title: Interactions of the QacR multidrug-binding protein with structurally diverse ligands: implications for the evolution of the binding pocket.
abstract: The QacR multidrug-binding repressor protein regulates the expression of the Staphylococcus aureus qacA gene, a multidrug resistance (MDR) locus that is prevalent in clinical isolates of this important human pathogen. In this paper we demonstrate that the range of structurally diverse compounds capable of inducing qacA transcription is significantly more varied than previously appreciated, particularly in relation to bivalent cations. For all of the newly identified inducing compounds, induction of qacAexpression was correlated with a matching ability to dissociate QacR from operator DNA. Development of a ligand-binding assay based on intrinsic tryptophan fluorescence permitted dissociation constants to be determined for the majority of known QacR ligands, with values ranging from 0.2 to 82 microM. High-affinity binding of a compound to QacR in vitro was not found to correlate very strongly with either its in vivo inducing abilities or its structure. The latter observation indicated that the QacR ligand-binding pocket appears to have evolved to accommodate a wide range of toxic hydrophobic cations, rather than a specific class of compound. Importantly, the antimicrobial ligands of QacR included several plant alkaloids that share structural similarities with synthetic MDR substrates. This is consistent with the suggestion that the qacA-qacR MDR locus was recently derived from genes that protect against natural antimicrobial compounds.
authors.affiliation: School of Biological Sciences, University of Sydney, Sydney, New South Wales 2006, Australia.
authors.forename: Steve
authors.initials: S
authors.lastname: Grkovic
authors.forename: Kate M
authors.initials: KM
authors.lastname: Hardie
authors.forename: Melissa H
authors.initials: MH
authors.lastname: Brown
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Skurray
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D024901:Drug Resistance, Multiple, Bacterial / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism; D019143:Evolution, Molecular; D015964:Gene Expression Regulation, Bacterial; D008024:Ligands; D026901:Membrane Transport Proteins / Q000096:biosynthesis / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D011815:R Factors / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013211:Staphylococcus aureus / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D014364:Tryptophan / Q000737:chemistry
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D008024:Ligands; D026901:Membrane Transport Proteins; C442128:QacR protein, Staphylococcus aureus; D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; C068498:qacA protein, Staphylococcus aureus; D014364:Tryptophan