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J Biol Chem, Vol. 273, Issue 29, 18665-18673, July 17, 1998
From the School of Biological Sciences, University of Sydney,
Sydney, New South Wales 2006, Australia
The Staphylococcus aureus QacA
protein is a multidrug transporter that confers resistance to a broad
range of antimicrobial agents via proton motive
force-dependent efflux of the compounds. Primer extension
analysis was performed to map the transcription start points of the
qacA and divergently transcribed qacR
mRNAs. Each gene utilized a single promoter element, the locations
of which were confirmed by site-directed mutagenesis. Fusions of the
qacA and qacR promoters to a chloramphenicol
acetyl transferase reporter gene were used to demonstrate that QacR is
a trans-acting repressor of qacA transcription
that does not autoregulate its own expression. An inverted repeat
overlapping the qacA transcription start site was shown to
be the operator sequence for control of qacA gene
expression. Removal of one half of the operator prevented QacR-mediated
repression of the qacA promoter. Purified QacR protein bound specifically to this operator sequence in DNase I-footprinting experiments. Importantly, addition of diverse QacA substrates was shown
to induce qacA expression in vivo, as well as
inhibit binding of QacR to operator DNA in vitro, by using
gel-mobility shift assays. QacR therefore appears to interact directly
with structurally dissimilar inducing compounds that are substrates of
the QacA multidrug efflux pump.
QacR Is a Repressor Protein That Regulates Expression of the
Staphylococcus aureus Multidrug Efflux Pump QacA
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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