Journal of Bacteriology, December 2003, p. 7068-7076, Vol. 185, No. 24
0021-9193/03/$08.00+0 DOI: 10.1128/JB.185.24.7068-7076.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
FimX, a Multidomain Protein Connecting Environmental Signals to Twitching Motility in Pseudomonas aeruginosa
Bixing Huang, Cynthia B. Whitchurch,
and John S. Mattick*
Institute
for Molecular Bioscience, University of Queensland, Brisbane, QLD 4072,
Australia
Received 18 June 2003/
Accepted 17 September 2003
Twitching
motility is a form of surface translocation mediated by the extension,
tethering, and retraction of type IV pili. Three independent
Tn5-B21 mutations of Pseudomonas aeruginosa with
reduced twitching motility were identified in a new locus which encodes
a predicted protein of unknown function annotated PA4959 in the P.
aeruginosa genome sequence. Complementation of these mutants with
the wild-type PA4959 gene, which we designated fimX, restored
normal twitching motility. fimX mutants were found to express
normal levels of pilin and remained sensitive to pilus-specific
bacteriophages, but they exhibited very low levels of surface pili,
suggesting that normal pilus function was impaired. The fimX
gene product has a molecular weight of 76,000 and contains four
predicted domains that are commonly found in signal transduction
proteins: a putative response regulator (CheY-like) domain, a PAS-PAC
domain (commonly involved in environmental sensing), and DUF1 (or
GGDEF) and DUF2 (or EAL) domains, which are thought to be involved in
cyclic di-GMP metabolism. Red fluorescent protein fusion experiments
showed that FimX is located at one pole of the cell via sequences
adjacent to its CheY-like domain. Twitching motility in fimX
mutants was found to respond relatively normally to a range of
environmental factors but could not be stimulated by tryptone and
mucin. These data suggest that fimX is involved in the
regulation of twitching motility in response to environmental
cues.
* Corresponding
author. Mailing address: Institute for Molecular Bioscience, University
of Queensland, Brisbane, QLD 4072, Australia. Phone: 61-7-3346-2110.
Fax: 61-7-3346-2111. E-mail:
j.mattick{at}imb.uq.edu.au.
Present
address: Department of Medicine, University of California, San
Francisco, San Francisco, CA 94143-0654.
Journal of Bacteriology, December 2003, p. 7068-7076, Vol. 185, No. 24
0021-9193/03/$08.00+0 DOI: 10.1128/JB.185.24.7068-7076.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Copyright © 2003 by the American Society for Microbiology. All rights reserved.