The projects MLP6201, FAC002, NWP036 and NWP004 of Council of Scientific and Industrial Research are gratefully acknowledged for generous funding to IGIB. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.įunding: The work was supported by a grant from Department of Biotechnology, Govt. Received: AugAccepted: DecemPublished: January 23, 2014Ĭopyright: © 2014 Manikandan et al. PLoS ONE 9(1):Įditor: Narayanaswamy Srinivasan, Indian Institute of Science, India Our results suggest that the cellular concentration of c-di-AMP can be regulated by ATP concentration as well as the hydrolysis by MtbPDE.Ĭitation: Manikandan K, Sabareesh V, Singh N, Saigal K, Mechold U, Sinha KM (2014) Two-Step Synthesis and Hydrolysis of Cyclic di-AMP in Mycobacterium tuberculosis. MtbPDE is novel compared to c-di-AMP specific phosphodiesterase, YybT (or GdpP) in being a soluble protein and hydrolyzing c-di-AMP to 5′-AMP. First, it linearizes c-di-AMP into pApA which is further hydrolyzed to 5′-AMP. It hydrolyzes c-di-AMP to 5′-AMP in two steps. We have identified Rv2837c (MtbPDE) to have c-di-AMP specific phosphodiesterase activity. Investigations by liquid chromatography -electrospray ionization mass spectrometry have detected multimeric forms of c-di-AMP which have implications for the regulation of c-di-AMP cellular concentration and various pathways regulated by the dinucleotide. MtbDisA also possesses ATPase activity which is suppressed in the presence of the DAC activity. We have identified the intermediates of the DAC reaction and propose a two-step synthesis of c-di-AMP from ATP/ADP. Detailed biochemical characterization of the protein revealed that the diadenylate cyclase (DAC) activity is allosterically regulated by ATP. Recombinant Rv3586 (MtbDisA) can synthesize c-di-AMP from ATP through the diadenylate cyclase activity. Here we report the characterization of c-di-AMP synthesizing and hydrolyzing proteins from Mycobacterium tuberculosis. Cyclic di-AMP is a recently discovered signaling molecule which regulates various aspects of bacterial physiology and virulence.
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