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Journal of Bacteriology, February 2000, p. 1035-1045, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Insertional Inactivation of Genes Encoding
Components of the Sodium-Type Flagellar Motor and Switch of
Vibrio parahaemolyticus
Blaise R.
Boles and
Linda L.
McCarter*
Department of Microbiology, University of
Iowa, Iowa City, Iowa 52242
Received 18 August 1999/Accepted 24 November 1999
Vibrio parahaemolyticus possesses two types of
flagella, polar and lateral, powered by distinct energy sources, which
are derived from the sodium and proton motive forces, respectively. Although proton-powered flagella in Escherichia coli and
Salmonella enterica serovar Typhimurium have been
extensively studied, the mechanism of torque generation is still not
understood. Molecular knowledge of the structure of the sodium-driven
motor is only now being developed. In this work, we identify the switch
components, FliG, FliM, and FliN, of the sodium-type motor. This brings
the total number of genes identified as pertinent to polar motor
function to seven. Both FliM and FliN possess charged domains not found in proton-type homologs; however, they can interact with the
proton-type motor of E. coli to a limited extent. Residues
known to be critical for torque generation in the proton-type motor are
conserved in the sodium-type motor, suggesting a common mechanism for
energy transfer at the rotor-stator interface regardless of the driving force powering rotation. Mutants representing a complete panel of
insertionally inactivated switch and motor genes were constructed. All
of these mutants were defective in sodium-driven swimming motility.
Alkaline phosphatase could be fused to the C termini of MotB and MotY
without abolishing motility, whereas deletion of the unusual, highly
charged C-terminal domain of FliM disrupted motor function. All of the
mutants retained proton-driven, lateral motility over surfaces. Thus,
although central chemotaxis genes are shared by the polar and lateral
systems, genes encoding the switch components, as well as the motor
genes, are distinct for each motility system.
*
Corresponding author. Mailing address: Department of
Microbiology, University of Iowa, Iowa City, IA 52242. Phone: (319)
335-9721. Fax: (319) 335-7679. E-mail:
linda-mccarter{at}uiowa.edu.
Journal of Bacteriology, February 2000, p. 1035-1045, Vol. 182, No. 4
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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