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Journal of Bacteriology, April 2005, p. 2681-2692, Vol. 187, No. 8
0021-9193/05/$08.00+0     doi:10.1128/JB.187.8.2681-2692.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Identification of Cyclic AMP-Regulated Genes in Mycobacterium tuberculosis Complex Bacteria under Low-Oxygen Conditions

Michaela A. Gazdik1 and Kathleen A. McDonough1,2*

Wadsworth Center,2 Department of Biomedical Sciences, University at Albany, Albany, New York1

Received 26 October 2004/ Accepted 10 January 2005

Mycobacterium tuberculosis is the etiological agent of tuberculosis (TB), which kills approximately 2 million people a year despite current treatment options. A greater understanding of the biology of this bacterium is needed to better combat TB disease. The M. tuberculosis genome encodes as many as 15 adenylate cyclases, suggesting that cyclic AMP (cAMP) has an important, yet overlooked, role in mycobacteria. This study examined the effect of exogenous cAMP on protein expression in Mycobacterium bovis BCG grown under hypoxic versus ambient conditions. Both shaking and shallow standing cultures were examined for each atmospheric condition. Different cAMP-dependent changes in protein expression were observed in each condition by two-dimensional gel electrophoresis. Shaking low-oxygen cultures produced the most changes (12), while standing ambient conditions showed the fewest (2). Five upregulated proteins, Rv1265, Rv2971, GroEL2, PE_PGRS6a, and malate dehydrogenase, were identified from BCG by mass spectrometry and were shown to also be regulated by cAMP at the mRNA level in both M. tuberculosis H37Rv and BCG. To our knowledge, these data provide the first direct evidence for cAMP-mediated gene regulation in TB complex mycobacteria.

* 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) 402-4773. E-mail: Kathleen.McDonough{at}wadsworth.org.

Journal of Bacteriology, April 2005, p. 2681-2692, Vol. 187, No. 8
0021-9193/05/$08.00+0     doi:10.1128/JB.187.8.2681-2692.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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