Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1994 Mar;38(3):473–475. doi: 10.1128/aac.38.3.473

Alterations in peptidoglycan precursors and vancomycin susceptibility in Tn917 insertion mutants of Enterococcus faecalis 221.

S Handwerger 1
PMCID: PMC284482  PMID: 8203839

Abstract

Derivatives of the highly vancomycin-resistant Enterococcus faecalis strain 221 (MIC, 1,024 micrograms/ml) harboring Tn917 insertions in vanR, vanH, and vanA were compared with the parent strain and the susceptible plasmid-free strain JH2-2 (MIC, 2 micrograms/ml). Cytoplasmic pools of UDP-N-acetyl-muramyl-peptide precursors of strain 221 contained the depsipeptide-terminating precursor as well as elevated levels of both the tripeptide and tetrapeptide precursors. Insertional inactivation of vanR resulted in the loss of carboxypeptidase activity, full susceptibility to vancomycin, and precursor pools similar to those of JH2-2. For the vanA insertional mutant the MBC of vancomycin was fourfold higher than that for JH2-2, and the mutant had increased levels of tripeptide and tetrapeptide precursors compared with those for JH2-2. The vanH insertional mutant showed elevated levels of these precursors, as well as a small amount of depsipeptide, and both the MIC and the MBC of vancomycin were increased compared with those for JH2-2. These findings suggest that DD-carboxypeptidase activity, under the control of vanR, results in increased pools of both tripeptide and tetrapeptide precursors, which may contribute to survival in the presence of vancomycin.

Full text

PDF
473

Selected References

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

  1. Allen N. E., Hobbs J. N., Jr, Richardson J. M., Riggin R. M. Biosynthesis of modified peptidoglycan precursors by vancomycin-resistant Enterococcus faecium. FEMS Microbiol Lett. 1992 Nov 1;77(1-3):109–115. doi: 10.1016/0378-1097(92)90140-j. [DOI] [PubMed] [Google Scholar]
  2. Arthur M., Molinas C., Courvalin P. Sequence of the vanY gene required for production of a vancomycin-inducible D,D-carboxypeptidase in Enterococcus faecium BM4147. Gene. 1992 Oct 12;120(1):111–114. doi: 10.1016/0378-1119(92)90017-j. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. 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]
  5. Billot-Klein D., Gutmann L., Collatz E., van Heijenoort J. Analysis of peptidoglycan precursors in vancomycin-resistant enterococci. Antimicrob Agents Chemother. 1992 Jul;36(7):1487–1490. doi: 10.1128/aac.36.7.1487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Gutmann L., Billot-Klein D., al-Obeid S., Klare I., Francoual S., Collatz E., van Heijenoort J. Inducible carboxypeptidase activity in vancomycin-resistant enterococci. Antimicrob Agents Chemother. 1992 Jan;36(1):77–80. doi: 10.1128/aac.36.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hammes W. P., Kandler O. Biosynthesis of peptidoglycan in Gaffkya homari. The incorporation of peptidoglycan into the cell wall and the direction of transpeptidation. Eur J Biochem. 1976 Nov 1;70(1):97–106. doi: 10.1111/j.1432-1033.1976.tb10960.x. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Handwerger S., Pucci M. J., Volk K. J., Liu J., Lee M. S. The cytoplasmic peptidoglycan precursor of vancomycin-resistant Enterococcus faecalis terminates in lactate. J Bacteriol. 1992 Sep;174(18):5982–5984. doi: 10.1128/jb.174.18.5982-5984.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Handwerger S., Tomasz A. Alterations in kinetic properties of penicillin-binding proteins of penicillin-resistant Streptococcus pneumoniae. Antimicrob Agents Chemother. 1986 Jul;30(1):57–63. doi: 10.1128/aac.30.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Messer J., Reynolds P. E. Modified peptidoglycan precursors produced by glycopeptide-resistant enterococci. FEMS Microbiol Lett. 1992 Jul 1;73(1-2):195–200. doi: 10.1016/0378-1097(92)90608-q. [DOI] [PubMed] [Google Scholar]
  15. Perkins H. R. Specificity of combination between mucopeptide precursors and vancomycin or ristocetin. Biochem J. 1969 Jan;111(2):195–205. doi: 10.1042/bj1110195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Weaver K. E., Clewell D. B. Regulation of the pAD1 sex pheromone response in Enterococcus faecalis: construction and characterization of lacZ transcriptional fusions in a key control region of the plasmid. J Bacteriol. 1988 Sep;170(9):4343–4352. doi: 10.1128/jb.170.9.4343-4352.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wright G. D., Molinas C., Arthur M., Courvalin P., Walsh C. T. Characterization of vanY, a DD-carboxypeptidase from vancomycin-resistant Enterococcus faecium BM4147. Antimicrob Agents Chemother. 1992 Jul;36(7):1514–1518. doi: 10.1128/aac.36.7.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. al-Obeid S., Collatz E., Gutmann L. Mechanism of resistance to vancomycin in Enterococcus faecium D366 and Enterococcus faecalis A256. Antimicrob Agents Chemother. 1990 Feb;34(2):252–256. doi: 10.1128/aac.34.2.252. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES