., Crosa L. M., Crosa J. H. Sequencing and expression of the 6'-N-acetyltransferase gene of transposon Tn1331 from Klebsiella pneumoniae. J Bacteriol. 1988 Aug;170(8):3769–3773. doi: 10.1128/jb.170.8.3769-3773.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Parent R., Roy P. H. The chloramphenicol acetyltransferase gene of Tn2424: a new breed of cat. J Bacteriol. 1992 May;174(9):2891–2897. doi: 10.1128/jb.174.9.2891-2897.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Schmidt F. R., Nücken E. J., Henschke R. B. Structure and function of hot spots providing signals for site-directed specific recombination and gene expression in Tn21 transposons. Mol Microbiol. 1989 Nov;3(11):1545–1555. doi: 10.1111/j.1365-2958.1989.tb00140.x. [DOI] [PubMed] [Google Scholar]
  19. Shaw K. J., Cramer C. A., Rizzo M., Mierzwa R., Gewain K., Miller G. H., Hare R. S. Isolation, characterization, and DNA sequence analysis of an AAC(6')-II gene from Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1989 Dec;33(12):2052–2062. doi: 10.1128/aac.33.12.2052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Staden R. A new computer method for the storage and manipulation of DNA gel reading data. Nucleic Acids Res. 1980 Aug 25;8(16):3673–3694. doi: 10.1093/nar/8.16.3673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stokes H. W., Hall R. M. A novel family of potentially mobile DNA elements encoding site-specific gene-integration functions: integrons. Mol Microbiol. 1989 Dec;3(12):1669–1683. doi: 10.1111/j.1365-2958.1989.tb00153.x. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. Wilbur W. J., Lipman D. J. Rapid similarity searches of nucleic acid and protein data banks. Proc Natl Acad Sci U S A. 1983 Feb;80(3):726–730. doi: 10.1073/pnas.80.3.726. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]

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