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Journal of Bacteriology, July 2008, p. 4437-4446, Vol. 190, No. 13
0021-9193/08/$08.00+0     doi:10.1128/JB.00055-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

The Hybrid Sensor Kinase RscS Integrates Positive and Negative Signals To Modulate Biofilm Formation in Vibrio fischeri{triangledown}

Kati Geszvain{dagger} and Karen L. Visick*

Department of Microbiology and Immunology, Loyola University Chicago, Maywood, Illinois 60153

Received 11 January 2008/ Accepted 16 April 2008

Overexpression of the Vibrio fischeri sensor kinase RscS induces expression of the syp (symbiosis polysaccharide) gene cluster and promotes biofilm phenotypes such as wrinkled colony morphology, pellicle formation, and surface adherence. RscS is predicted to be a hybrid sensor kinase with a histidine kinase/ATPase (HATPase) domain, a receiver (Rec) domain, and a histidine phosphotransferase (Hpt) domain. Bioinformatic analysis also revealed the following three potential signal detection domains within RscS: two transmembrane helices forming a transmembrane region (TMR), a large periplasmic (PP) domain, and a cytoplasmic PAS domain. In this work, we genetically dissected the contributions of these domains to RscS function. Substitutions within the carboxy-terminal domain supported identification of RscS as a hybrid sensor kinase; disruption of both the HATPase and Rec domains eliminated induction of syp transcription, wrinkled colony morphology, pellicle formation, and surface adherence, while disruption of Hpt resulted in decreased activity. The PAS domain was also critical for RscS activity; substitutions in PAS resulted in a loss of activity. Generation of a cytoplasmic, N-terminal deletion derivative of RscS resulted in a partial loss of activity, suggesting a role for localization to the membrane and/or sequences within the TMR and PP domain. Finally, substitutions within the first transmembrane helix of the TMR and deletions within the PP domain both resulted in increased activity. Thus, RscS integrates both inhibitory and stimulatory signals from the environment to regulate biofilm formation by V. fischeri.


* Corresponding author. Mailing address: Department of Microbiology and Immunology, Loyola University Chicago, 2160 S. First Ave., Bldg. 105, Maywood, IL 60153. Phone: (708) 216-0869. Fax: (708) 216-9574. E-mail: kvisick{at}lumc.edu

{triangledown} Published ahead of print on 25 April 2008.

{dagger} Present address: Department of Environmental and Biomolecular Systems, Oregon Health and Sciences University, 20000 NW Walker Rd., Beaverton, OR 97006-8921.


Journal of Bacteriology, July 2008, p. 4437-4446, Vol. 190, No. 13
0021-9193/08/$08.00+0     doi:10.1128/JB.00055-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.




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