Cargando…
Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium
Enterococcus faecium has become a nosocomial pathogen of major importance, causing infections that are difficult to treat owing to its multi-drug resistance. In particular, resistance to the β-lactam antibiotic ampicillin has become ubiquitous among clinical isolates. Mutations in the low-affinity p...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3386183/ https://www.ncbi.nlm.nih.gov/pubmed/22761597 http://dx.doi.org/10.1371/journal.pgen.1002804 |
_version_ | 1782236941695057920 |
---|---|
author | Zhang, Xinglin Paganelli, Fernanda L. Bierschenk, Damien Kuipers, Annemarie Bonten, Marc J. M. Willems, Rob J. L. van Schaik, Willem |
author_facet | Zhang, Xinglin Paganelli, Fernanda L. Bierschenk, Damien Kuipers, Annemarie Bonten, Marc J. M. Willems, Rob J. L. van Schaik, Willem |
author_sort | Zhang, Xinglin |
collection | PubMed |
description | Enterococcus faecium has become a nosocomial pathogen of major importance, causing infections that are difficult to treat owing to its multi-drug resistance. In particular, resistance to the β-lactam antibiotic ampicillin has become ubiquitous among clinical isolates. Mutations in the low-affinity penicillin binding protein PBP5 have previously been shown to be important for ampicillin resistance in E. faecium, but the existence of additional resistance determinants has been suggested. Here, we constructed a high-density transposon mutant library in E. faecium and developed a transposon mutant tracking approach termed Microarray-based Transposon Mapping (M-TraM), leading to the identification of a compendium of E. faecium genes that contribute to ampicillin resistance. These genes are part of the core genome of E. faecium, indicating a high potential for E. faecium to evolve towards β-lactam resistance. To validate the M-TraM results, we adapted a Cre-lox recombination system to construct targeted, markerless mutants in E. faecium. We confirmed the role of four genes in ampicillin resistance by the generation of targeted mutants and further characterized these mutants regarding their resistance to lysozyme. The results revealed that ddcP, a gene predicted to encode a low-molecular-weight penicillin binding protein with D-alanyl-D-alanine carboxypeptidase activity, was essential for high-level ampicillin resistance. Furthermore, deletion of ddcP sensitized E. faecium to lysozyme and abolished membrane-associated D,D-carboxypeptidase activity. This study has led to the development of a broadly applicable platform for functional genomic-based studies in E. faecium, and it provides a new perspective on the genetic basis of ampicillin resistance in this organism. |
format | Online Article Text |
id | pubmed-3386183 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-33861832012-07-03 Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium Zhang, Xinglin Paganelli, Fernanda L. Bierschenk, Damien Kuipers, Annemarie Bonten, Marc J. M. Willems, Rob J. L. van Schaik, Willem PLoS Genet Research Article Enterococcus faecium has become a nosocomial pathogen of major importance, causing infections that are difficult to treat owing to its multi-drug resistance. In particular, resistance to the β-lactam antibiotic ampicillin has become ubiquitous among clinical isolates. Mutations in the low-affinity penicillin binding protein PBP5 have previously been shown to be important for ampicillin resistance in E. faecium, but the existence of additional resistance determinants has been suggested. Here, we constructed a high-density transposon mutant library in E. faecium and developed a transposon mutant tracking approach termed Microarray-based Transposon Mapping (M-TraM), leading to the identification of a compendium of E. faecium genes that contribute to ampicillin resistance. These genes are part of the core genome of E. faecium, indicating a high potential for E. faecium to evolve towards β-lactam resistance. To validate the M-TraM results, we adapted a Cre-lox recombination system to construct targeted, markerless mutants in E. faecium. We confirmed the role of four genes in ampicillin resistance by the generation of targeted mutants and further characterized these mutants regarding their resistance to lysozyme. The results revealed that ddcP, a gene predicted to encode a low-molecular-weight penicillin binding protein with D-alanyl-D-alanine carboxypeptidase activity, was essential for high-level ampicillin resistance. Furthermore, deletion of ddcP sensitized E. faecium to lysozyme and abolished membrane-associated D,D-carboxypeptidase activity. This study has led to the development of a broadly applicable platform for functional genomic-based studies in E. faecium, and it provides a new perspective on the genetic basis of ampicillin resistance in this organism. Public Library of Science 2012-06-28 /pmc/articles/PMC3386183/ /pubmed/22761597 http://dx.doi.org/10.1371/journal.pgen.1002804 Text en Zhang et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zhang, Xinglin Paganelli, Fernanda L. Bierschenk, Damien Kuipers, Annemarie Bonten, Marc J. M. Willems, Rob J. L. van Schaik, Willem Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium |
title | Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium
|
title_full | Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium
|
title_fullStr | Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium
|
title_full_unstemmed | Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium
|
title_short | Genome-Wide Identification of Ampicillin Resistance Determinants in Enterococcus faecium
|
title_sort | genome-wide identification of ampicillin resistance determinants in enterococcus faecium |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3386183/ https://www.ncbi.nlm.nih.gov/pubmed/22761597 http://dx.doi.org/10.1371/journal.pgen.1002804 |
work_keys_str_mv | AT zhangxinglin genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium AT paganellifernandal genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium AT bierschenkdamien genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium AT kuipersannemarie genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium AT bontenmarcjm genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium AT willemsrobjl genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium AT vanschaikwillem genomewideidentificationofampicillinresistancedeterminantsinenterococcusfaecium |