Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis.
Cyclic di-AMP is a recently discovered signaling molecule which regulates various aspects of bacterial physiology and virulence. Here we report the characterization of c-di-AMP synthesizing and hydrolyzing proteins from Mycobacterium tuberculosis. Recombinant Rv3586 (MtbDisA) can synthesize c-di-AMP...
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doaj-a50650623a474b2d82959b50928c94512020-11-25T02:33:35ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0191e8609610.1371/journal.pone.0086096Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis.Kasi ManikandanVaratharajan SabareeshNirpendra SinghKashyap SaigalUndine MecholdKrishna Murari SinhaCyclic di-AMP is a recently discovered signaling molecule which regulates various aspects of bacterial physiology and virulence. Here we report the characterization of c-di-AMP synthesizing and hydrolyzing proteins from Mycobacterium tuberculosis. Recombinant Rv3586 (MtbDisA) can synthesize c-di-AMP from ATP through the diadenylate cyclase activity. Detailed biochemical characterization of the protein revealed that the diadenylate cyclase (DAC) activity is allosterically regulated by ATP. We have identified the intermediates of the DAC reaction and propose a two-step synthesis of c-di-AMP from ATP/ADP. MtbDisA also possesses ATPase activity which is suppressed in the presence of the DAC 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. We have identified Rv2837c (MtbPDE) to have c-di-AMP specific phosphodiesterase activity. It hydrolyzes c-di-AMP to 5'-AMP in two steps. First, it linearizes c-di-AMP into pApA which is further hydrolyzed to 5'-AMP. 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. Our results suggest that the cellular concentration of c-di-AMP can be regulated by ATP concentration as well as the hydrolysis by MtbPDE.http://europepmc.org/articles/PMC3900455?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Kasi Manikandan Varatharajan Sabareesh Nirpendra Singh Kashyap Saigal Undine Mechold Krishna Murari Sinha |
spellingShingle |
Kasi Manikandan Varatharajan Sabareesh Nirpendra Singh Kashyap Saigal Undine Mechold Krishna Murari Sinha Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis. PLoS ONE |
author_facet |
Kasi Manikandan Varatharajan Sabareesh Nirpendra Singh Kashyap Saigal Undine Mechold Krishna Murari Sinha |
author_sort |
Kasi Manikandan |
title |
Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis. |
title_short |
Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis. |
title_full |
Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis. |
title_fullStr |
Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis. |
title_full_unstemmed |
Two-step synthesis and hydrolysis of cyclic di-AMP in Mycobacterium tuberculosis. |
title_sort |
two-step synthesis and hydrolysis of cyclic di-amp in mycobacterium tuberculosis. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2014-01-01 |
description |
Cyclic di-AMP is a recently discovered signaling molecule which regulates various aspects of bacterial physiology and virulence. Here we report the characterization of c-di-AMP synthesizing and hydrolyzing proteins from Mycobacterium tuberculosis. Recombinant Rv3586 (MtbDisA) can synthesize c-di-AMP from ATP through the diadenylate cyclase activity. Detailed biochemical characterization of the protein revealed that the diadenylate cyclase (DAC) activity is allosterically regulated by ATP. We have identified the intermediates of the DAC reaction and propose a two-step synthesis of c-di-AMP from ATP/ADP. MtbDisA also possesses ATPase activity which is suppressed in the presence of the DAC 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. We have identified Rv2837c (MtbPDE) to have c-di-AMP specific phosphodiesterase activity. It hydrolyzes c-di-AMP to 5'-AMP in two steps. First, it linearizes c-di-AMP into pApA which is further hydrolyzed to 5'-AMP. 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. Our results suggest that the cellular concentration of c-di-AMP can be regulated by ATP concentration as well as the hydrolysis by MtbPDE. |
url |
http://europepmc.org/articles/PMC3900455?pdf=render |
work_keys_str_mv |
AT kasimanikandan twostepsynthesisandhydrolysisofcyclicdiampinmycobacteriumtuberculosis AT varatharajansabareesh twostepsynthesisandhydrolysisofcyclicdiampinmycobacteriumtuberculosis AT nirpendrasingh twostepsynthesisandhydrolysisofcyclicdiampinmycobacteriumtuberculosis AT kashyapsaigal twostepsynthesisandhydrolysisofcyclicdiampinmycobacteriumtuberculosis AT undinemechold twostepsynthesisandhydrolysisofcyclicdiampinmycobacteriumtuberculosis AT krishnamurarisinha twostepsynthesisandhydrolysisofcyclicdiampinmycobacteriumtuberculosis |
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