The identification of the essential bacterial second messenger cyclic-di-AMP (c-di-AMP) synthesized by the DNA-integrity scanning protein A (DisA) has opened up a new and emerging field in bacterial signalling. To further analyse the diadenylate cyclase (DAC) reaction catalysed by the DAC domains of DisA, we crystallized Thermotoga maritima DisA in the presence of different ATP analogues and metal ions to identify the metal-binding site and trap the enzyme in pre- and post-reaction states. Through structural and biochemical assays we identified important residues essential for the reaction in the active site of the DAC domains. Our structures resolve the metal-binding site and thus explain the activation of ATP for the DAC reaction. Moreover, we were able to identify a potent inhibitor of the DAC domain. Based on the available structures and homology to annotated DAC domains we propose a common mechanism for c-di-AMP synthesis by DAC domains in c-di-AMP-producing species and a possible approach for its effective inhibition.
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Research Article|
July 23 2015
Structural analysis of the diadenylate cyclase reaction of DNA-integrity scanning protein A (DisA) and its inhibition by 3′-dATP
Martina Müller
;
Martina Müller
*Gene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, 81377 Munich, Germany
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Tobias Deimling
;
Tobias Deimling
*Gene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, 81377 Munich, Germany
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Karl-Peter Hopfner
;
Karl-Peter Hopfner
*Gene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, 81377 Munich, Germany
†Center for Integrated Protein Sciences, Ludwig-Maximilians-Universität, 81377 Munich, Germany
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Gregor Witte
Gregor Witte
1
*Gene Center and Department of Biochemistry, Ludwig-Maximilians-Universität, 81377 Munich, Germany
1To whom correspondence should be addressed (email witte@genzentrum.lmu.de).
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Biochem J (2015) 469 (3): 367–374.
Article history
Received:
March 25 2015
Revision Received:
May 20 2015
Accepted:
May 27 2015
Accepted Manuscript online:
May 27 2015
Citation
Martina Müller, Tobias Deimling, Karl-Peter Hopfner, Gregor Witte; Structural analysis of the diadenylate cyclase reaction of DNA-integrity scanning protein A (DisA) and its inhibition by 3′-dATP. Biochem J 1 August 2015; 469 (3): 367–374. doi: https://doi.org/10.1042/BJ20150373
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