Journal of Bacteriology, November 2001, p. 6636-6644, Vol. 183, No. 22
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.22.6636-6644.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Department of Medicine/Infectious Diseases, University of Florida, Gainesville, Florida 32610
Received 30 April 2001/Accepted 30 August 2001
Flagellar number in Pseudomonas aeruginosa is
controlled by FleN, a putative ATP/GTP binding protein. Disruption of
fleN results in multiflagellation of the otherwise
monoflagellate strains PAK and PAO1 and is associated with a
chemotactic defect. We propose that flagellar number is maintained by
the antiactivator FleN, which downregulates flagellar genes by binding
to their transcriptional activator, FleQ, an enhancer binding protein
belonging to the NifA subfamily. In this report we demonstrate direct
interaction of FleN and FleQ in the yeast two-hybrid system.
Mutagenesis of the putative ATP/GTP binding motif in
FleN24K
Q and truncation of FleN at either the N or C
terminus abrogates this interaction. FleN does not inhibit the DNA
binding ability of FleQ in vitro, thus indicating that it probably
utilizes another mechanism(s) to serve as a FleQ antiactivator.
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