Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Jan;179(1):97–106. doi: 10.1128/jb.179.1.97-106.1997

The VanS sensor negatively controls VanR-mediated transcriptional activation of glycopeptide resistance genes of Tn1546 and related elements in the absence of induction.

M Arthur 1, F Depardieu 1, G Gerbaud 1, M Galimand 1, R Leclercq 1, P Courvalin 1
PMCID: PMC178666  PMID: 8981985

Abstract

Transposon Tn1546 from Enterococcus faecium BM4147 encodes a histidine protein kinase (VanS) and a response regulator (VanR) that regulate transcription of the vanHAX operon encoding a dehydrogenase (VanH), a ligase (VanA), and a D,D-dipeptidase (VanX). These last three enzymes confer resistance to glycopeptide antibiotics by production of peptidoglycan precursors ending in the depsipeptide D-alanyl-D-lactate. Transcription of vanS and the role of VanS in the regulation of the vanHAX operon were analyzed by inserting a cat reporter gene into vanS. Transcription of cat and vanX was inducible by glycopeptides in partial diploids harboring vanS and vanS(omega)cat but was constitutive in strains containing only vanS(omega)cat. Promoters P(R) and P(H), located upstream from vanR and vanH, respectively, were cloned into a promoter probing vector to study transactivation by chromosomally encoded VanR and VanS. The promoters were inactive in the absence of vanR and vanS, inducible by glycopeptides in the presence of both genes, and constitutively activated by VanR in the absence of VanS. Thus, induction of the vanHAX operon involves an amplification loop resulting from binding of phospho-VanR to the P(R) promoter and increased transcription of the vanR and vanS genes. Full activation of P(R) and P(H) by VanR was observed in the absence of VanS, indicating that the sensor negatively controls VanR in the absence of glycopeptides, presumably by dephosphorylation. Activation of the VanR response regulator in the absence of VanS may involve autophosphorylation of VanR with acetyl phosphate or phosphorylation by a heterologous histidine protein kinase.

Full Text

The Full Text of this article is available as a PDF (359.0 KB).

Selected References

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

  1. Arthur M., Courvalin P. Genetics and mechanisms of glycopeptide resistance in enterococci. Antimicrob Agents Chemother. 1993 Aug;37(8):1563–1571. doi: 10.1128/aac.37.8.1563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Arthur M., Depardieu F., Molinas C., Reynolds P., Courvalin P. The vanZ gene of Tn1546 from Enterococcus faecium BM4147 confers resistance to teicoplanin. Gene. 1995 Feb 27;154(1):87–92. doi: 10.1016/0378-1119(94)00851-i. [DOI] [PubMed] [Google Scholar]
  3. Arthur M., Depardieu F., Reynolds P., Courvalin P. Quantitative analysis of the metabolism of soluble cytoplasmic peptidoglycan precursors of glycopeptide-resistant enterococci. Mol Microbiol. 1996 Jul;21(1):33–44. doi: 10.1046/j.1365-2958.1996.00617.x. [DOI] [PubMed] [Google Scholar]
  4. Arthur M., Depardieu F., Snaith H. A., Reynolds P. E., Courvalin P. Contribution of VanY D,D-carboxypeptidase to glycopeptide resistance in Enterococcus faecalis by hydrolysis of peptidoglycan precursors. Antimicrob Agents Chemother. 1994 Sep;38(9):1899–1903. doi: 10.1128/aac.38.9.1899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Brosius J. Plasmid vectors for the selection of promoters. Gene. 1984 Feb;27(2):151–160. doi: 10.1016/0378-1119(84)90136-7. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Fisher S. L., Jiang W., Wanner B. L., Walsh C. T. Cross-talk between the histidine protein kinase VanS and the response regulator PhoB. Characterization and identification of a VanS domain that inhibits activation of PhoB. J Biol Chem. 1995 Sep 29;270(39):23143–23149. doi: 10.1074/jbc.270.39.23143. [DOI] [PubMed] [Google Scholar]
  11. Handwerger S., Discotto L., Thanassi J., Pucci M. J. Insertional inactivation of a gene which controls expression of vancomycin resistance on plasmid pHKK100. FEMS Microbiol Lett. 1992 Apr 1;71(1):11–14. doi: 10.1016/0378-1097(92)90533-t. [DOI] [PubMed] [Google Scholar]
  12. Handwerger S., Skoble J., Discotto L. F., Pucci M. J. Heterogeneity of the vanA gene cluster in clinical isolates of enterococci from the northeastern United States. Antimicrob Agents Chemother. 1995 Feb;39(2):362–368. doi: 10.1128/aac.39.2.362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Handwerger S., Skoble J. Identification of chromosomal mobile element conferring high-level vancomycin resistance in Enterococcus faecium. Antimicrob Agents Chemother. 1995 Nov;39(11):2446–2453. doi: 10.1128/aac.39.11.2446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Holman T. R., Wu Z., Wanner B. L., Walsh C. T. Identification of the DNA-binding site for the phosphorylated VanR protein required for vancomycin resistance in Enterococcus faecium. Biochemistry. 1994 Apr 19;33(15):4625–4631. doi: 10.1021/bi00181a024. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Kay R., McPherson J. Hybrid pUC vectors for addition of new restriction enzyme sites to the ends of DNA fragments. Nucleic Acids Res. 1987 Mar 25;15(6):2778–2778. doi: 10.1093/nar/15.6.2778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Leclercq R., Derlot E., Weber M., Duval J., Courvalin P. Transferable vancomycin and teicoplanin resistance in Enterococcus faecium. Antimicrob Agents Chemother. 1989 Jan;33(1):10–15. doi: 10.1128/aac.33.1.10. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miranda A. G., Singh K. V., Murray B. E. DNA fingerprinting of Enterococcus faecium by pulsed-field gel electrophoresis may be a useful epidemiologic tool. J Clin Microbiol. 1991 Dec;29(12):2752–2757. doi: 10.1128/jcm.29.12.2752-2757.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reynolds P. E., Depardieu F., Dutka-Malen S., Arthur M., Courvalin P. Glycopeptide resistance mediated by enterococcal transposon Tn1546 requires production of VanX for hydrolysis of D-alanyl-D-alanine. Mol Microbiol. 1994 Sep;13(6):1065–1070. doi: 10.1111/j.1365-2958.1994.tb00497.x. [DOI] [PubMed] [Google Scholar]
  21. Reynolds P. E. Structure, biochemistry and mechanism of action of glycopeptide antibiotics. Eur J Clin Microbiol Infect Dis. 1989 Nov;8(11):943–950. doi: 10.1007/BF01967563. [DOI] [PubMed] [Google Scholar]
  22. Stock J. B., Ninfa A. J., Stock A. M. Protein phosphorylation and regulation of adaptive responses in bacteria. Microbiol Rev. 1989 Dec;53(4):450–490. doi: 10.1128/mr.53.4.450-490.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Walsh C. T., Fisher S. L., Park I. S., Prahalad M., Wu Z. Bacterial resistance to vancomycin: five genes and one missing hydrogen bond tell the story. Chem Biol. 1996 Jan;3(1):21–28. doi: 10.1016/s1074-5521(96)90079-4. [DOI] [PubMed] [Google Scholar]
  25. Wright G. D., Holman T. R., Walsh C. T. Purification and characterization of VanR and the cytosolic domain of VanS: a two-component regulatory system required for vancomycin resistance in Enterococcus faecium BM4147. Biochemistry. 1993 May 18;32(19):5057–5063. doi: 10.1021/bi00070a013. [DOI] [PubMed] [Google Scholar]
  26. Wu Z., Wright G. D., Walsh C. T. Overexpression, purification, and characterization of VanX, a D-, D-dipeptidase which is essential for vancomycin resistance in Enterococcus faecium BM4147. Biochemistry. 1995 Feb 28;34(8):2455–2463. doi: 10.1021/bi00008a008. [DOI] [PubMed] [Google Scholar]
  27. 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 Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES