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Journal of Bacteriology, March 2007, p. 1505-1513, Vol. 189, No. 5
0021-9193/07/$08.00+0     doi:10.1128/JB.01371-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Identification of Mycobacterial {sigma} Factor Binding Sites by Chromatin Immunoprecipitation Assays{triangledown}

Sébastien Rodrigue,1* Joëlle Brodeur,1 Pierre-Étienne Jacques,2 Alain L. Gervais,1 Ryszard Brzezinski,1 and Luc Gaudreau1*

Département de biologie, Université de Sherbrooke, 2500 boulevard de l'Université, Sherbrooke, Québec, Canada J1K 2R1,1 Institut de Recherches Cliniques de Montréal, 110 avenue des Pins Ouest, Montréal, Québec, Canada H2W 1R72

Received 29 August 2006/ Accepted 28 November 2006

Mycobacterium tuberculosis and Mycobacterium bovis are responsible for infections that cause a substantial amount of death, suffering, and loss around the world. Still, relatively little is known about the mechanisms of gene expression in these bacteria. Here, we used genome-wide location assays to identify direct target genes for mycobacterial {sigma} factors. Chromatin immunoprecipitation assays were performed with M. bovis BCG for Myc-tagged proteins expressed using an anhydrotetracycline-inducible promoter, and enriched DNA fragments were hybridized to a microarray representing intergenic regions from the M. tuberculosis H37Rv genome. Several putative target genes were validated by quantitative PCR. The corresponding transcriptional start sites were identified for {sigma}F, {sigma}C, and {sigma}K, and consensus promoter sequences are proposed. Our conclusions were supported by the results of in vitro transcription assays. We also examined the role of each holoenzyme in the expression of {sigma} factor genes. Our results revealed that many {sigma} factors are expressed from autoregulated promoters.

* Corresponding author. Mailing address: Département de biologie, Université de Sherbrooke, 2500 boulevard de l'Université, Sherbrooke, Québec, Canada J1K 2R1. Phone for Luc Gaudreau: (819) 821-8000, ext. 2082. Fax: (819) 821-8049. E-mail: Luc.Gaudreau{at}USherbrooke.ca. Phone for Sébastien Rodrigue: (819) 821-8000, ext. 2798. Fax: (819) 821-8049. E-mail: Sebastien.Rodrigue{at}USherbrooke.ca.

{triangledown} Published ahead of print on 8 December 2006.

Journal of Bacteriology, March 2007, p. 1505-1513, Vol. 189, No. 5
0021-9193/07/$08.00+0     doi:10.1128/JB.01371-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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