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Journal of Bacteriology, June 2004, p. 3296-3303, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3296-3303.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Secretion of Virulence Proteins from Campylobacter jejuni Is Dependent on a Functional Flagellar Export Apparatus

Michael E. Konkel,* John D. Klena, Vanessa Rivera-Amill, Marshall R. Monteville, Debabrata Biswas, Brian Raphael, and Joey Mickelson

School of Molecular Biosciences, Washington State University, Pullman, Washington 99164

Received 20 October 2003/ Accepted 19 February 2004

Campylobacter jejuni, a gram-negative motile bacterium, secretes a set of proteins termed the Campylobacter invasion antigens (Cia proteins). The purpose of this study was to determine whether the flagellar apparatus serves as the export apparatus for the Cia proteins. Mutations were generated in five genes encoding three structural components of the flagella, the flagellar basal body (flgB and flgC), hook (flgE2), and filament (flaA and flaB) genes, as well as in genes whose products are essential for flagellar protein export (flhB and fliI). While mutations that affected filament assembly were found to be nonmotile (Mot–) and did not secrete Cia proteins (S–), a flaA (flaB+) filament mutant was found to be nonmotile but Cia protein secretion competent (Mot–, S+). Complementation of a flaA flaB double mutant with a shuttle plasmid harboring either the flaA or flaB gene restored Cia protein secretion, suggesting that Cia export requires at least one of the two filament proteins. Infection of INT 407 human intestinal cells with the C. jejuni mutants revealed that maximal invasion of the epithelial cells required motile bacteria that are secretion competent. Collectively, these data suggest that the C. jejuni Cia proteins are secreted from the flagellar export apparatus.

* Corresponding author. Mailing address: School of Molecular Biosciences, Washington State University, Pullman, WA 99164. Phone: (509) 335-5039. Fax: (509) 335-1907. E-mail: konkel{at}mail.wsu.edu.

Journal of Bacteriology, June 2004, p. 3296-3303, Vol. 186, No. 11
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.11.3296-3303.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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