Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1997 Dec;41(12):2646–2651. doi: 10.1128/aac.41.12.2646

Loss of intrinsic aminoglycoside resistance in Acinetobacter haemolyticus as a result of three distinct types of alterations in the aac(6')-Ig gene, including insertion of IS17.

E Rudant 1, P Courvalin 1, T Lambert 1
PMCID: PMC164184  PMID: 9420034

Abstract

The distribution of the aac(6')-Ig gene, encoding aminoglycoside 6'-N-acetyltransferase-Ig [AAC(6')-Ig], was studied in 96 Acinetobacter haemolyticus strains and 12 proteolytic Acinetobacter strains, including Acinetobacter genomospecies 6, 13, and 14 and 3 unnamed species assigned to this genomic group by DNA-DNA hybridization. This gene was detected by DNA-DNA hybridization in all 96 A. haemolyticus strains and by PCR in 95 strains but was not detected in strains of other species, indicating that it may be used to identify A. haemolyticus. Three A. haemolyticus strains were susceptible to tobramycin and did not produce an aminoglycoside 6'-N-acetylating activity, although they contained aac(6')-Ig-related sequences. An analysis of three susceptible A. haemolyticus strains indicated that aminoglycoside resistance was abolished by the following three distinct mechanisms: (i) a point mutation in aac(6')-Ig that led to a Met56-->Arg substitution, which was shown by analysis of a revertant to be responsible for the loss of resistance; (ii) a polythymine insertion that altered the reading frame; and (iii) insertion of IS17, a new member of the IS903 family. These observations indicated that AAC(6')-Ig is not essential for the viability of A. haemolyticus, although the aac(6')-Ig gene was detected in all members of this species.

Full Text

The Full Text of this article is available as a PDF (242.9 KB).

Selected References

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

  1. Arthur M., Molinas C., Depardieu F., Courvalin P. Characterization of Tn1546, a Tn3-related transposon conferring glycopeptide resistance by synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147. J Bacteriol. 1993 Jan;175(1):117–127. doi: 10.1128/jb.175.1.117-127.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bouvet P. J., Grimont P. A. Identification and biotyping of clinical isolates of Acinetobacter. Ann Inst Pasteur Microbiol. 1987 Sep-Oct;138(5):569–578. doi: 10.1016/0769-2609(87)90042-1. [DOI] [PubMed] [Google Scholar]
  4. Bouvet P. J., Jeanjean S. Delineation of new proteolytic genomic species in the genus Acinetobacter. Res Microbiol. 1989 May-Jun;140(4-5):291–299. doi: 10.1016/0923-2508(89)90021-1. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Costa Y., Galimand M., Leclercq R., Duval J., Courvalin P. Characterization of the chromosomal aac(6')-Ii gene specific for Enterococcus faecium. Antimicrob Agents Chemother. 1993 Sep;37(9):1896–1903. doi: 10.1128/aac.37.9.1896. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Grindley N. D., Joyce C. M. Genetic and DNA sequence analysis of the kanamycin resistance transposon Tn903. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7176–7180. doi: 10.1073/pnas.77.12.7176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Holton T. A., Graham M. W. A simple and efficient method for direct cloning of PCR products using ddT-tailed vectors. Nucleic Acids Res. 1991 Mar 11;19(5):1156–1156. doi: 10.1093/nar/19.5.1156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Juni E. Interspecies transformation of Acinetobacter: genetic evidence for a ubiquitous genus. J Bacteriol. 1972 Nov;112(2):917–931. doi: 10.1128/jb.112.2.917-931.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kholodii G. Y., Gorlenko Z., Lomovskaya O. L., Mindlin S. Z., Yurieva O. V., Nikiforov V. G. Molecular characterization of an aberrant mercury resistance transposable element from an environmental Acinetobacter strain. Plasmid. 1993 Nov;30(3):303–308. doi: 10.1006/plas.1993.1064. [DOI] [PubMed] [Google Scholar]
  12. Lambert T., Gerbaud G., Courvalin P. Characterization of the chromosomal aac(6')-Ij gene of Acinetobacter sp. 13 and the aac(6')-Ih plasmid gene of Acinetobacter baumannii. Antimicrob Agents Chemother. 1994 Sep;38(9):1883–1889. doi: 10.1128/aac.38.9.1883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lambert T., Gerbaud G., Galimand M., Courvalin P. Characterization of Acinetobacter haemolyticus aac(6')-Ig gene encoding an aminoglycoside 6'-N-acetyltransferase which modifies amikacin. Antimicrob Agents Chemother. 1993 Oct;37(10):2093–2100. doi: 10.1128/aac.37.10.2093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Low B. Formation of merodiploids in matings with a class of Rec- recipient strains of Escherichia coli K12. Proc Natl Acad Sci U S A. 1968 May;60(1):160–167. doi: 10.1073/pnas.60.1.160. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Makris J. C., Nordmann P. L., Reznikoff W. S. Mutational analysis of insertion sequence 50 (IS50) and transposon 5 (Tn5) ends. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2224–2228. doi: 10.1073/pnas.85.7.2224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Payie K. G., Rather P. N., Clarke A. J. Contribution of gentamicin 2'-N-acetyltransferase to the O acetylation of peptidoglycan in Providencia stuartii. J Bacteriol. 1995 Aug;177(15):4303–4310. doi: 10.1128/jb.177.15.4303-4310.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ploy M. C., Giamarellou H., Bourlioux P., Courvalin P., Lambert T. Detection of aac(6')-I genes in amikacin-resistant Acinetobacter spp. by PCR. Antimicrob Agents Chemother. 1994 Dec;38(12):2925–2928. doi: 10.1128/aac.38.12.2925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rather P. N., Munayyer H., Mann P. A., Hare R. S., Miller G. H., Shaw K. J. Genetic analysis of bacterial acetyltransferases: identification of amino acids determining the specificities of the aminoglycoside 6'-N-acetyltransferase Ib and IIa proteins. J Bacteriol. 1992 May;174(10):3196–3203. doi: 10.1128/jb.174.10.3196-3203.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Rather P. N., Orosz E., Shaw K. J., Hare R., Miller G. Characterization and transcriptional regulation of the 2'-N-acetyltransferase gene from Providencia stuartii. J Bacteriol. 1993 Oct;175(20):6492–6498. doi: 10.1128/jb.175.20.6492-6498.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Roussel A. F., Chabbert Y. A. Taxonomy and epidemiology of gram-negative bacterial plasmids studied by DNA-DNA filter hybridization in formamide. J Gen Microbiol. 1978 Feb;104(2):269–276. doi: 10.1099/00221287-104-2-269. [DOI] [PubMed] [Google Scholar]
  21. Rudant E., Bourlioux P., Courvalin P., Lambert T. Characterization of the aac(6')-Ik gene of Acinetobacter sp. 6. FEMS Microbiol Lett. 1994 Nov 15;124(1):49–54. doi: 10.1016/0378-1097(94)90331-x. [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., 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]
  24. Shaw K. J., Rather P. N., Sabatelli F. J., Mann P., Munayyer H., Mierzwa R., Petrikkos G. L., Hare R. S., Miller G. H., Bennett P. Characterization of the chromosomal aac(6')-Ic gene from Serratia marcescens. Antimicrob Agents Chemother. 1992 Jul;36(7):1447–1455. doi: 10.1128/aac.36.7.1447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Snelling A. M., Hawkey P. M., Heritage J., Downey P., Bennett P. M., Holmes B. The use of a DNA probe and PCR to examine the distribution of the aac(6')-Ic gene in Serratia marcescens and other gram-negative bacteria. J Antimicrob Chemother. 1993 Jun;31(6):841–854. doi: 10.1093/jac/31.6.841. [DOI] [PubMed] [Google Scholar]
  26. Terai A., Baba K., Shirai H., Yoshida O., Takeda Y., Nishibuchi M. Evidence for insertion sequence-mediated spread of the thermostable direct hemolysin gene among Vibrio species. J Bacteriol. 1991 Aug;173(16):5036–5046. doi: 10.1128/jb.173.16.5036-5046.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Trieu-Cuot P., Courvalin P. Transposition behavior of IS15 and its progenitor IS15-delta: are cointegrates exclusive end products? Plasmid. 1985 Jul;14(1):80–89. doi: 10.1016/0147-619x(85)90034-4. [DOI] [PubMed] [Google Scholar]
  28. Wagner L. A., Weiss R. B., Driscoll R., Dunn D. S., Gesteland R. F. Transcriptional slippage occurs during elongation at runs of adenine or thymine in Escherichia coli. Nucleic Acids Res. 1990 Jun 25;18(12):3529–3535. doi: 10.1093/nar/18.12.3529. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]

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

RESOURCES