Dephosphorylation of the RNA sensors RIG-I and MDA5 by the phosphatase PP1 is essential for innate immune signaling

Immunity. 2013 Mar 21;38(3):437-49. doi: 10.1016/j.immuni.2012.11.018. Epub 2013 Mar 14.

Abstract

RIG-I and MDA5 have emerged as key cytosolic sensors for the detection of RNA viruses and lead to antiviral interferon (IFN) production. Recent studies have highlighted the importance of posttranslational modifications for controlling RIG-I antiviral activity. However, the regulation of MDA5 signal-transducing ability remains unclear. Here, we show that MDA5 signaling activity is regulated by a dynamic balance between phosphorylation and dephosphorylation of its caspase recruitment domains (CARDs). Employing a phosphatome RNAi screen, we identified PP1α and PP1γ as the primary phosphatases that are responsible for MDA5 and RIG-I dephosphorylation and that lead to their activation. Silencing of PP1α and PP1γ enhanced RIG-I and MDA5 CARD phosphorylation and reduced antiviral IFN-β production. PP1α- and PP1γ-depleted cells were impaired in their ability to induce IFN-stimulated gene expression, which resulted in enhanced RNA virus replication. This work identifies PP1α and PP1γ as regulators of antiviral innate immune responses to various RNA viruses, including influenza virus, paramyxovirus, dengue virus, and picornavirus.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Cells, Cultured
  • Chlorocebus aethiops
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases / genetics
  • DEAD-box RNA Helicases / immunology*
  • DEAD-box RNA Helicases / metabolism
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Immunity, Innate / genetics
  • Immunity, Innate / immunology*
  • Immunoblotting
  • Interferon-Induced Helicase, IFIH1
  • Interferon-beta / immunology
  • Interferon-beta / metabolism
  • Mice
  • Mice, Knockout
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Mutation
  • Phosphorylation
  • Protein Phosphatase 1 / genetics
  • Protein Phosphatase 1 / immunology*
  • Protein Phosphatase 1 / metabolism
  • RNA Interference
  • RNA, Viral / genetics
  • RNA, Viral / immunology*
  • RNA, Viral / metabolism
  • Signal Transduction / genetics
  • Signal Transduction / immunology
  • Vero Cells

Substances

  • RNA, Viral
  • Interferon-beta
  • Protein Phosphatase 1
  • DDX58 protein, human
  • IFIH1 protein, human
  • DEAD Box Protein 58
  • DEAD-box RNA Helicases
  • Interferon-Induced Helicase, IFIH1