Skip to main content
PMC home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1995 Aug;39(8):1790–1796. doi: 10.1128/aac.39.8.1790

Cloning and characterization of a 3-N-aminoglycoside acetyltransferase gene, aac(3)-Ib, from Pseudomonas aeruginosa.

L R Schwocho 1, C P Schaffner 1, G H Miller 1, R S Hare 1, K J Shaw 1
PMCID: PMC162827  PMID: 7486920

Abstract

A novel gene encoding an aminoglycoside 3-N-acetyltransferase, which confers resistance to gentamicin, astromicin, and sisomicin, was cloned from Pseudomonas aeruginosa Stone 130. Its sequence was determined and found to show considerable similarity to an aac(3)-I gene previously cloned from R plasmids from Enterobacter, Pseudomonas, and Serratia spp. We have designated the genes from the R plasmids and this work aac(3)-Ia and aac(3)-Ib, respectively. The two aac(3)-I genes share 74% nucleotide identity, and their deduced protein products are 88% similar. These data suggest that the genes derive from a common ancestor. Homology between the flanking sequences of both aac(3)-I genes and other resistance determinants known to reside in integron environments was also observed. Intragenic probes specific for either aac(3)-Ia or aac(3)-Ib were used in hybridization studies with a series of gentamicin-, astromicin-, and sisomicin-resistant clinical isolates. Of 59 clinical isolates tested, no isolates hybridized with both probes, 30 (51%) hybridized with the aac(3)-Ia probe, 12 (20%) hybridized with the aac(3)-Ib probe, and 17 (29%) did not hybridize with either probe. These data suggest the existence of at least one other aac(3)-I gene.

Full Text

The Full Text of this article is available as a PDF (436.1 KB).

Selected References

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

  1. Brzezinska M., Benveniste R., Davies J., Daniels P. J., Weinstein J. Gentamicin resistance in strains of Pseudomonas aeruginosa mediated by enzymatic N-acetylation of the deoxystreptamine moiety. Biochemistry. 1972 Feb 29;11(5):761–765. doi: 10.1021/bi00755a013. [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. Chung C. T., Niemela S. L., Miller R. H. One-step preparation of competent Escherichia coli: transformation and storage of bacterial cells in the same solution. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2172–2175. doi: 10.1073/pnas.86.7.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Collis C. M., Grammaticopoulos G., Briton J., Stokes H. W., Hall R. M. Site-specific insertion of gene cassettes into integrons. Mol Microbiol. 1993 Jul;9(1):41–52. doi: 10.1111/j.1365-2958.1993.tb01667.x. [DOI] [PubMed] [Google Scholar]
  5. Collis C. M., Hall R. M. Expression of antibiotic resistance genes in the integrated cassettes of integrons. Antimicrob Agents Chemother. 1995 Jan;39(1):155–162. doi: 10.1128/aac.39.1.155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dornbusch K., Miller G. H., Hare R. S., Shaw K. J. Resistance to aminoglycoside antibiotics in gram-negative bacilli and staphylococci isolated from blood. Report from a European collaborative study. The ESGAR Study Group (European Study Group on Antibiotic Resistance). J Antimicrob Chemother. 1990 Jul;26(1):131–144. doi: 10.1093/jac/26.1.131. [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. 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. Heim U., Tietze E., Weschke W., Tschäpe H., Wobus U. Nucleotide sequence of a plasmid born streptothricin-acetyl-transferase gene (sat-1). Nucleic Acids Res. 1989 Sep 12;17(17):7103–7103. doi: 10.1093/nar/17.17.7103. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Hsiang M. W., White T. J., Davies J. E. NH2-terminal sequence of the aminoglycoside acetyltransferase (3)-I mediated by plasmid RIP 135. FEBS Lett. 1978 Aug 1;92(1):97–99. doi: 10.1016/0014-5793(78)80730-3. [DOI] [PubMed] [Google Scholar]
  13. Jacoby G. A. Properties of R plasmids determining gentamicin resistance by acetylation in Pseudomonas aeruginosa. Antimicrob Agents Chemother. 1974 Sep;6(3):239–252. doi: 10.1128/aac.6.3.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Javier Terán F., Alvarez M., Suárez J. E., Mendoza M. C. Characterization of two aminoglycoside-(3)-N-acetyltransferase genes and assay as epidemiological probes. J Antimicrob Chemother. 1991 Sep;28(3):333–346. doi: 10.1093/jac/28.3.333. [DOI] [PubMed] [Google Scholar]
  15. Lévesque C., Brassard S., Lapointe J., Roy P. H. Diversity and relative strength of tandem promoters for the antibiotic-resistance genes of several integrons. Gene. 1994 May 3;142(1):49–54. doi: 10.1016/0378-1119(94)90353-0. [DOI] [PubMed] [Google Scholar]
  16. López-Cabrera M., Pérez-González J. A., Heinzel P., Piepersberg W., Jiménez A. Isolation and nucleotide sequencing of an aminocyclitol acetyltransferase gene from Streptomyces rimosus forma paromomycinus. J Bacteriol. 1989 Jan;171(1):321–328. doi: 10.1128/jb.171.1.321-328.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mercier J., Lachapelle J., Couture F., Lafond M., Vézina G., Boissinot M., Levesque R. C. Structural and functional characterization of tnpI, a recombinase locus in Tn21 and related beta-lactamase transposons. J Bacteriol. 1990 Jul;172(7):3745–3757. doi: 10.1128/jb.172.7.3745-3757.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Resistance to aminoglycosides in Pseudomonas. Aminoglycoside Resistance Study Groups. Trends Microbiol. 1994 Oct;2(10):347–353. [PubMed] [Google Scholar]
  20. Salauze D., Davies J. Isolation and characterisation of an aminoglycoside phosphotransferase from neomycin-producing Micromonospora chalcea; comparison with that of Streptomyces fradiae and other producers of 4,6-disubstituted 2-deoxystreptamine antibiotics. J Antibiot (Tokyo) 1991 Dec;44(12):1432–1443. doi: 10.7164/antibiotics.44.1432. [DOI] [PubMed] [Google Scholar]
  21. Salauze D., Perez-Gonzalez J. A., Piepersberg W., Davies J. Characterisation of aminoglycoside acetyltransferase-encoding genes of neomycin-producing Micromonospora chalcea and Streptomyces fradiae. Gene. 1991 May 15;101(1):143–148. doi: 10.1016/0378-1119(91)90237-6. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Shaw K. J., Hare R. S., Sabatelli F. J., Rizzo M., Cramer C. A., Naples L., Kocsi S., Munayyer H., Mann P., Miller G. H. Correlation between aminoglycoside resistance profiles and DNA hybridization of clinical isolates. Antimicrob Agents Chemother. 1991 Nov;35(11):2253–2261. doi: 10.1128/aac.35.11.2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shaw K. J., Rather P. N., Hare R. S., Miller G. H. Molecular genetics of aminoglycoside resistance genes and familial relationships of the aminoglycoside-modifying enzymes. Microbiol Rev. 1993 Mar;57(1):138–163. doi: 10.1128/mr.57.1.138-163.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Sundström L., Rådström P., Swedberg G., Sköld O. Site-specific recombination promotes linkage between trimethoprim- and sulfonamide resistance genes. Sequence characterization of dhfrV and sulI and a recombination active locus of Tn21. Mol Gen Genet. 1988 Aug;213(2-3):191–201. doi: 10.1007/BF00339581. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Walker J. E., Gay N. J., Saraste M., Eberle A. N. DNA sequence around the Escherichia coli unc operon. Completion of the sequence of a 17 kilobase segment containing asnA, oriC, unc, glmS and phoS. Biochem J. 1984 Dec 15;224(3):799–815. doi: 10.1042/bj2240799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Weinstein M. J., Drube C. G., Moss E. L., Jr, Waitz J. A. Microbiologic studies related to bacterial resistance to gentamicin. J Infect Dis. 1971 Dec;124 (Suppl):S11–S17. doi: 10.1093/infdis/124.supplement_1.s11. [DOI] [PubMed] [Google Scholar]
  31. Wiedemann B., Meyer J. F., Zühlsdorf M. T. Insertions of resistance genes into Tn21-like transposons. J Antimicrob Chemother. 1986 Oct;18 (Suppl 100):85–92. doi: 10.1093/jac/18.supplement_c.85. [DOI] [PubMed] [Google Scholar]
  32. Wohlleben W., Arnold W., Bissonnette L., Pelletier A., Tanguay A., Roy P. H., Gamboa G. C., Barry G. F., Aubert E., Davies J. On the evolution of Tn21-like multiresistance transposons: sequence analysis of the gene (aacC1) for gentamicin acetyltransferase-3-I(AAC(3)-I), another member of the Tn21-based expression cassette. Mol Gen Genet. 1989 Jun;217(2-3):202–208. doi: 10.1007/BF02464882. [DOI] [PubMed] [Google Scholar]

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

RESOURCES