title: Mutagenesis mapping of the protein-protein interaction underlying FusB-type fusidic acid resistance.
abstract: FusB-type proteins represent the predominant mechanism of resistance to fusidic acid in staphylococci and act by binding to and modulating the function of the drug target (elongation factor G [EF-G]). To gain further insight into this antibiotic resistance mechanism, we sought to identify residues important for the interaction of FusB with EF-G and thereby delineate the binding interface within the FusB-EF-G complex. Replacement with alanine of any one of four conserved residues within the C-terminal domain of FusB (F156, K184, Y187, and F208) abrogated the ability of the protein to confer resistance to fusidic acid; the purified mutant proteins also lost the ability to bind S. aureus EF-G in vitro. E. coli EF-G, which is not ordinarily able to bind FusB-type proteins, was rendered competent for binding to FusB following deletion of a 3-residue tract (529SNP531) from domain IV of the protein. This study has identified key regions of both FusB and EF-G that are important for the interaction between the proteins, findings which corroborate our previous in silico prediction for the architecture of the complex formed between the resistance protein and the drug target (G. Cox, G. S. Thompson, H. T. Jenkins, F. Peske, A. Savelsbergh, M. V. Rodnina, W. Wintermeyer, S. W. Homans, T. A. Edwards, and A. J. O'Neill, Proc. Natl. Acad. Sci. U. S. A. 109:2102-2107, 2012).
authors.affiliation: Antimicrobial Research Centre and School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom.
authors.forename: Georgina
authors.initials: G
authors.lastname: Cox
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Edwards
authors.forename: Alex J
authors.initials: AJ
authors.lastname: O'Neill
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000187:drug effects; D005672:Fusidic Acid / Q000494:pharmacology*; D008826:Microbial Sensitivity Tests; D016296:Mutagenesis; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D013211:Staphylococcus aureus / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D005672:Fusidic Acid