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Journal of Bacteriology, August 2004, p. 5202-5209, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5202-5209.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

An Active Type IV Secretion System Encoded by the F Plasmid Sensitizes Escherichia coli to Bile Salts

Department of Biology, University of Central Oklahoma, Edmond, Oklahoma 73034,¹ Program in Molecular, Cell and Developmental Biology, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104²

Received 11 February 2004/ Accepted 5 May 2004

F⁺ strains of Escherichia coli infected with donor-specific bacteriophage such as M13 are sensitive to bile salts. We show here that this sensitivity has two components. The first derives from secretion of bacteriophage particles through the cell envelope, but the second can be attributed to expression of the F genes required for the formation of conjugative (F) pili. The latter component was manifested as reduced or no growth of an F⁺ strain in liquid medium containing bile salts at concentrations that had little or no effect on the isogenic F^– strain or as a reduced plating efficiency of the F⁺ strain on solid media; at 2% bile salts, plating efficiency was reduced 10⁴-fold. Strains with F or F-like R factors were consistently more sensitive to bile salts than isogenic, plasmid-free strains, but the quantitative effect of bile salts depended on both the plasmid and the strain. Sensitivity also depended on the bile salt, with conjugated bile salts (glycocholate and taurocholate) being less active than unconjugated bile salts (deoxycholate and cholate). F⁺ cells were also more sensitive to sodium dodecyl sulfate than otherwise isogenic F^– cells, suggesting a selectivity for amphipathic anions. A mutation in any but one F tra gene required for the assembly of F pili, including the traA gene encoding F pilin, substantially restored bile salt resistance, suggesting that bile salt sensitivity requires an active system for F pilin secretion. The exception was traW. A traW mutant was 100-fold more sensitive to cholate than the tra⁺ strain but only marginally more sensitive to taurocholate or glycocholate. Bile salt sensitivity could not be attributed to a generalized change in the surface permeability of F⁺ cells, as judged by the effects of hydrophilic and hydrophobic antibiotics and by leakage of periplasmic ß-lactamase into the medium.

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