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Journal of Bacteriology, August 2009, p. 5304-5311, Vol. 191, No. 16
0021-9193/09/$08.00+0     doi:10.1128/JB.00551-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Site-Directed Mutagenesis Identifies a Molecular Switch Involved in Copper Sensing by the Histidine Kinase CinS in Pseudomonas putida KT2440{triangledown} ,§

Davide Quaranta,1 Megan M. McEvoy,2 and Christopher Rensing1*

Department of Soil, Water, and Environmental Science,1 Department of Chemistry and Biochemistry, University of Arizona, Tucson, Arizona 857212

Received 24 April 2009/ Accepted 5 June 2009

In the presence of copper, Pseudomonas putida activates transcription of cinAQ via the two-component system CinS-CinR. The CinS-CinR TCS was responsive to 0.5 µM copper and was specifically activated only by copper and silver. Modeling studies of CinS identified a potential copper binding site containing H37 and H147. CinS mutants with H37R and H147R mutations had an almost 10-fold reduced copper-dependent induction of cinAQ compared to the wild type.

* Corresponding author. Mailing address: Department of Soil, Water, and Environmental Science, University of Arizona, Shantz Bld #38 Rm 429, Tucson, AZ 85721. Phone: (520) 626-8482. Fax: (520) 621-1647. E-mail: rensingc{at}ag.arizona.edu

{triangledown} Published ahead of print on 19 June 2009.

§ Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, August 2009, p. 5304-5311, Vol. 191, No. 16
0021-9193/09/$08.00+0     doi:10.1128/JB.00551-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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