LmrR-mediated gene regulation of multidrug resistance in Lactococcus lactis

Abstract

Multidrug resistance (MDR) in Lactococcus lactis is due to the expression of the membrane ATP-binding cassette (ABC) transporter LmrCD. In the absence of drugs, the transcriptional regulator LmrR prevents expression of the lmrCD operon by binding to its operator site. Through an autoregulatory mechanism LmrR also suppresses its own expression. Although the lmrR and lmrCD genes have their own promoters, primer extension analysis showed the presence of a long transcript spanning the entire lmrR–lmrCD cluster, in addition to various shorter transcripts harbouring the lmrCD genes only. ‘In-gel’ Cu-phenanthroline footprinting analysis indicated an extensive interaction between LmrR and the lmrR promoter/operator region. Atomic force microscopy imaging of the binding of LmrR to the control region of lmrR DNA showed severe deformations indicative of DNA wrapping and looping, while LmrR binding to a fragment containing the lmrCD control region induced DNA bending. The results further suggest a drug-dependent regulation mechanism in which the lmrCD genes are co-transcribed with lmrR as a polycistronic messenger. This leads to an LmrR-mediated regulation of lmrCD expression that is exerted from two different locations and by distinct regulatory mechanisms.