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Cyclic AMP intoxication of macrophages by a Mycobacterium tuberculosis adenylate cyclase

Abstract

With 8.9 million new cases and 1.7 million deaths per year, tuberculosis is a leading global killer that has not been effectively controlled1,2. The causative agent, Mycobacterium tuberculosis, proliferates within host macrophages where it modifies both its intracellular and local tissue environment, resulting in caseous granulomas with incomplete bacterial sterilization3,4. Although infection by various mycobacterial species produces a cyclic AMP burst within macrophages that influences cell signalling, the underlying mechanism for the cAMP burst remains unclear5,6,7. Here we show that among the 17 adenylate cyclase genes present in M. tuberculosis, at least one (Rv0386) is required for virulence. Furthermore, we demonstrate that the Rv0386 adenylate cyclase facilitates delivery of bacterial-derived cAMP into the macrophage cytoplasm. Loss of Rv0386 and the intramacrophage cAMP it delivers results in reductions in TNF-α production via the protein kinase A and cAMP response-element-binding protein pathway, decreased immunopathology in animal tissues, and diminished bacterial survival. Direct intoxication of host cells by bacterial-derived cAMP may enable M. tuberculosis to modify both its intracellular and tissue environments to facilitate its long-term survival.

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Figure 1: The M. tuberculosis -induced macrophage cAMP burst and CREB phosphorylation pathway require live bacilli but not host GPCR-AC.
Figure 2: Overexpression of phosphodiesterase Rv0805 in M. tuberculosis reduces intramacrophage cAMP levels, CREB phosphorylation, and TNF-α secretion in infected J774 cells.
Figure 3: The M. tuberculosisRv0386 mutant shows reduced mouse virulence, and lower levels of intramacrophage cAMP, CREB phosphorylation, and TNF-α secretion.
Figure 4: Bacterial-derived cAMP enters the macrophage cytosol after infection.

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Acknowledgements

The support of National Institutes of Health (NIH) awards AI30036, AI36973 and AI37856 is gratefully acknowledged.

Author Contributions N.A. and W.R.B. designed the research. N.A. performed the experiments. R.G., S.N. and G.L. designed and contributed to the mouse experiments. N.A. and W.R.B. analysed the data and wrote the paper.

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Correspondence to William R. Bishai.

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Agarwal, N., Lamichhane, G., Gupta, R. et al. Cyclic AMP intoxication of macrophages by a Mycobacterium tuberculosis adenylate cyclase. Nature 460, 98–102 (2009). https://doi.org/10.1038/nature08123

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