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Journal of Bacteriology, November 2006, p. 7364-7377, Vol. 188, No. 21
0021-9193/06/$08.00+0     doi:10.1128/JB.01014-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Selective Promoter Recognition by Chlamydial {sigma}28 Holoenzyme{triangledown} ,{dagger}

Li Shen,1* Xiaogeng Feng,1 Yuan Yuan,2 Xudong Luo,1 Thomas P. Hatch,3 Kelly T. Hughes,4 Jun S. Liu,2 and You-xun Zhang1

Department of Medicine, Boston Medical Center, Boston University School of Medicine, Boston, Massachusetts 02118,1 Department of Statistics, Harvard University, Cambridge, Massachusetts 02138,2 Department of Molecular Sciences, University of Tennessee, Memphis, Tennessee 38163,3 Department of Biology, University of Utah, Salt Lake City, Utah 841124

Received 11 July 2006/ Accepted 15 August 2006

The {sigma} transcription factor confers the promoter recognition specificity of RNA polymerase (RNAP) in eubacteria. Chlamydia trachomatis has three known sigma factors, {sigma}66, {sigma}54, and {sigma}28. We developed two methods to facilitate the characterization of promoter sequences recognized by C. trachomatis {sigma}28 ({sigma}28Ct). One involved the arabinose-induced expression of plasmid-encoded {sigma}28Ct in a strain of Escherichia coli defective in the {sigma}28 structural gene, fliA. The second was an analysis of transcription in vitro with a hybrid holoenzyme reconstituted with E. coli RNAP core and recombinant {sigma}28Ct. These approaches were used to investigate the interactions of {sigma}28Ct with the {sigma}28Ct-dependent hctB promoter and selected E. coli {sigma}28 ({sigma}28Ec)-dependent promoters, in parallel, compared with the promoter recognition properties of {sigma}28EC. Our results indicate that RNAP containing {sigma}28Ct has at least three characteristics: (i) it is capable of recognizing some but not all {sigma}28EC-dependent promoters; (ii) it can distinguish different promoter structures, preferentially activating promoters with upstream AT-rich sequences; and (iii) it possesses a greater flexibility than {sigma}28EC in recognizing variants with different spacing lengths separating the –35 and –10 elements of the core promoter.

* Corresponding author. Mailing address: Department of Medicine, Boston Medical Center, Boston University School of Medicine, Boston, MA 02118. Phone: (617) 414-5278. Fax: (617) 414-5280. E-mail: lshen{at}bu.edu.

{triangledown} Published ahead of print on 25 August 2006.

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

Journal of Bacteriology, November 2006, p. 7364-7377, Vol. 188, No. 21
0021-9193/06/$08.00+0     doi:10.1128/JB.01014-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

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