The Penicillin Resistance of Enterococcus Faecalis Jh2-2r Results from an Overproduction of the Low-Affinity Penicillin-Binding Protein Pbp4 and Does Not Involve a Psr-Like Gene
[en] A penicillin-resistant mutant, JH2-2r (MIC 75 microg ml(-1)), was isolated from Enterococcus faecalis JH2-2 (MIC 5 microg ml(-1)) by successive passages on plates containing increasing concentrations of benzylpenicillin. A comparison of the penicillin-binding protein (PBP) profiles in the two strains revealed a more intensely labelled PBP4 in JH2-2r. Because the sequences of the JH2-2 and JH2-2r pbp4 genes were strictly identical, even in their promoter regions, this intensive labelling could only be associated with an overproduction of the low-affinity PBP4. No psr gene analogous to that proposed to act as a regulator of PBP5 synthesis in Enterococcus hirae and Enterococcus faecium could be identified in the vicinity of pbp4 in E. faecalis JH2-2 and JH2-2r. However, a psr-like gene distant from pbp4 was identified. The cloning and sequencing of that psr-like gene from both E. faecalis strains indicated that they were identical. It is therefore postulated that the PBP4 overproduction in E. faecalis JH2-2r results from the modification of an as yet unidentified factor.
Disciplines :
Microbiologie
Auteur, co-auteur :
Duez, Colette ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Zorzi, Willy ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques > Histologie humaine
Sapunaric, Frédéric ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Amoroso, Ana Maria ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Thamm, Iris ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Coyette, Jacques ; Université de Liège - ULiège > Services généraux (Faculté des sciences) > Relations académiques et scientifiques (Sciences)
Langue du document :
Anglais
Titre :
The Penicillin Resistance of Enterococcus Faecalis Jh2-2r Results from an Overproduction of the Low-Affinity Penicillin-Binding Protein Pbp4 and Does Not Involve a Psr-Like Gene
Date de publication/diffusion :
septembre 2001
Titre du périodique :
Microbiology
ISSN :
1350-0872
eISSN :
1465-2080
Maison d'édition :
Society for General Microbiology, Reading RG7 1AG, Royaume-Uni
Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J. (1990) Basic local alignment search tool. J Mol Biol 215:403-410.
Andersen P.S., Martinussen J., Hammer K. (1996) Sequence analysis and identification of the pyrKDbF operon from Lactococcus lactis including a novel gene, pyrK involved in pyrimidine biosynthesis. J Bacteriol 178:5005-5012.
Courvalin P. (1994) Transfer of antibiotic resistance genes between Gram-positive and Gram-negative bacteria. Antimicrob Agents Chemother 38:1447-1451.
Coyette J., Ghuysen J.M., Fontana R. (1978) Solubilization and isolation of the membrane-bound DD-carboxypeptidase of Streptococcus faecalis ATCC 9790. Eur J Biochem 88:297-305.
Coyette J., Ghuysen J.M., Fontana R. (1980) The penicillin-binding proteins in Streptococcus faecalis ATCC 9790. Eur J Biochem 110:445-456.
Devereux J., Haeberli P., Smithies O. (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387-395.
El Kharroubi A., Jacques P., Piras G., Van Beeumen J., Coyette J., Ghuysen J.M. (1991) The Enterococcus hirae R40 penicillin-binding protein 5 and the methicillin-resistant Staphylococcus aureus penicillin-binding 2′ are similar. Biochem J 280:463-469.
Fontana R., Cerini R., Longoni P., Grossato A., Canepari P. (1983) Identification of a streptococcal penicillin-binding protein that reacts very slowly with penicillin. J Bacteriol 155:1343-1350.
Fontana R., Grossato A., Rossi L., Cheng Y.R., Satta G. (1985) Transition from resistance to hypersusceptibility to β-lactam antibiotics associated with loss of low affinity PBP in a Streptococcus faecium mutant highly resistant to penicillin. Antimicrob Agents Chemother 26:678-683.
Fontana R., Aldegheri M., Ligozzi M., Lopez H., Sucari A., Satta G. (1994) Overproduction of a low-affinity penicillin-binding protein and high-level ampicillin resistance in Enterococcus faecium. Antimicrob Agents Chemother 38:1980-1983.
Ghuysen J.M., Frère J.M., Leyh-Bouille M., Nguyen-Distèche M., Coyette J. (1986) Active-site serine D-alanyl-D-alanine-cleaving peptidase-catalysed acyl-transfer reactions. Procedures for studying the penicillin-binding proteins of bacterial plasma membranes. Biochem J 235:159-165.
Goffin C., Ghuysen J.M. (1998) Multimodular penicillin-binding proteins: An enigmatic family of orthologs and paralogs. Microbiol Mol Biol Rev 62:1079-1093.
Griffin A.M., Morris V.J., Gasson M.J. (1996) The cpsABCDE genes involved in polysaccharide production in Streptococcus salivarius spp. thermophilus NCBF2393. Gene 183:23-27.
Gutmann L. (1994) Résistance des entérocoques aux bêtalactamines et conséquences sur les synergies. Méd Mal Infect (spécial) 24:165-171.
Hancock L.E., Gilmore M.S. (2000) Pathogenicity of enterococci. Gram-positive Pathogens , Edited by V. A. Fischetti, R. P. Novick, J. J. Ferretti, D. A. Portnoy & J. I. Road. Washington, DC: ASM Press; 251-288.
Innis M.A., Gelfand D.H., Sninsky J.J., White T.J. PCR Protocols. A Guide to Methods and Applications., New York: Academic Press; 1990.
Jett B.D., Huycke M.M., Gilmore M.S. (1994) Virulence of enterococci. Clin Microbiol Rev 7:462-478.
Klare I., Rodloff A.C., Wagner J., Witte W., Hakenbeck R. (1992) Overproduction of a penicillin-binding protein is not the only mechanism of penicillin resistance in Enterococcus faecium. Antimicrob Agents Chemother 36:783-787.
Kunst F., Ogasawara N., Moszer I. (1997) The complete sequence of the Gram-positive bacterium Bacillus subtilis. Nature , & 148 other authors; 390:249-256.
Laskey R.A. (1980) The use of intensifying screens or organic scintillators for visualizing radioactive molecules, resolved by gel electrophoresis. Methods Enzymol 65:363-371.
Lazarevic V., Margot P., Soldo B., Karamata D. (1992) Sequencing and analysis of the Bacillus subtilis lytRABC divergon: A regulatory unit encompassing the structural genes of the N-acetylmuramoyl-L-alanine amidase and its modifier. J Gen Microbiol 138:1949-1961.
Ligozzi M., Pittaluga F., Fontana R. (1993) Identification of a genetic element (psr) which negatively controls expression of Enterococcus hirae penicillin-binding protein 5. J Bacteriol 175:2046-2051.
Ligozzi M., Pittaluga F., Fontana R. (1996) Modification of the penicillin-binding protein 5 associated with high-level ampicillin resistance of Enterococcus faecium. Antimicrob Agents Chemother 40:354-357.
Lillehang D., Nes I.F., Birkeland N.K. (1997) A highly efficient and stable system for site-specific integration of genes and plasmids into the phage φLC3 attachment site (attB) of the Lactococcus lactis chromosome. Gene 188:129-136.
Lindström E.B., Boman H.G., Steele B.B. (1970) Resistance of Escherichia coli to penicillin. VI. Purification and characterization of the chromosomally mediated penicillinase present in ampA-containing strains. J Bacteriol 101:218-231.
Loureiro Dos Santos A.L., Chopin A. (1987) Shotgun cloning in Streptococcus lactis. FEMS Microbiol Lett 42:209-212.
Mainardi J.L., Billot-Klein D., Coutrot A., Legrand R., Schoot B., Gutmann L. (1998) Resistance to cefotaxime and peptidoglycan composition in Enterococcus faecalis are influenced by exogenous sodium chloride. Microbiology 144:2679-2685.
Massidda O., Dardenne O., Whalen M.B., Zorzi W., Coyette J., Shockman G.D., Daneo-Moore L. (1998) The PBP5 synthesis repressor (psr) gene of Enterococcus hirae ATCC 9790 is substantially longer than previously reported. FEMS Microbiol Lett 166:355-360.
Masson J.M., Labia R. (1983) Synthesis of a 125I-radiolabeled penicillin for penicillin-binding protein studies. Anal Biochem 128:164-168.
Mollerach M., Partoune P., Coyette J., Ghuysen J.M. (1996) Importance of the E46-D160 polypeptide segment of the non-penicillin-binding module for the stability of the low-affinity, multimodular class B penicillin-binding protein 5 of Enterococcus hirae. J Bacteriol 178:1774-1775.
Morona J.K., Morona R., Paton J.C. (1997) Characterization of the locus encoding the Streptococcus pneumoniae type 19F capsular polysaccharide biosynthetic pathway. Mol Microbiol 23:751-753.
Murray B.E. (1990) The life and times of the Enterococcus. Clin Microbiol Rev 3:46-65.
Murray T., Popham D.L., Setlow P. (1996) Identification and characterization of pbpC, the gene encoding Bacillus subtilis penicillin-binding protein 3. J Bacteriol 178:6001-6005.
Nakano Y., Yoshida Y., Yamashita Y., Koga T. (1995) Construction of a series of pACYC-derived plasmid vectors. Gene 162:157-158.
Pearson W.R., Lipman D.J. (1988) Improved tools for biological sequence analysis. Proc Natl Acad Sci U S A 85:2444-2448.
Piras G., El Kharroubi A., Van Beeumen J., Coeme E., Coyette J., Ghuysen J.M. (1990) Characterization of an Enterococcus hirae penicillin-binding protein 3 with low penicillin affinity. J Bacteriol 172:6856-6862.
Piras G., Raze D., El Kharroubi A., Hastir D., Englebert S., Coyette J., Ghuysen J.M. (1993) Cloning and sequencing of the low affinity penicillin-binding protein 3′-encoding gene of Enterococcus hirae S185: Modular design and structural organization of the protein. J Bacteriol 175:2844-2852.
Rice L.B., Carias L.L., Hutton-Thomas R., Sifaoui F., Gutmann L., Rudin S.D. (2001) Penicillin-binding protein 5 and expression of ampicillin resistance in Enterococcus faecium. Antimicrob Agents Chemother 45:1480-1486.
Schmit J.L., Leclercq R., Scheimberg A., Laudaner D.I. (1994) Approche épidémiologique et clinique des entérocoques: Résultats d'une enquête. Méd Mal Infect (spécial) 24:141-148.
Signoretto C., Boaretti M., Canepari P. (1994) Cloning, sequencing and expression in Escherichia coli of the low affinity penicillin-binding protein of Enterococcus faecalis. FEMS Microbiol Lett 123:99-106.
Soldo B., Lazarevic V., Mauël C., Karamata D. (1996) Sequence of the 305°-307° region of the Bacillus subtilis chromosome. Microbiology 142:3079-3088.
Song M.D., Wachi M., Doi M., Ishino F., Matsuhashi M. (1987) Evolution of an inducible penicillin-target protein in methicillin-resistant Staphylococcus aureus. FEBS Lett 221:167-171.
Stingele F., Neeser J.R., Mollet B. (1996) Identification and characterization of the eps (exopolysaccharide) gene cluster from Streptococcus thermophilus Sfi6. J Bacteriol 178:1680-1690.
Van Sinderen D., Karsens H., Kok J., Terpstra P., Ruiters M.H., Venema G., Nautra A. (1996) Sequence analysis and molecular characterization of the temperate lactococcal bacteriophage r1t. Mol Microbiol 19:1343-1355.
Williamson R., Le Bouguénec C., Gutmann L., Horaud T. (1985) One or two low-affinity penicillin-binding proteins may be responsible for the range of susceptibility of Enterococcus faecium to benzylpenicillin. J Gen Microbiol 131:1933-1940.
Williamson R., Gutmann L., Horaud T., Delbos F., Acar J.F. (1986) Use of penicillin-binding proteins for identification of enterococci. J Gen Microbiol 132:1929-1937.
Zorzi W., Zhou X.Y., Dardenne O., Lamotte J., Raze D., Pierre J., Gutmann L., Coyette J. (1996) Structure of the low-affinity penicillin-binding protein 5, PBP5fm, in wild-type and highly penicillin-resistant strains of Enterococcus faecium. J Bacteriol 178:4948-4957.
