Identification of a Streptococcus pyogenes SF370 gene involved in production of c-di-AMP.

Taichi Kamegaya, Kenji Kuroda, Yoshihiro Hayakawa

Graduate School of Medicine, Nagoya University. kamegaya@med.nagoya-u.ac.jp

Nagoya journal of medical science 2011 Feb

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Here we show that bis(3'-5') cyclic diadenylic acid (c-di-AMP) and a diadenylate cyclase (DAC) domain protein involved in the biosynthesis of c-di-AMP were identified in Streptococcus pyogenes. The matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrum of the cell extract of S. pyogenes, which showed a fragment pattern very similar to that of the authentic sample of c-di-AMP, revealed that S. pyogenes produces c-di-AMP in the cell. Subsequently, we confirmed by an in vitro experiment that the production of c-di-AMP in the cell is due to the action of Spy1036 gene encoding a DAC domain protein named spyDAC, which is a new protein different from a well-known diadenylate cyclase. Moreover, the experiment gave a product with a molecular weight of 657.021, which is consistent with the molecular weight of c-di-AMP. Furthermore, the mass spectral fragment pattern of the product obtained by the in vitro biosynthesis is quite similar to that of the product produced by the above in vivo experiment. This in vitro production of c-di-AMP indicated that spyDAC in S. pyogenes actually catalyzes the in vivo biosynthesis of c-di-AMP from ATP.

Citation

Taichi Kamegaya, Kenji Kuroda, Yoshihiro Hayakawa. Identification of a Streptococcus pyogenes SF370 gene involved in production of c-di-AMP. Nagoya journal of medical science. 2011 Feb;73(1-2):49-57