A mechanism of carbapenem resistance due to a new insertion element (ISPa133) in Pseudomonas aeruginosa.
This study explored the evolutionary mechanism by which the clinical isolate PA110514 yields the imipenemresistant derivative PA116136. Both isolates were examined by PFGE and SDS-PAGE, which led to the identification of a new insertion sequence, ISPa133. This element was shown to have distinct chromosomal locations in each of the original isolates that appeared to explain the differences in imipenem susceptibilty. In strain PA110514, ISPa133 is located 56 nucleotides upstream of the translational start codon, which has no effect on expression of the porin OprD. However, in strain PA116136 ISPa133 it is located in front of nucleotide 696 and, by interrupting the coding region, causes a loss of OprD expression, thus conferring imipenem resistance. In vitro experiments mimicking the natural conditions of selective pressure yielded imipenem-resistant strains in which ISPa133 similarly interrupted oprD. A mechanism is proposed whereby ISPa133 acts as a mobile switch, with its position in oprD depending on the degree of selective pressure exerted by imipenem.
Molecular Microbiology and Antimicrobials Laboratory, Department of Pathology and Experimental Therapeutics, Medical School, IDIBELL, University of Barcelona, L'Hospitalet, Barcelona, Spain.
Lidia
L
Ruiz-Martínez
Lidia
L
López-Jiménez
Valeria
V
d'Ostuni
Ester
E
Fusté
Teresa
T
Vinuesa
Miguel
M
Viñas
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015780:Carbapenems / Q000494:pharmacology*; D018387:Codon, Initiator; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D020543:Proteome / Q000032:analysis; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA; D018440:beta-Lactam Resistance
D000900:Anti-Bacterial Agents; D015780:Carbapenems; D018387:Codon, Initiator; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D020543:Proteome