This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Arbeloa, A.
Right arrow Articles by Arthur, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Arbeloa, A.
Right arrow Articles by Arthur, M.

Next Article 

Journal of Bacteriology, March 2004, p. 1221-1228, Vol. 186, No. 5
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.5.1221-1228.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Role of Class A Penicillin-Binding Proteins in PBP5-Mediated ß-Lactam Resistance in Enterococcus faecalis

Ana Arbeloa,1 Heidi Segal,1,{dagger} Jean-Emmanuel Hugonnet,1 Nathalie Josseaume,1 Lionnel Dubost,2 Jean-Paul Brouard,2 Laurent Gutmann,1 Dominique Mengin-Lecreulx,3 and Michel Arthur1*

INSERM E0004-LRMA, Université Paris VI, 75270 Paris,1 Département Régulations, Développement et Diversité Moléculaire, Museum National d'Histoire Naturelle, USM0502-CNRS UMR8041, 75005 Paris,2 Institut de Biochimie et de Biophysique Moléculaire et Cellulaire, UMR 8619 CNRS, Université Paris-Sud, 91405 Orsay, France3

Received 21 July 2003/ Accepted 21 November 2003

Peptidoglycan polymerization complexes contain multimodular penicillin-binding proteins (PBP) of classes A and B that associate a conserved C-terminal transpeptidase module to an N-terminal glycosyltransferase or morphogenesis module, respectively. In Enterococcus faecalis, class B PBP5 mediates intrinsic resistance to the cephalosporin class of ß-lactam antibiotics, such as ceftriaxone. To identify the glycosyltransferase partner(s) of PBP5, combinations of deletions were introduced in all three class A PBP genes of E. faecalis JH2-2 (ponA, pbpF, and pbpZ). Among mutants with single or double deletions, only JH2-2 {Delta}ponA {Delta}pbpF was susceptible to ceftriaxone. Ceftriaxone resistance was restored by heterologous expression of pbpF from Enterococcus faecium but not by mgt encoding the monofunctional glycosyltransferase of Staphylococcus aureus. Thus, PBP5 partners essential for peptidoglycan polymerization in the presence of ß-lactams formed a subset of the class A PBPs of E. faecalis, and heterospecific complementation was observed with an ortholog from E. faecium. Site-directed mutagenesis of pbpF confirmed that the catalytic serine residue of the transpeptidase module was not required for resistance. None of the three class A PBP genes was essential for viability, although deletion of the three genes led to an increase in the generation time and to a decrease in peptidoglycan cross-linking. As the E. faecalis chromosome does not contain any additional glycosyltransferase-related genes, these observations indicate that glycan chain polymerization in the triple mutant is performed by a novel type of glycosyltransferase. The latter enzyme was not inhibited by moenomycin, since deletion of the three class A PBP genes led to high-level resistance to this glycosyltransferase inhibitor.

* Corresponding author. Mailing address: LRMA, Université Paris VI, 15 rue de l'Ecole de Médecine, 75270 Paris Cedex 06, France. Phone: 33 (0)1 43 25 00 33. Fax: 33 (0)1 43 25 68 12. E-mail: michel.arthur{at}bhdc.jussieu.fr.

{dagger} Present address: Department of Medical Microbiology, Medical School, University of Cape Town, 7925 Cape Town, South Africa.

Journal of Bacteriology, March 2004, p. 1221-1228, Vol. 186, No. 5
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.5.1221-1228.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Vasudevan, P., McElligott, J., Attkisson, C., Betteken, M., Popham, D. L. (2009). Homologues of the Bacillus subtilis SpoVB Protein Are Involved in Cell Wall Metabolism. J. Bacteriol. 191: 6012-6019 [Abstract] [Full Text]  
  • Rice, L. B., Carias, L. L., Rudin, S., Hutton, R., Marshall, S., Hassan, M., Josseaume, N., Dubost, L., Marie, A., Arthur, M. (2009). Role of Class A Penicillin-Binding Proteins in the Expression of {beta}-Lactam Resistance in Enterococcus faecium. J. Bacteriol. 191: 3649-3656 [Abstract] [Full Text]  
  • van Heijenoort, J. (2007). Lipid Intermediates in the Biosynthesis of Bacterial Peptidoglycan. Microbiol. Mol. Biol. Rev. 71: 620-635 [Abstract] [Full Text]  
  • Terrak, M., Nguyen-Disteche, M. (2006). Kinetic Characterization of the Monofunctional Glycosyltransferase from Staphylococcus aureus.. J. Bacteriol. 188: 2528-2532 [Abstract] [Full Text]  
  • Zawadzka-Skomial, J., Markiewicz, Z., Nguyen-Disteche, M., Devreese, B., Frere, J.-M., Terrak, M. (2006). Characterization of the Bifunctional Glycosyltransferase/Acyltransferase Penicillin-Binding Protein 4 of Listeria monocytogenes.. J. Bacteriol. 188: 1875-1881 [Abstract] [Full Text]  
  • Nallapareddy, S. R., Singh, K. V., Murray, B. E. (2006). Construction of Improved Temperature-Sensitive and Mobilizable Vectors and Their Use for Constructing Mutations in the Adhesin-Encoding acm Gene of Poorly Transformable Clinical Enterococcus faecium Strains. Appl. Environ. Microbiol. 72: 334-345 [Abstract] [Full Text]  
  • Kristich, C. J., Manias, D. A., Dunny, G. M. (2005). Development of a Method for Markerless Genetic Exchange in Enterococcus faecalis and Its Use in Construction of a srtA Mutant. Appl. Environ. Microbiol. 71: 5837-5849 [Abstract] [Full Text]  
  • Severin, A., Wu, S. W., Tabei, K., Tomasz, A. (2005). High-Level {beta}-Lactam Resistance and Cell Wall Synthesis Catalyzed by the mecA Homologue of Staphylococcus sciuri Introduced into Staphylococcus aureus. J. Bacteriol. 187: 6651-6658 [Abstract] [Full Text]  
  • Arbeloa, A., Hugonnet, J.-E., Sentilhes, A.-C., Josseaume, N., Dubost, L., Monsempes, C., Blanot, D., Brouard, J.-P., Arthur, M. (2004). Synthesis of Mosaic Peptidoglycan Cross-bridges by Hybrid Peptidoglycan Assembly Pathways in Gram-positive Bacteria. J. Biol. Chem. 279: 41546-41556 [Abstract] [Full Text]  
  • Duez, C., Hallut, S., Rhazi, N., Hubert, S., Amoroso, A., Bouillenne, F., Piette, A., Coyette, J. (2004). The ponA Gene of Enterococcus faecalis JH2-2 Codes for a Low-Affinity Class A Penicillin-Binding Protein. J. Bacteriol. 186: 4412-4416 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»