title: Sequence of the vanB and ddl genes encoding D-alanine:D-lactate and D-alanine:D-alanine ligases in vancomycin-resistant Enterococcus faecalis V583.
abstract: A pair of degenerate oligodeoxyribonucleotides was used to amplify, by the polymerase chain reaction (PCR), DNA fragments internal to genes encoding D-Ala:D-Ala ligase-related proteins of vancomycin-resistant (VmR) Enterococcus faecalis V583. Cloning and nucleotide sequencing of the PCR products indicated that fragments of two genes, designated vanB and ddl, were co-amplified. The vanB gene was previously shown to be present in Enterococcus strains expressing VanB-type VmR [Quintiliani Jr. et al., J. Infect. Dis. 8 (1993) 943-950]. The ddl gene was detected by Southern hybridization in all VmR and VmS strains of En. faecalis, but not in representatives of 17 other species of Enterococcus. The vanB and ddl genes were cloned in bacteriophage lambda and sequenced. There was extensive similarity (76% amino-acid identity) between the product of vanB and the VmR protein, VanA. The product of ddl, the D-Ala:D-Ala ligases, DdlA and DdlB, of Escherichia coli and the resistance proteins, VanA and VanB, were more distantly related (32-40% aa identity). After induction of VmR, En. faecalis V583 synthesized the cell wall precursor, UDP-N-acetylmuramyl-tetrapeptide-D-lactate, indicating that the mechanism of glycopeptide resistance in strains with the VanA and VanB phenotype is similar.
authors.affiliation: Unité des Agents Antibactériens, Centre National de la Recherche Scientifique EP J0058, Institut Pasteur, Paris, France.
authors.forename: S
authors.initials: S
authors.lastname: Evers
authors.forename: P E
authors.initials: PE
authors.lastname: Reynolds
authors.forename: P
authors.initials: P
authors.lastname: Courvalin
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics*; D001483:Base Sequence; D004269:DNA, Bacterial; D004352:Drug Resistance, Microbial / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics*; D005784:Gene Amplification; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D010453:Peptide Synthases / Q000235:genetics*; D010457:Peptidoglycan / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014640:Vancomycin / Q000494:pharmacology*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D010457:Peptidoglycan; D011498:Protein Precursors; C079913:VanB protein, Enterococcus; D014640:Vancomycin; D010453:Peptide Synthases; C019638:D-alanylalanine synthetase