The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses

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Abstract

Intracellular double-stranded RNA (dsRNA) is a chief sign of replication for many viruses. Host mechanisms detect the dsRNA and initiate antiviral responses. In this report, we identify retinoic acid inducible gene I (RIG-I), which encodes a DExD/H box RNA helicase that contains a caspase recruitment domain, as an essential regulator for dsRNA-induced signaling, as assessed by functional screening and assays. A helicase domain with intact ATPase activity was responsible for the dsRNA-mediated signaling. The caspase recruitment domain transmitted 'downstream' signals, resulting in the activation of transcription factors NF-κB and IRF-3. Subsequent gene activation by these factors induced antiviral functions, including type I interferon production. Thus, RIG-I is key in the detection and subsequent eradication of the replicating viral genomes.

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Figure 1: Identification of the CARD of RIG-I as a positive regulator for type I interferon.
Figure 2: The RIG-I CARD constitutively activates both IRF-3 and NF-κB.
Figure 3: Helicase activity is indispensable for signal transduction by RIG-I.
Figure 4: RIG-I is required for activation of IRF-3 and virus-induced gene expression.
Figure 5: Reduction in viral yield by RIG-I.
Figure 6: Dominant negative RIG-I inhibits NDV-induced but not TLR3-mediated activation of IRF-3.
Figure 7: Dominant negative RIG-I inhibits NDV-induced activation of IRF-3 in primary mouse fibroblasts.

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Acknowledgements

We thank E.L. Barsoumian for critical reading of the manuscript; S. Saito and M. Kohase for plaque assay protocol and suggestions; and M. Kohara for suggestions on real-time PCR. Supported by the Research for the Future Program; Japan Society for the Promotion of Science; Ministry of Education, Culture, Sports, Science and Technology of Japan; Nippon Boehringer Ingelheim; and Toray Industries.

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Correspondence to Takashi Fujita.

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Competing interests

The authors of this manuscript have filed a patent regarding RIG-I in collaboration with Toray Industries.

Supplementary information

Supplementary Fig. 1

IFN-treatment is prerequisite to virus-induced activation of IRF-3 in human K562 cells. (PDF 58 kb)

Supplementary Fig. 2

RIG-IC inhibits dsRNA-induced activation of reported gene expression. (PDF 28 kb)

Supplementary Fig. 3

Effect of anti-type I IFN antiserum on viral replication. (PDF 28 kb)

Supplementary Fig. 4

Expression of TBK1 in MEFs. (PDF 36 kb)

Supplementary Fig. 5

IRF-3 is not activated by polyU in mouse primary fibroblasts. (PDF 64 kb)

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Yoneyama, M., Kikuchi, M., Natsukawa, T. et al. The RNA helicase RIG-I has an essential function in double-stranded RNA-induced innate antiviral responses. Nat Immunol 5, 730–737 (2004) doi:10.1038/ni1087

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