Undecaprenyl pyrophosphate phosphatase confers low-level resistance to bacitracin in Enterococcus faecalis

A. Shaaly, F. Kalamorz, S. Gebhard, G. M. Cook

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Objectives
Undecaprenyl pyrophosphate phosphatases (UppPs) have been implicated in bacitracin resistance in some bacterial genera and the aim of this study was to determine the role of UppPs in mediating low-level bacitracin resistance in Enterococcus faecalis.
Methods
The uppP gene was identified in the genomes of laboratory (JH2-2) and clinical (V583) strains of E. faecalis. Gene fusions (uppP-lacZ) and 5′-RACE were used to study uppP expression. The uppP gene in both strains was inactivated and mutants were studied for antimicrobial susceptibility and their susceptibilities to various stress agents.
Results
The UppP protein from E. faecalis showed high sequence identity to the Escherichia coli BacA-type UppP and was predicted to be a hydrophobic protein with eight transmembrane helices. The expression of uppP-lacZ was constitutive and not affected by bacitracin or cell wall-active antimicrobials. E. faecalis uppP mutants showed no significant changes in growth rate, colony morphology and biofilm formation. The uppP mutants exhibited increased susceptibility to bacitracin (MICs = 3–6 mg/L) relative to the isogenic wild-type (MICs = 32–48 mg/L). When uppP was expressed in a wild-type background, the MIC of bacitracin increased to 128–≥256 mg/L. The MICs of cefoxitin, teicoplanin, vancomycin, gentamicin, enrofloxacin and D-cycloserine were unaltered in the uppP mutant relative to the wild-type, as were susceptibilities to other stress agents (glycine, lysozyme, NaCl, SDS, low and high pH, oxidative stress and ethanol).
Conclusions
The results demonstrate that low-level bacitracin resistance in E. faecalis is mediated by a BacA-type UppP.
Original languageEnglish
Pages (from-to)1583-1593
JournalJournal of Antimicrobial Chemotherapy
Volume68
Issue number7
DOIs
Publication statusPublished - 1 Jul 2013

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Bacitracin
Enterococcus faecalis
Glycine Agents
Cycloserine
Teicoplanin
Cefoxitin
Gene Fusion
Vancomycin
Biofilms
Muramidase
Gentamicins
Cell Wall
Genes
undecaprenyl pyrophosphate phosphatase
Proteins
Oxidative Stress
Ethanol
Genome
Escherichia coli
Growth

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Undecaprenyl pyrophosphate phosphatase confers low-level resistance to bacitracin in Enterococcus faecalis. / Shaaly, A.; Kalamorz, F.; Gebhard, S.; Cook, G. M.

In: Journal of Antimicrobial Chemotherapy, Vol. 68, No. 7, 01.07.2013, p. 1583-1593.

Research output: Contribution to journalArticle

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title = "Undecaprenyl pyrophosphate phosphatase confers low-level resistance to bacitracin in Enterococcus faecalis",
abstract = "ObjectivesUndecaprenyl pyrophosphate phosphatases (UppPs) have been implicated in bacitracin resistance in some bacterial genera and the aim of this study was to determine the role of UppPs in mediating low-level bacitracin resistance in Enterococcus faecalis.MethodsThe uppP gene was identified in the genomes of laboratory (JH2-2) and clinical (V583) strains of E. faecalis. Gene fusions (uppP-lacZ) and 5′-RACE were used to study uppP expression. The uppP gene in both strains was inactivated and mutants were studied for antimicrobial susceptibility and their susceptibilities to various stress agents.ResultsThe UppP protein from E. faecalis showed high sequence identity to the Escherichia coli BacA-type UppP and was predicted to be a hydrophobic protein with eight transmembrane helices. The expression of uppP-lacZ was constitutive and not affected by bacitracin or cell wall-active antimicrobials. E. faecalis uppP mutants showed no significant changes in growth rate, colony morphology and biofilm formation. The uppP mutants exhibited increased susceptibility to bacitracin (MICs = 3–6 mg/L) relative to the isogenic wild-type (MICs = 32–48 mg/L). When uppP was expressed in a wild-type background, the MIC of bacitracin increased to 128–≥256 mg/L. The MICs of cefoxitin, teicoplanin, vancomycin, gentamicin, enrofloxacin and D-cycloserine were unaltered in the uppP mutant relative to the wild-type, as were susceptibilities to other stress agents (glycine, lysozyme, NaCl, SDS, low and high pH, oxidative stress and ethanol).ConclusionsThe results demonstrate that low-level bacitracin resistance in E. faecalis is mediated by a BacA-type UppP.",
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T1 - Undecaprenyl pyrophosphate phosphatase confers low-level resistance to bacitracin in Enterococcus faecalis

AU - Shaaly, A.

AU - Kalamorz, F.

AU - Gebhard, S.

AU - Cook, G. M.

PY - 2013/7/1

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N2 - ObjectivesUndecaprenyl pyrophosphate phosphatases (UppPs) have been implicated in bacitracin resistance in some bacterial genera and the aim of this study was to determine the role of UppPs in mediating low-level bacitracin resistance in Enterococcus faecalis.MethodsThe uppP gene was identified in the genomes of laboratory (JH2-2) and clinical (V583) strains of E. faecalis. Gene fusions (uppP-lacZ) and 5′-RACE were used to study uppP expression. The uppP gene in both strains was inactivated and mutants were studied for antimicrobial susceptibility and their susceptibilities to various stress agents.ResultsThe UppP protein from E. faecalis showed high sequence identity to the Escherichia coli BacA-type UppP and was predicted to be a hydrophobic protein with eight transmembrane helices. The expression of uppP-lacZ was constitutive and not affected by bacitracin or cell wall-active antimicrobials. E. faecalis uppP mutants showed no significant changes in growth rate, colony morphology and biofilm formation. The uppP mutants exhibited increased susceptibility to bacitracin (MICs = 3–6 mg/L) relative to the isogenic wild-type (MICs = 32–48 mg/L). When uppP was expressed in a wild-type background, the MIC of bacitracin increased to 128–≥256 mg/L. The MICs of cefoxitin, teicoplanin, vancomycin, gentamicin, enrofloxacin and D-cycloserine were unaltered in the uppP mutant relative to the wild-type, as were susceptibilities to other stress agents (glycine, lysozyme, NaCl, SDS, low and high pH, oxidative stress and ethanol).ConclusionsThe results demonstrate that low-level bacitracin resistance in E. faecalis is mediated by a BacA-type UppP.

AB - ObjectivesUndecaprenyl pyrophosphate phosphatases (UppPs) have been implicated in bacitracin resistance in some bacterial genera and the aim of this study was to determine the role of UppPs in mediating low-level bacitracin resistance in Enterococcus faecalis.MethodsThe uppP gene was identified in the genomes of laboratory (JH2-2) and clinical (V583) strains of E. faecalis. Gene fusions (uppP-lacZ) and 5′-RACE were used to study uppP expression. The uppP gene in both strains was inactivated and mutants were studied for antimicrobial susceptibility and their susceptibilities to various stress agents.ResultsThe UppP protein from E. faecalis showed high sequence identity to the Escherichia coli BacA-type UppP and was predicted to be a hydrophobic protein with eight transmembrane helices. The expression of uppP-lacZ was constitutive and not affected by bacitracin or cell wall-active antimicrobials. E. faecalis uppP mutants showed no significant changes in growth rate, colony morphology and biofilm formation. The uppP mutants exhibited increased susceptibility to bacitracin (MICs = 3–6 mg/L) relative to the isogenic wild-type (MICs = 32–48 mg/L). When uppP was expressed in a wild-type background, the MIC of bacitracin increased to 128–≥256 mg/L. The MICs of cefoxitin, teicoplanin, vancomycin, gentamicin, enrofloxacin and D-cycloserine were unaltered in the uppP mutant relative to the wild-type, as were susceptibilities to other stress agents (glycine, lysozyme, NaCl, SDS, low and high pH, oxidative stress and ethanol).ConclusionsThe results demonstrate that low-level bacitracin resistance in E. faecalis is mediated by a BacA-type UppP.

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UR - http://dx.doi.org/10.1093/jac/dkt048

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JO - Journal of Antimicrobial Chemotherapy

JF - Journal of Antimicrobial Chemotherapy

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