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A phosphomimetic-based mechanism of dengue virus to antagonize innate immunity

Nature Immunology volume 17, pages 523530 (2016) | Download Citation

Abstract

14-3-3 proteins regulate biological processes by binding to phosphorylated serine or phosphorylated threonine motifs of cellular proteins. Among the 14-3-3 proteins, 14-3-3ɛ serves a crucial function in antiviral immunity by mediating the cytosol–to–mitochondrial membrane translocation of the pathogen sensor RIG-I. Here we found that the NS3 protein of dengue virus (DV) bound to 14-3-3ɛ and prevented translocation of RIG-I to the adaptor MAVS and thereby blocked antiviral signaling. Intriguingly, a highly conserved phosphomimetic RxEP motif in NS3 was essential for the binding of 14-3-3ɛ. A recombinant mutant DV deficient in binding to 14-3-3ɛ showed impairment in antagonism of RIG-I and elicited a markedly augmented innate immune response and enhanced T cell activation. Our work reveals a novel phosphomimetic-based mechanism for viral antagonism of 14-3-3-mediated immunity, which might guide the rational design of therapeutics.

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Acknowledgements

We thank C. Huang (Center for Disease Control and Prevention) for the DV2 16681 infectious clone pD2/IC-30P; E. Harris (University of California, Berkeley) for antibody E1D8 to NS3; M. Diamond (Washington University in St. Louis) for cDNA encoding NS3 of WNV strain NY99 or Kunjin; Z. Chen (University of Texas Southwestern Medical Center) for cDNA encoding NS3 of HCV strain Con1 and plasmids encoding the HCV NS3-NS4A protease complex and its S139A catalytically-inactive mutant; R. Kuhn (Purdue University) for cDNA encoding NS3 of YFV strain 17D; J. Jung (University of Southern California) for Huh7.5 cells; M. Farzan (Scripps Institute Florida) for Huh7, BHK-21, C6/36 and K562 cells; N. Hacohen (Massachusetts General Hospital) for HEK293T ISRE-luciferase cells, L. Gehrke (Harvard University) for DV4 strain 814699; D. Knipe (Harvard University) for herpes simplex virus type 1; S. Whelan (Harvard University) for VSV-GFP; and A. García-Sastre (Icahn School of Medicine at Mount Sinai) for NDV-GFP. Supported by the US National Institutes of Health (R01 AI087846), the Giovanni Armenise-Harvard Foundation, the William F. Milton Fund, and a John and Virginia Kaneb Fellowship (M.U.G.).

Author information

Affiliations

  1. Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.

    • Ying Kai Chan
    •  & Michaela U Gack
  2. Department of Microbiology, The University of Chicago, Chicago, Illinois, USA.

    • Michaela U Gack

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Contributions

Y.K.C. performed all aspects of this study; and Y.K.C. and M.U.G. designed the study and wrote the manuscript.

Competing interests

The authors are co-inventors on a patent application for use of the RxEP motif and DV2(KIKP) mutant virus.

Corresponding author

Correspondence to Michaela U Gack.

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DOI

https://doi.org/10.1038/ni.3393

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