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Journal of Bacteriology, June 2009, p. 3665-3676, Vol. 191, No. 11
0021-9193/09/$08.00+0     doi:10.1128/JB.01724-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Genetic Mapping of Secretion and Functional Determinants of the Vibrio cholerae TcpF Colonization Factor{triangledown}

Shelly J. Krebs, Thomas J. Kirn,{dagger} and Ronald K. Taylor*

Department of Microbiology and Immunology, Dartmouth Medical School, Hanover, New Hampshire 03755

Received 9 December 2008/ Accepted 14 March 2009

Colonization of the human small intestine by Vibrio cholerae requires the type IV toxin-coregulated pilus (TCP). TcpF, which is encoded within the tcp operon, is secreted from the bacterial cell by the TCP apparatus and is also essential for colonization. Bacteria lacking tcpF are deficient in colonization, and anti-TcpF antibodies are protective in the infant mouse cholera model. In order to elucidate the regions of the protein that are required for secretion through the TCP apparatus and for its function in colonization, random mutagenesis of tcpF was performed. Analysis of these mutants suggests that multiple regions throughout the protein influence extracellular secretion and that determinants near the C terminus are important for the function of TcpF in colonization. The TcpF proteins of certain environmental V. cholerae isolates with 31% to 66% identity to pathogenic V. cholerae TcpF showed higher similarity in regions identified as secretion determinants but diverged in regions found to be important for colonization. These environmental TcpF proteins are secreted from the pathogenic strain; however, they do not mediate colonization in the infant mouse model. Here we provide genetic evidence pointing toward regions of TcpF that influence secretion, as well as regions that play an important role in in vivo colonization.

* Corresponding author. Mailing address: Dartmouth Medical School, Department of Microbiology and Immunology, 7550 Vail Building, Hanover, NH 03755. Phone: (603) 650-1632. Fax: (603) 650-1318. E-mail: ronald.k.taylor{at}dartmouth.edu

{triangledown} Published ahead of print on 20 March 2009.

{dagger} Present address: Hospital of the University of Pennsylvania, Department of Pathology and Laboratory Medicine, 6 Founders, 3400 Spruce St., Philadelphia, PA 19104.

Journal of Bacteriology, June 2009, p. 3665-3676, Vol. 191, No. 11
0021-9193/09/$08.00+0     doi:10.1128/JB.01724-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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