This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Roberts, E. A.
Right arrow Articles by Friedman, R. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Roberts, E. A.
Right arrow Articles by Friedman, R. L.

 Previous Article  |  Next Article 

Journal of Bacteriology, August 2004, p. 5410-5417, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5410-5417.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of the eis Promoter of Mycobacterium tuberculosis

Esteban A. Roberts, Amanda Clark, Sarah McBeth, and Richard L. Friedman*

Department of Microbiology and Immunology, University of Arizona, Tucson, Arizona 85724

Received 19 April 2004/ Accepted 18 May 2004

To further understand Mycobacterium tuberculosis pathogenesis, the regulation of potential virulence genes needs to be investigated. The eis gene of M. tuberculosis H37Rv enhances the intracellular survival of Mycobacterium smegmatis, which does not contain eis, within macrophages (J. Wei, J. L. Dahl, J. W. Moulder, E. A. Roberts, P. O'Gaora, D. B. Young, and R. L. Friedman, J. Bacteriol. 182:377-384, 2000). Experiments were done to characterize the eis promoter in M. smegmatis and M. tuberculosis H37Ra. The putative –10 and –35 regions matched the Escherichia coli {sigma}70 consensus 67 and 83%, respectively, making it a group A/SigA-like mycobacterial promoter. Expression of site-directed variants of the core promoter region, determined by flow cytometry using gfp as a reporter, showed that the putative –10 region is essential for eis expression. In addition, site-directed alteration of the eis promoter to the consensus E. coli {sigma}70 promoter elements increased gfp transcription to levels similar to that driven by the heat shock promoter, phsp60, of Mycobacterium bovis BCG. Upstream promoter deletion analysis showed that a 200- and 412-bp region of the promoter was necessary for maximum expression of gfp in M. smegmatis and M. tuberculosis H37Ra, respectively. Random mutagenesis of the 412-bp eis promoter, using a catechol 2,3-dioxygenase screen and activity assay, defined nucleotides upstream of the core promoter region that are essential to eis expression in both M. smegmatis and M. tuberculosis H37Ra, including a region homologous to a DinR cis element.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, The University of Arizona College of Medicine, 1501 N. Campbell Ave., P.O. Box 245049, Tucson, AZ 85724. Phone: (520) 626-7807. Fax: (520) 626-2100. E-mail: rfriedma{at}email.arizona.edu.

Journal of Bacteriology, August 2004, p. 5410-5417, Vol. 186, No. 16
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.16.5410-5417.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wu, S., Barnes, P. F., Samten, B., Pang, X., Rodrigue, S., Ghanny, S., Soteropoulos, P., Gaudreau, L., Howard, S. T. (2009). Activation of the eis gene in a W-Beijing strain of Mycobacterium tuberculosis correlates with increased SigA levels and enhanced intracellular growth. Microbiology 155: 1272-1281 [Abstract] [Full Text]  
  • Vasudeva-Rao, H. M., McDonough, K. A. (2008). Expression of the Mycobacterium tuberculosis acr-Coregulated Genes from the DevR (DosR) Regulon Is Controlled by Multiple Levels of Regulation. Infect. Immun. 76: 2478-2489 [Abstract] [Full Text]  
  • Byrne, G. A., Russell, D. A., Chen, X., Meijer, W. G. (2007). Transcriptional Regulation of the virR Operon of the Intracellular Pathogen Rhodococcus equi. J. Bacteriol. 189: 5082-5089 [Abstract] [Full Text]  
  • Lella, R. K., Sharma, C. (2007). Eis (Enhanced Intracellular Survival) Protein of Mycobacterium tuberculosis Disturbs the Cross Regulation of T-cells. J. Biol. Chem. 282: 18671-18675 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»