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Journal of Bacteriology, December 2006, p. 8479-8486, Vol. 188, No. 24
0021-9193/06/$08.00+0     doi:10.1128/JB.00157-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Effect of Site-Specific Mutations in Different Phosphotransfer Domains of the Chemosensory Protein ChpA on Pseudomonas aeruginosa Motility ^,

Andrew J. Leech and John S. Mattick^*

Institute for Molecular Bioscience, University of Queensland, Brisbane, Queensland 4072, Australia

Received 28 January 2006/ Accepted 19 September 2006

The virulence of Pseudomonas aeruginosa and other surface pathogens involves the coordinate expression of a wide range of virulence determinants, including type IV pili. These surface filaments are important for the colonization of host epithelial tissues and mediate bacterial attachment to, and translocation across, surfaces by a process known as twitching motility. This process is controlled in part by a complex signal transduction system whose central component, ChpA, possesses nine potential sites of phosphorylation, including six histidine-containing phosphotransfer (HPt) domains, one serine-containing phosphotransfer domain, one threonine-containing phosphotransfer domain, and one CheY-like receiver domain. Here, using site-directed mutagenesis, we show that normal twitching motility is entirely dependent on the CheY-like receiver domain and partially dependent on two of the HPt domains. Moreover, under different assay conditions, point mutations in several of the phosphotransfer domains of ChpA give rise to unusual "swarming" phenotypes, possibly reflecting more subtle perturbations in the control of P. aeruginosa motility that are not evident from the conventional twitching stab assay. Together, these results suggest that ChpA plays a central role in the complex regulation of type IV pilus-mediated motility in P. aeruginosa.

Published ahead of print on 29 September 2006.

Supplemental material for this article may be found at http://jb.asm.org/.

Present address: Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298-0678.

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