Diversity of TEM mutants in Proteus mirabilis.
In a survey of resistance to amoxicillin among clinical isolates of Proteus mirabilis, 10 TEM-type beta-lactamases were characterized: (i) the well-known penicillinases TEM-1 and TEM-2, the extended-spectrum beta-lactamases (ESBLs) TEM-3 and TEM-24, and the inhibitor-resistant TEM (IRT) TEM-44 and (ii) five novel enzymes, a penicillinase TEM-57 similar to TEM-1, an ESBL TEM-66 similar to TEM-3, and three IRTs, TEM-65, TEM-73, and TEM-74. The penicillinase TEM-57 and the ESBL TEM-66 differed from TEM-1 and TEM-3, respectively, by the amino acid substitution Gly-92-->Asp (nucleotide mutation G-477-->A). This substitution could have accounted for the decrease in pIs (5.2 for TEM-57 and 6.0 for TEM-66) but did not necessarily affect the intrinsic activities of these enzymes. The IRT TEM-65 was an IRT-1-like IRT (Cys-244) related to TEM-2 (Lys-39). The two other IRTs, TEM-73 and TEM-74, were related to IRT-1 (Cys-244) and IRT-2 (Ser-244), respectively, and harbored the amino acid substitutions Leu-21-->Phe and Thr-265-->Met. In this study, the ESBLs TEM-66, TEM-24, and TEM-3 were encoded by large (170- to 180-kb) conjugative plasmids that exhibited similar patterns after digestion and hybridization with the TEM and AAC(6')I probes. The three IRTs TEM-65, TEM-73, and TEM-74 were encoded by plasmids that ranged in size from 42 to 70 kb but for which no transfer was obtained. The characterization of five new plasmid-mediated TEM-type beta-lactamases and the first report of TEM-24 in P. mirabilis are evidence of the wide diversity of beta-lactamases produced in this species and of its possible role as a beta-lactamase-encoding plasmid reservoir.
Bonnet
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Laboratoire de Bactériologie, Faculté de Médecine, 63001 Clermont-Ferrand Cedex, France. Richard.Bonnet@u-clermont1.fr
De Champs
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Sirot
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Chanal
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Labia
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Sirot
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D000483:Alleles; D000658:Amoxicillin / Q000494:pharmacology; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D018014:Gene Transfer Techniques; D014644:Genetic Variation; D006824:Hybridization, Genetic; D007525:Isoelectric Focusing; D007700:Kinetics; D010403:Penicillin Resistance; D010641:Phenotype; D010957:Plasmids; D011513:Proteus mirabilis / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D017422:Sequence Analysis, DNA; D001618:beta-Lactamases / Q000737:chemistry / Q000235:genetics*
D004269:DNA, Bacterial; D000658:Amoxicillin; D001618:beta-Lactamases