Protein phosphatase PP1 negatively regulates the Toll-like receptor- and RIG-I-like receptor-triggered production of type I interferon by inhibiting IRF3 phosphorylation at serines 396 and 385 in macrophage

Cell Signal. 2014 Dec;26(12):2930-9. doi: 10.1016/j.cellsig.2014.09.007. Epub 2014 Sep 17.

Abstract

The production of type I interferon must be tightly regulated, and the aberrant production of this protein is harmful or even fatal to the host. The transcription factor IRF3 phosphorylation is a central regulator of type I interferon meditated antiviral response. Protein phosphatase-1 (PP1) has been reported to be important in many cell functions, including development, differentiation, and tumorigenesis. However, the roles of PP1 in Toll-like receptor (TLR)- or retinoic acid-inducible gene I like receptor (RLR)-triggered IRF-3 activation remain unclear. Here, we show that the activity of PP1 is downregulated in macrophages upon stimulation with TLR or RLR ligands, including lipopolysaccharide, and poly(I:C), or vesicular stomatitis virus (VSV), respectively. The overexpression of PP1 selectively inhibits TLR- and VSV-induced interferon regulatory factor 3 (IRF3) activation but has no substantial effect on TANK-binding kinase 1 (TBK1),ΚB kinase ε (IKKε) activation. Conversely, RNA interference of PP1 significantly promotes IRF3 activation. Consistently, The overexpression of PP1 inhibits TLR- and VSV-triggered IFN-β production while PP1 knockdown significantly increases the production of IFN-β in macrophages. We further demonstrate that PP1 directly interacts with IRF3 and dephosphorylates IRF3 at Ser385 and Ser396, resulting in the suppression of TLR- and RLR-triggered IFN-β production. Thus, PP1 functions as a negative feedback regulator of TLR- and RLR-triggered antiviral immune responses by acting as an IRF3 phosphatase.

Keywords: IFN-β; IRF3; PP1; RLR; TLR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Down-Regulation
  • Genes, Reporter
  • HEK293 Cells
  • Humans
  • Interferon Regulatory Factor-3 / metabolism*
  • Interferon-beta / metabolism*
  • Ligands
  • Macrophages / enzymology
  • Macrophages / metabolism*
  • Mice
  • Phosphorylation
  • Phosphoserine / metabolism*
  • Protein Binding
  • Protein Phosphatase 1 / metabolism*
  • Receptors, Cell Surface / metabolism*
  • Serine / metabolism
  • Toll-Like Receptors / metabolism*

Substances

  • Interferon Regulatory Factor-3
  • Irf3 protein, mouse
  • Ligands
  • Receptors, Cell Surface
  • Toll-Like Receptors
  • Phosphoserine
  • Serine
  • Interferon-beta
  • Protein Phosphatase 1