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Infection and Immunity, September 2003, p. 5332-5343, Vol. 71, No. 9
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.9.5332-5343.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

A Family of acr-Coregulated Mycobacterium tuberculosis Genes Shares a Common DNA Motif and Requires Rv3133c (dosR or devR) for Expression

Matthew A. Florczyk,1 Lee Ann McCue,1 Anjan Purkayastha,1,2 Egidio Currenti,1 Meyer J. Wolin,1 and Kathleen A. McDonough1,2*

Wadsworth Center, New York State Department of Health,1 Department of Biomedical Sciences, University at Albany, Albany, New York 122082

Received 19 December 2002/ Returned for modification 18 March 2003/ Accepted 24 June 2003

Previous work has shown that the divergently transcribed Mycobacterium tuberculosis genes acr (hspX, Rv2031c) and acg (Rv2032) are induced under conditions of shallow standing culture and low oxygen and intracellularly within macrophages. We used a combination of computational and experimental methods to identify promoters for eight additional genes that are regulated in a similar manner and that comprise an acr-coregulated promoter (ACP) family. Transcriptional regulation of these ACP family members was evaluated by using a plasmid-based promoter-green fluorescent protein fusion system and flow cytometry. All promoters showed increased expression in shallow standing versus shaking cultures, in low- versus high-oxygen conditions, and intracellularly within macrophages versus extracellularly in tissue culture medium. However, there were quantitative differences in expression among promoters and among conditions for each promoter. A conserved 18-bp palindromic sequence motif was identified in all ACPs by Gibbs sampling-based computational analyses. Two such motifs overlap regions in the acr and acg promoters that were previously shown to be required for their expression. In addition, we found that 5% carbon dioxide was required for growth of Mycobacterium bovis BCG under microaerophilic (1.3% O2) culture conditions and fully prevented the growth cessation typically associated with rapid removal of oxygen. These findings are likely to be relevant to the in vivo environment and will contribute to our understanding of the pathogenesis of tuberculosis infection.

* Corresponding author. Mailing address: Wadsworth Center, New York State Department of Health, P.O. Box 22002, 120 New Scotland Ave., Albany, NY 12208. Phone: (518) 486-4253. Fax: (518) 474-3181. E-mail: Kathleen.McDonough{at}wadsworth.org.

Editor: V. J. DiRita

Infection and Immunity, September 2003, p. 5332-5343, Vol. 71, No. 9
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.9.5332-5343.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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