Abstract
Chromosomal gene aac(6')-Ii of Enterococcus faecium CIP 54-32, encoding a 6'-N-aminoglycoside acetyltransferase was characterized. The gene was identified as a coding sequence of 549 bp corresponding to a protein with a calculated mass of 20,666 Da. Analysis of the sequence of the deduced protein suggested that it was the second member of a subfamily of AAC(6')-I enzymes. Insertional inactivation of aac(6')-Ii led to aminoglycoside susceptibility of CIP 54-32, suggesting that this gene plays a role in resistance to AAC(6')-I substrates. The gene was detected by DNA hybridization in all 26 strains of E. faecium tested but not in 44 other enterococci of 13 species. These data suggest that the aac(6')-Ii gene is species specific and may be used to identify E. faecium.
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Selected References
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- 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]
- Cruz-Rodz A. L., Gilmore M. S. High efficiency introduction of plasmid DNA into glycine treated Enterococcus faecalis by electroporation. Mol Gen Genet. 1990 Oct;224(1):152–154. doi: 10.1007/BF00259462. [DOI] [PubMed] [Google Scholar]
- 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]
- Devriese L. A., Ceyssens K., Rodrigues U. M., Collins M. D. Enterococcus columbae, a species from pigeon intestines. FEMS Microbiol Lett. 1990 Sep 15;59(3):247–251. doi: 10.1016/0378-1097(90)90228-i. [DOI] [PubMed] [Google Scholar]
- Eliopoulos G. M., Wennersten C., Zighelboim-Daum S., Reiszner E., Goldmann D., Moellering R. C., Jr High-level resistance to gentamicin in clinical isolates of Streptococcus (Enterococcus) faecium. Antimicrob Agents Chemother. 1988 Oct;32(10):1528–1532. doi: 10.1128/aac.32.10.1528. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Facklam R. R., Collins M. D. Identification of Enterococcus species isolated from human infections by a conventional test scheme. J Clin Microbiol. 1989 Apr;27(4):731–734. doi: 10.1128/jcm.27.4.731-734.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ferretti J. J., Gilmore K. S., Courvalin P. Nucleotide sequence analysis of the gene specifying the bifunctional 6'-aminoglycoside acetyltransferase 2"-aminoglycoside phosphotransferase enzyme in Streptococcus faecalis and identification and cloning of gene regions specifying the two activities. J Bacteriol. 1986 Aug;167(2):631–638. doi: 10.1128/jb.167.2.631-638.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Galimand M., Lambert T., Gerbaud G., Courvalin P. Characterization of the aac(6')-Ib gene encoding an aminoglycoside 6'-N-acetyltransferase in Pseudomonas aeruginosa BM2656. Antimicrob Agents Chemother. 1993 Jul;37(7):1456–1462. doi: 10.1128/aac.37.7.1456. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Lambert T., Gerbaud G., Trieu-Cuot P., Courvalin P. Structural relationship between the genes encoding 3'-aminoglycoside phosphotransferases in Campylobacter and in gram-positive cocci. Ann Inst Pasteur Microbiol. 1985 Sep-Oct;136B(2):135–150. doi: 10.1016/s0769-2609(85)80040-5. [DOI] [PubMed] [Google Scholar]
- Le Bouguénec C., de Cespédès G., Horaud T. Presence of chromosomal elements resembling the composite structure Tn3701 in streptococci. J Bacteriol. 1990 Feb;172(2):727–734. doi: 10.1128/jb.172.2.727-734.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Moellering R. C., Jr, Korzeniowski O. M., Sande M. A., Wennersten C. B. Species-specific resistance to antimocrobial synergism in Streptococcus faecium and Streptococcus faecalis. J Infect Dis. 1979 Aug;140(2):203–208. doi: 10.1093/infdis/140.2.203. [DOI] [PubMed] [Google Scholar]
- Moran C. P., Jr, Lang N., LeGrice S. F., Lee G., Stephens M., Sonenshein A. L., Pero J., Losick R. Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982;186(3):339–346. doi: 10.1007/BF00729452. [DOI] [PubMed] [Google Scholar]
- Murray B. E. The life and times of the Enterococcus. Clin Microbiol Rev. 1990 Jan;3(1):46–65. doi: 10.1128/cmr.3.1.46. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ounissi H., Derlot E., Carlier C., Courvalin P. Gene homogeneity for aminoglycoside-modifying enzymes in gram-positive cocci. Antimicrob Agents Chemother. 1990 Nov;34(11):2164–2168. doi: 10.1128/aac.34.11.2164. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pearson W. R., Lipman D. J. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. doi: 10.1073/pnas.85.8.2444. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Ruoff K. L., de la Maza L., Murtagh M. J., Spargo J. D., Ferraro M. J. Species identities of enterococci isolated from clinical specimens. J Clin Microbiol. 1990 Mar;28(3):435–437. doi: 10.1128/jcm.28.3.435-437.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Schmidt F. R., Nücken E. J., Henschke R. B. Nucleotide sequence analysis of 2''-aminoglycoside nucleotidyl-transferase ANT(2'') from Tn4000: its relationship with AAD(3'') and impact on Tn21 evolution. Mol Microbiol. 1988 Nov;2(6):709–717. doi: 10.1111/j.1365-2958.1988.tb00081.x. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- 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]
- Terán F. J., Suárez J. E., Mendoza M. C. Cloning, sequencing, and use as a molecular probe of a gene encoding an aminoglycoside 6'-N-acetyltransferase of broad substrate profile. Antimicrob Agents Chemother. 1991 Apr;35(4):714–719. doi: 10.1128/aac.35.4.714. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Trieu-Cuot P., Carlier C., Poyart-Salmeron C., Courvalin P. An integrative vector exploiting the transposition properties of Tn1545 for insertional mutagenesis and cloning of genes from gram-positive bacteria. Gene. 1991 Sep 30;106(1):21–27. doi: 10.1016/0378-1119(91)90561-o. [DOI] [PubMed] [Google Scholar]
- Trieu-Cuot P., Gerbaud G., Lambert T., Courvalin P. In vivo transfer of genetic information between gram-positive and gram-negative bacteria. EMBO J. 1985 Dec 16;4(13A):3583–3587. doi: 10.1002/j.1460-2075.1985.tb04120.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Zubay G. The isolation and properties of CAP, the catabolite gene activator. Methods Enzymol. 1980;65(1):856–877. doi: 10.1016/s0076-6879(80)65079-4. [DOI] [PubMed] [Google Scholar]