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Journal of Bacteriology, December 2002, p. 6566-6571, Vol. 184, No. 23
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.23.6566-6571.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Functional Analysis of the Heat Shock Regulator HrcA of Chlamydia trachomatis

Adam C. Wilson1 and Ming Tan1,2*

Department of Microbiology and Molecular Genetics,1 Department of Medicine, College of Medicine, University of California, Irvine, California 92697-40252

Received 18 June 2002/ Accepted 27 August 2002

HrcA is a regulator of bacterial heat shock gene expression that binds to a cis-acting DNA element called CIRCE. It has been proposed that HrcA and CIRCE function as a repressor-operator pair. We have purified recombinant HrcA from the pathogenic bacterium Chlamydia trachomatis and have shown that it is a DNA-binding protein that functions as a negative regulator of transcription. HrcA bound specifically to the CIRCE element in a concentration-dependent manner. HrcA repressed the in vitro transcription of a chlamydial heat shock promoter, and this repression was promoter specific. HrcA-mediated repression appears to be dependent on the topological state of the promoter, as repression on a supercoiled promoter template was greater than that on a linearized template. These results provide direct support for the role of HrcA as a transcriptional repressor in bacteria. This is the first report of the in vitro reconstitution of transcriptional regulation in Chlamydia.

* Corresponding author. Mailing address: B240 Med Sci I, Department of Microbiology and Molecular Genetics, University of California, Irvine, CA 92697-4025. Phone: (949) 824-3397. Fax: (949) 824-8598. E-mail: mingt{at}uci.edu.

Journal of Bacteriology, December 2002, p. 6566-6571, Vol. 184, No. 23
0021-9193/02/$04.00+0     DOI: 10.1128/JB.184.23.6566-6571.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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