This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Tripathi, S. A.
Right arrow Articles by Taylor, R. K.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tripathi, S. A.
Right arrow Articles by Taylor, R. K.

 Previous Article  |  Next Article 

Journal of Bacteriology, June 2007, p. 4401-4409, Vol. 189, No. 12
0021-9193/07/$08.00+0     doi:10.1128/JB.00008-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Membrane Association and Multimerization of TcpT, the Cognate ATPase Ortholog of the Vibrio cholerae Toxin-Coregulated-Pilus Biogenesis Apparatus{triangledown}

Shital A. Tripathi and Ronald K. Taylor*

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

Received 2 January 2007/ Accepted 3 April 2007

The toxin-coregulated pilus (TCP) is one of the major virulence factors of Vibrio cholerae. Biogenesis of this type 4 pilus (Tfp) requires a number of structural components encoded by the tcp operon. TcpT, the cognate putative ATPase, is required for TCP biogenesis and all TCP-mediated functions. We studied the stability and localization of TcpT in cells containing in-frame deletions in each of the tcp genes. TcpT was detectable in each of the biogenesis mutants except the {Delta}tcpT strain. TcpT was localized to the inner membrane (IM) in a TcpR-dependent manner. TcpR is a predicted bitopic inner membrane protein of the TCP biogenesis apparatus. Using metal affinity pull-down experiments, we demonstrated interaction between TcpT and TcpR. Using Escherichia coli as a heterologous system, we investigated direct interaction between TcpR and TcpT. We report that TcpR is sufficient for TcpT IM localization per se; however, stable IM localization of TcpT requires an additional V. cholerae-specific factor(s). A LexA-based two-hybrid system was utilized to define interaction domains of the two proteins. We demonstrate a strong interaction between the cytoplasmic domain of TcpR and the N-terminal 100 amino acid residues of TcpT. We also demonstrated the ability of the C-terminal domain of TcpT to multimerize.

* Corresponding author. Mailing address: Dartmouth Medical School, Department of Microbiology and Immunology, HB7550, 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 13 April 2007.

Journal of Bacteriology, June 2007, p. 4401-4409, Vol. 189, No. 12
0021-9193/07/$08.00+0     doi:10.1128/JB.00008-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Martinez, R. M., Dharmasena, M. N., Kirn, T. J., Taylor, R. K. (2009). Characterization of Two Outer Membrane Proteins, FlgO and FlgP, That Influence Vibrio cholerae Motility. J. Bacteriol. 191: 5669-5679 [Abstract] [Full Text]  
  • Krebs, S. J., Kirn, T. J., Taylor, R. K. (2009). Genetic Mapping of Secretion and Functional Determinants of the Vibrio cholerae TcpF Colonization Factor. J. Bacteriol. 191: 3665-3676 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»