Duplication of the chromosomal blaSHV-11 gene in a clinical hypermutable strain of Klebsiella pneumoniae.
In a collection of 110 clinical isolates of Klebsiella pneumoniae, a single strain, Kp593, was found to exhibit a mutator phenotype with a rifampicin mutation frequency 100-fold higher than the modal value for this species. Complementation experiments with the wild-type MutL, one of the main components of the methyl-directed mismatch repair system, allowed the mutator phenotype to be reversed. Sequencing revealed substitution of the conserved residue Lys307 to Arg and site-directed mutagenesis followed by complementation experiments confirmed the critical role of this mutation. The patient infected with Kp593 relapsed a month later and the strain isolated then, Kp869, was identical to Kp593, as verified by PFGE analysis. Phenotypically, Kp869 colonies were more mucoid than those of Kp593, probably due to increased capsule synthesis as shown by electron microscopy. In addition, Kp869 exhibited a 16-fold higher amoxicillin resistance level related to a 36.4&emsp14;kb tandem duplication encompassing the chromosomal bla(SHV-11) gene, which was unstable in vitro. These data suggest that the mutator phenotype found in Kp593/Kp869 is associated with beneficial mutations conferring a selective advantage, such as increased virulence factor production and antibiotic resistance. The latter was due to resistance gene duplication, an event rarely described in natural isolates. This is the first description of the in vivo occurrence of gene duplication in a mutator background.
Duvernay
Chloé
C
CNRS UMR 5234, Laboratoire de Microbiologie Cellulaire et Moléculaire et Pathogénicité, Université Victor Segalen Bordeaux 2, Bordeaux, France.
Coulange
Laure
L
CNRS UMR 5234, Laboratoire de Microbiologie Cellulaire et Moléculaire et Pathogénicité, Université Victor Segalen Bordeaux 2, Bordeaux, France.
Dutilh
Brigitte
B
Laboratoire d'Analyses Médicales, Bordeaux, France.
Dubois
Véronique
V
CNRS UMR 5234, Laboratoire de Microbiologie Cellulaire et Moléculaire et Pathogénicité, Université Victor Segalen Bordeaux 2, Bordeaux, France.
Quentin
Claudine
C
CNRS UMR 5234, Laboratoire de Microbiologie Cellulaire et Moléculaire et Pathogénicité, Université Victor Segalen Bordeaux 2, Bordeaux, France.
Arpin
Corinne
C
CNRS UMR 5234, Laboratoire de Microbiologie Cellulaire et Moléculaire et Pathogénicité, Université Victor Segalen Bordeaux 2, Bordeaux, France.
D000369:Aged, 80 and over; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005260:Female; D020440:Gene Duplication; D005816:Genetic Complementation Test; D006801:Humans; D007710:Klebsiella Infections / Q000382:microbiology; D007711:Klebsiella pneumoniae / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D008826:Microbial Sensitivity Tests; D051719:MutS DNA Mismatch-Binding Protein / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010641:Phenotype; D017422:Sequence Analysis, DNA; D014774:Virulence; D001618:beta-Lactamases / Q000235:genetics*; D047090:beta-Lactams / Q000494:pharmacology
D004269:DNA, Bacterial; D047090:beta-Lactams; C107197:beta-lactamase SHV-11; D001618:beta-Lactamases; D051719:MutS DNA Mismatch-Binding Protein