Phosphorylation-Dependent Feedback Inhibition of RIG-I by DAPK1 Identified by Kinome-wide siRNA Screening

Mol Cell. 2017 Feb 2;65(3):403-415.e8. doi: 10.1016/j.molcel.2016.12.021. Epub 2017 Jan 26.


Cell-autonomous induction of type I interferon must be stringently regulated. Rapid induction is key to control virus infection, whereas proper limitation of signaling is essential to prevent immunopathology and autoimmune disease. Using unbiased kinome-wide RNAi screening followed by thorough validation, we identified 22 factors that regulate RIG-I/IRF3 signaling activity. We describe a negative-feedback mechanism targeting RIG-I activity, which is mediated by death associated protein kinase 1 (DAPK1). RIG-I signaling triggers DAPK1 kinase activation, and active DAPK1 potently inhibits RIG-I stimulated IRF3 activity and interferon-beta production. DAPK1 phosphorylates RIG-I in vitro at previously reported as well as other sites that limit 5'ppp-dsRNA sensing and virtually abrogate RIG-I activation.

Keywords: DAPK1; DDX58; RIG-I; antiviral response; cytokines; feedback regulation; innate immunity; interferon system; pattern recognition receptors; signal transduction.

MeSH terms

  • A549 Cells
  • Animals
  • Cells, Cultured
  • Death-Associated Protein Kinases / metabolism*
  • Feedback, Physiological
  • HEK293 Cells
  • Humans
  • Mice
  • Phosphorylation
  • Protein Kinases / metabolism
  • RNA, Small Interfering / genetics*
  • Receptors, Retinoic Acid / metabolism*
  • Signal Transduction


  • PLAAT4 protein, human
  • RNA, Small Interfering
  • Receptors, Retinoic Acid
  • Protein Kinases
  • DAPK1 protein, human
  • Death-Associated Protein Kinases