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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.

Interaction of the Antiactivator FleN with the Transcriptional Activator FleQ Regulates Flagellar Number in Pseudomonas aeruginosa

Nandini Dasgupta and Reuben Ramphal*

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 FleN24Kright-arrow 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.

* Corresponding author. Mailing address: Department of Medicine/Infectious Diseases, P.O. Box 100277, JHMHC, University of Florida, Gainesville, FL 32610. Phone: (352) 392-2932. Fax: (352) 392-6481. E-mail: ramphr{at}medmac.ufl.edu.

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.


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