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The helicase DDX41 recognizes the bacterial secondary messengers cyclic di-GMP and cyclic di-AMP to activate a type I interferon immune response

Abstract

The induction of type I interferons by the bacterial secondary messengers cyclic di-GMP (c-di-GMP) or cyclic di-AMP (c-di-AMP) is dependent on a signaling axis that involves the adaptor STING, the kinase TBK1 and the transcription factor IRF3. Here we identified the heliase DDX41 as a pattern-recognition receptor (PRR) that sensed both c-di-GMP and c-di-AMP. DDX41 specifically and directly interacted with c-di-GMP. Knockdown of DDX41 via short hairpin RNA in mouse or human cells inhibited the induction of genes encoding molecules involved in the innate immune response and resulted in defective activation of STING, TBK1 and IRF3 in response to c-di-GMP or c-di-AMP. Our results suggest a mechanism whereby c-di-GMP and c-di-AMP are detected by DDX41, which forms a complex with STING to signal to TBK1-IRF3 and activate the interferon response.

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Figure 1: The c-di-GMP- and c-di-AMP-mediated induction of the innate immune response in mouse DCs and human monocytes requires DDX41.
Figure 2: Cyclic dinucleotides activate interferon via DDX41 in primary cells.
Figure 3: DDX41 is a direct sensor of c-di-GMP.
Figure 4: The DDX41 DEAD-box domain is required for c-di-GMP- and c-di-AMP-mediated induction of IFN-β.
Figure 5: Both c-di-GMP and c-di-AMP require DDX41 for STING-dependent signaling.
Figure 6: The binding affinity of c-di-GMP for DDX41 is greater than that of c-di-GMP for STING.
Figure 7: DDX41 is required for the downstream association of c-di-GMP with STING.

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Acknowledgements

We thank K. Kelly-Scumpia for technical assistance; A. Dev for critical reading of the manuscript; and members of the Cheng and Modlin laboratories for discussion. Supported by the US National Institutes of Health (T32-AR058921 to K.P.; and P50 AR063020, R01 AI022553 and R01 AI073539, AI056154, AI067769, AI047868 and AR63020). Also supported in part by the Ministry of Science and Technology of China (2009DFB30310, 2013CB911103) and the National Natural Science Foundation of China (31070660, 31200559).

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Contributions

K.P., Z.Z., R.L.M., Y.L. and G.C. designed the research; K.P. and Z.Z. did and analyzed the biochemical experiments; R.M.T. did the confocal imaging and analysis; S.O., Y.J. and Z.-J.L. did the cloning, expression and purification of DDX41 and the STING carboxy-terminal domain; S.S.I. prepared peritoneal macrophages; M.S. prepared PBMCs; S.A.Z. did and analyzed experiments with primary cells; S.Z. and W.Z. did and analyzed the affinity capillary electrophoresis binding affinity experiments; and K.P. wrote the manuscript.

Corresponding authors

Correspondence to Yong-jun Liu or Genhong Cheng.

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The authors declare no competing financial interests.

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Parvatiyar, K., Zhang, Z., Teles, R. et al. The helicase DDX41 recognizes the bacterial secondary messengers cyclic di-GMP and cyclic di-AMP to activate a type I interferon immune response. Nat Immunol 13, 1155–1161 (2012). https://doi.org/10.1038/ni.2460

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