Abstract
We report the sequence of a 630-bp fragment of a gene associated with resistance to high levels of vancomycin in a clinical isolate of Enterococcus faecalis which retained susceptibility to teicoplanin. This gene was similar to the recently sequenced vanB and partially homologous with vanA, but it showed less-marked similarity to vanC. A DNA probe, derived from this polymerase chain reaction-amplified gene fragment, hybridized specifically with genomic DNA from Enterococcus faecium and E. faecalis isolates which were vancomycin resistant (MICs ranged from 8 to 512 micrograms/ml) but susceptible to teicoplanin. Curing of vancomycin resistance was associated with loss of DNA hybridization with the gene probe. Transfer of DNA which hybridized with the probe accompanied transfer of vancomycin resistance to a susceptible recipient strain. Neither curing nor transfer of vancomycin resistance was consistently related to loss or acquisition, respectively, of plasmid DNA.
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Selected References
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- Arthur M., Molinas C., Courvalin P. The VanS-VanR two-component regulatory system controls synthesis of depsipeptide peptidoglycan precursors in Enterococcus faecium BM4147. J Bacteriol. 1992 Apr;174(8):2582–2591. doi: 10.1128/jb.174.8.2582-2591.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Arthur M., Molinas C., Dutka-Malen S., Courvalin P. Structural relationship between the vancomycin resistance protein VanH and 2-hydroxycarboxylic acid dehydrogenases. Gene. 1991 Jul 15;103(1):133–134. doi: 10.1016/0378-1119(91)90405-z. [DOI] [PubMed] [Google Scholar]
- 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]
- Brisson-Noël A., Dutka-Malen S., Molinas C., Leclercq R., Courvalin P. Cloning and heterospecific expression of the resistance determinant vanA encoding high-level resistance to glycopeptides in Enterococcus faecium BM4147. Antimicrob Agents Chemother. 1990 May;34(5):924–927. doi: 10.1128/aac.34.5.924. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bugg T. D., Dutka-Malen S., Arthur M., Courvalin P., Walsh C. T. Identification of vancomycin resistance protein VanA as a D-alanine:D-alanine ligase of altered substrate specificity. Biochemistry. 1991 Feb 26;30(8):2017–2021. doi: 10.1021/bi00222a002. [DOI] [PubMed] [Google Scholar]
- Bugg T. D., Wright G. D., Dutka-Malen S., Arthur M., Courvalin P., Walsh C. T. Molecular basis for vancomycin resistance in Enterococcus faecium BM4147: biosynthesis of a depsipeptide peptidoglycan precursor by vancomycin resistance proteins VanH and VanA. Biochemistry. 1991 Oct 29;30(43):10408–10415. doi: 10.1021/bi00107a007. [DOI] [PubMed] [Google Scholar]
- Courvalin P. Resistance of enterococci to glycopeptides. Antimicrob Agents Chemother. 1990 Dec;34(12):2291–2296. doi: 10.1128/aac.34.12.2291. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Daub E., Zawadzke L. E., Botstein D., Walsh C. T. Isolation, cloning, and sequencing of the Salmonella typhimurium ddlA gene with purification and characterization of its product, D-alanine:D-alanine ligase (ADP forming). Biochemistry. 1988 May 17;27(10):3701–3708. doi: 10.1021/bi00410a027. [DOI] [PubMed] [Google Scholar]
- Dutka-Malen S., Leclercq R., Coutant V., Duval J., Courvalin P. Phenotypic and genotypic heterogeneity of glycopeptide resistance determinants in gram-positive bacteria. Antimicrob Agents Chemother. 1990 Oct;34(10):1875–1879. doi: 10.1128/aac.34.10.1875. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dutka-Malen S., Molinas C., Arthur M., Courvalin P. Sequence of the vanC gene of Enterococcus gallinarum BM4174 encoding a D-alanine:D-alanine ligase-related protein necessary for vancomycin resistance. Gene. 1992 Mar 1;112(1):53–58. doi: 10.1016/0378-1119(92)90302-6. [DOI] [PubMed] [Google Scholar]
- Dutka-Malen S., Molinas C., Arthur M., Courvalin P. The VANA glycopeptide resistance protein is related to D-alanyl-D-alanine ligase cell wall biosynthesis enzymes. Mol Gen Genet. 1990 Dec;224(3):364–372. doi: 10.1007/BF00262430. [DOI] [PubMed] [Google Scholar]
- Eliopoulos G. M., Wennersten C., Moellering R. C., Jr Resistance to beta-lactam antibiotics in Streptococcus faecium. Antimicrob Agents Chemother. 1982 Aug;22(2):295–301. doi: 10.1128/aac.22.2.295. [DOI] [PMC free article] [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]
- Evers S., Sahm D. F., Courvalin P. The vanB gene of vancomycin-resistant Enterococcus faecalis V583 is structurally related to genes encoding D-Ala:D-Ala ligases and glycopeptide-resistance proteins VanA and VanC. Gene. 1993 Feb 14;124(1):143–144. doi: 10.1016/0378-1119(93)90779-3. [DOI] [PubMed] [Google Scholar]
- Handwerger S., Pucci M. J., Kolokathis A. Vancomycin resistance is encoded on a pheromone response plasmid in Enterococcus faecium 228. Antimicrob Agents Chemother. 1990 Feb;34(2):358–360. doi: 10.1128/aac.34.2.358. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jacob A. E., Hobbs S. J. Conjugal transfer of plasmid-borne multiple antibiotic resistance in Streptococcus faecalis var. zymogenes. J Bacteriol. 1974 Feb;117(2):360–372. doi: 10.1128/jb.117.2.360-372.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Karanfil L. V., Murphy M., Josephson A., Gaynes R., Mandel L., Hill B. C., Swenson J. M. A cluster of vancomycin-resistant Enterococcus faecium in an intensive care unit. Infect Control Hosp Epidemiol. 1992 Apr;13(4):195–200. doi: 10.1086/646509. [DOI] [PubMed] [Google Scholar]
- Leclercq R., Derlot E., Duval J., Courvalin P. Plasmid-mediated resistance to vancomycin and teicoplanin in Enterococcus faecium. N Engl J Med. 1988 Jul 21;319(3):157–161. doi: 10.1056/NEJM198807213190307. [DOI] [PubMed] [Google Scholar]
- Leclercq R., Dutka-Malen S., Duval J., Courvalin P. Vancomycin resistance gene vanC is specific to Enterococcus gallinarum. Antimicrob Agents Chemother. 1992 Sep;36(9):2005–2008. doi: 10.1128/aac.36.9.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Livornese L. L., Jr, Dias S., Samel C., Romanowski B., Taylor S., May P., Pitsakis P., Woods G., Kaye D., Levison M. E. Hospital-acquired infection with vancomycin-resistant Enterococcus faecium transmitted by electronic thermometers. Ann Intern Med. 1992 Jul 15;117(2):112–116. doi: 10.7326/0003-4819-117-2-112. [DOI] [PubMed] [Google Scholar]
- Nicas T. I., Wu C. Y., Hobbs J. N., Jr, Preston D. A., Allen N. E. Characterization of vancomycin resistance in Enterococcus faecium and Enterococcus faecalis. Antimicrob Agents Chemother. 1989 Jul;33(7):1121–1124. doi: 10.1128/aac.33.7.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Owens D., Stinson J., Collins P., Johnson A., Tomkin G. H. Improvement in the regulation of cellular cholesterologenesis in diabetes: the effect of reduction in serum cholesterol by simvastatin. Diabet Med. 1991 Feb-Mar;8(2):151–156. doi: 10.1111/j.1464-5491.1991.tb01562.x. [DOI] [PubMed] [Google Scholar]
- Robinson A. C., Kenan D. J., Sweeney J., Donachie W. D. Further evidence for overlapping transcriptional units in an Escherichia coli cell envelope-cell division gene cluster: DNA sequence and transcriptional organization of the ddl ftsQ region. J Bacteriol. 1986 Sep;167(3):809–817. doi: 10.1128/jb.167.3.809-817.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rubin L. G., Tucci V., Cercenado E., Eliopoulos G., Isenberg H. D. Vancomycin-resistant Enterococcus faecium in hospitalized children. Infect Control Hosp Epidemiol. 1992 Dec;13(12):700–705. doi: 10.1086/648342. [DOI] [PubMed] [Google Scholar]
- Sahm D. F., Kissinger J., Gilmore M. S., Murray P. R., Mulder R., Solliday J., Clarke B. In vitro susceptibility studies of vancomycin-resistant Enterococcus faecalis. Antimicrob Agents Chemother. 1989 Sep;33(9):1588–1591. doi: 10.1128/aac.33.9.1588. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shlaes D. M., Bouvet A., Devine C., Shlaes J. H., al-Obeid S., Williamson R. Inducible, transferable resistance to vancomycin in Enterococcus faecalis A256. Antimicrob Agents Chemother. 1989 Feb;33(2):198–203. doi: 10.1128/aac.33.2.198. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shlaes D. M., Etter L., Gutmann L. Synergistic killing of vancomycin-resistant enterococci of classes A, B, and C by combinations of vancomycin, penicillin, and gentamicin. Antimicrob Agents Chemother. 1991 Apr;35(4):776–779. doi: 10.1128/aac.35.4.776. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Uttley A. H., George R. C., Naidoo J., Woodford N., Johnson A. P., Collins C. H., Morrison D., Gilfillan A. J., Fitch L. E., Heptonstall J. High-level vancomycin-resistant enterococci causing hospital infections. Epidemiol Infect. 1989 Aug;103(1):173–181. doi: 10.1017/s0950268800030478. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vincent S., Knight R. G., Green M., Sahm D. F., Shlaes D. M. Vancomycin susceptibility and identification of motile enterococci. J Clin Microbiol. 1991 Oct;29(10):2335–2337. doi: 10.1128/jcm.29.10.2335-2337.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williamson R., Al-Obeid S., Shlaes J. H., Goldstein F. W., Shlaes D. M. Inducible resistance to vancomycin in Enterococcus faecium D366. J Infect Dis. 1989 Jun;159(6):1095–1104. doi: 10.1093/infdis/159.6.1095. [DOI] [PubMed] [Google Scholar]
- Zawadzke L. E., Bugg T. D., Walsh C. T. Existence of two D-alanine:D-alanine ligases in Escherichia coli: cloning and sequencing of the ddlA gene and purification and characterization of the DdlA and DdlB enzymes. Biochemistry. 1991 Feb 12;30(6):1673–1682. doi: 10.1021/bi00220a033. [DOI] [PubMed] [Google Scholar]