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Microbiology 152 (2006), 2405-2420; DOI  10.1099/mic.0.28860-0
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Microbiology 152 (2006), 2405-2420; DOI  10.1099/mic.0.28860-0
© 2006 Society for General Microbiology

Interaction and localization studies of enteropathogenic Escherichia coli type IV bundle-forming pilus outer membrane components

Anu Daniel{dagger}, Aparna Singh, Lynette J. Crowther, Paula J. Fernandes, Wiebke Schreiber and Michael S. Donnenberg

Division of Infectious Diseases, Department of Medicine, University of Maryland School of Medicine, 20 Penn Street, Baltimore, MD 21201, USA

Correspondence
Michael S. Donnenberg
mdonnenb{at}umaryland.edu

Typical enteropathogenic Escherichia coli strains express an established virulence factor belonging to the type IV pili family, called the bundle-forming pilus (BFP). BFP are present on the cell surface as bundled filamentous appendages, and are assembled and retracted by proteins encoded by the bfp operon. These proteins assemble to form a molecular machine. The BFP machine may be conceptually divided into three components: the cytoplasmic membrane (CM) subassembly, which is composed of CM proteins and cytoplasmic nucleotide-binding proteins; the outer membrane (OM) subassembly and the pilus itself. The authors have previously characterized the CM subassembly and the pilus. In this study, a more complete characterization of the OM subassembly was carried out using a combination of biochemical, biophysical and genetic approaches. It is reported that targeting of BfpG to the OM was influenced by the secretin BfpB. BfpG and BfpU interacted with the amino terminus of BfpB. BfpU had a complex cellular distribution pattern and, along with BfpB and BfpG, was part of the OM subassembly.

Abbreviations: AHT, anhydrotetracycline; AI, autoaggregation index; BFP, bundle-forming pilus; CM, cytoplasmic membrane; DSP, dithiobis[succinimidyl] propionate; EPEC, enteropathogenic Escherichia coli; {alpha}-Gal, {alpha}-galactosidase; His6, hexahistidine; ITC, isothermal titration calorimetry; MBP, maltose-binding protein; beta-ME, beta-mercaptoethanol; OM, outer membrane; Tfp, type IV pili

{dagger}Present address: Laboratory of Bacterial Pathogenesis and Immunology, The Rockefeller University, 1230 York Avenue, NY 10021, USA.




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