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JAC Advance Access published online on September 14, 2005
Journal of Antimicrobial Chemotherapy, doi:10.1093/jac/dki333
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Published by Oxford University Press 2005
Received May 4, 2005
Revised July 27, 2005
Accepted August 17, 2005

Original article

Characterization of the MacA-MacB efflux system in Neisseria gonorrhoeae

Corinne E. Rouquette-Loughlin 1, Jacqueline T. Balthazar 1, and William M. Shafer 1*

1 Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, GA 30322, USA; Laboratories of Microbial Pathogenesis, VA Medical Center, Decatur, GA 30033, USA

* To whom correspondence should be addressed.
William M. Shafer, E-mail: wshafer{at}emory.edu


  Abstract

Objectives: A homologue of the MacA-MacB ABC transporter of Escherichia coli, which recognizes and exports macrolides, was identified in Neisseria gonorrhoeae. This study was undertaken to determine whether gonococci could use the MacA-MacB homologue to express decreased susceptibility to macrolides.

Methods: Techniques of DNA sequencing, gene cloning and expression of recombinant proteins in E. coli, gene mutation construction, transcriptional analysis and antimicrobial susceptibility testing were used in the study.

Results: Although the gonococcal MacA-MacB efflux pump enhanced bacterial resistance to macrolides when overexpressed in an E. coli background, its loss in a gonococcal clinical isolate only slightly decreased bacterial resistance to azithromycin and erythromycin. However, a mutation in the -10 sequence of the promoter used in macAB expression enhanced the macrolide resistance of gonococci that produced a defective MtrC-MtrD-MtrE pump, which also recognizes macrolides.

Conclusions: The results from this study indicate that gonococci can employ both the MacA-MacB and MtrC-MtrD-MtrE efflux pumps to develop resistance to macrolides, particularly if mutations develop in the promoter that drives transcription of macAB.

Keywords: gonococci; macrolides; resistance; efflux; regulation.
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