1887

Abstract

MexXY, a drug efflux pump in , confers resistance to aminoglycoside antibiotics. We recently reported that MexZ binds to the promoter region of the operon. Electrophoretic mobility shift assay (EMSA) using recombinant MexZ and oligonucleotide probes prepared from the intergenic region between and revealed that MexZ binds to a 20 bp palindromic sequence. Culture of in the presence of tetracycline induced higher levels of MexX and MexZ, as measured by immunoblotting and EMSA, than in the absence of antibiotics. When MexZ was expressed by a expression plasmid, the plasmid-borne MexZ repressed drug-induced MexX production, further confirming that MexZ acts as a repressor of the operon. PA5471 protein has been reported to be essential for drug-induced MexXY production. Similarly to that report, we observed that plasmid-borne PA5471 induced both MexX and MexZ production in PAO1 cells. Interestingly, interaction between MexZ and PA5471 was observed in a yeast two-hybrid assay. Furthermore, EMSA and transcription assays revealed that interaction between PA5471 and MexZ reduced MexZ DNA-binding ability, leading to transcription. These findings contribute to the understanding of the molecular mechanisms underlying the transcriptional regulation of and by drug-induced PA5471 expression.

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2009-10-01
2024-12-07
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