Lck/Hck/Fgr-Mediated Tyrosine Phosphorylation Negatively Regulates TBK1 to Restrain Innate Antiviral Responses

Cell Host Microbe. 2017 Jun 14;21(6):754-768.e5. doi: 10.1016/j.chom.2017.05.010.

Abstract

Cytosolic nucleic acid sensing elicits interferon production for primary antiviral defense through cascades controlled by protein ubiquitination and Ser/Thr phosphorylation. Here we show that TBK1, a core kinase of antiviral pathways, is inhibited by tyrosine phosphorylation. The Src family kinases (SFKs) Lck, Hck, and Fgr directly phosphorylate TBK1 at Tyr354/394, to prevent TBK1 dimerization and activation. Accordingly, antiviral sensing and resistance were substantially enhanced in Lck/Hck/Fgr triple knockout cells and ectopic expression of Lck/Hck/Fgr dampened the antiviral defense in cells and zebrafish. Small-molecule inhibitors of SFKs, which are conventional anti-tumor therapeutics, enhanced antiviral responses and protected zebrafish and mice from viral attack. Viral infection induced the expression of Lck/Hck/Fgr through TBK1-mediated mobilization of IRF3, thus constituting a negative feedback loop. These findings unveil the negative regulation of TBK1 via tyrosine phosphorylation and the functional integration of SFKs into innate antiviral immunity.

Keywords: IRF3; Src family kinase; TBK1; antiviral response; cytosolic nucleic acid sensing; innate immunity; kinase inhibitor; protein modification; tyrosine phosphorylation; zebrafish.

MeSH terms

  • Adaptor Proteins, Signal Transducing / metabolism
  • Animals
  • Antiviral Agents / immunology*
  • Antiviral Agents / metabolism
  • Cell Line
  • Chlorocebus aethiops
  • Cytosol / immunology
  • Cytosol / metabolism
  • HEK293 Cells
  • Hep G2 Cells
  • Herpesvirus 1, Human
  • Humans
  • Immunity, Innate*
  • Interferon Regulatory Factor-3 / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Phosphorylation
  • Protein Serine-Threonine Kinases / metabolism*
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-hck / metabolism
  • Respirovirus Infections / immunology
  • Rhabdoviridae Infections / immunology
  • Sendai virus / pathogenicity
  • Tyrosine / metabolism*
  • Ubiquitination
  • Vero Cells
  • Vesiculovirus
  • Virus Diseases / immunology*
  • Zebrafish / immunology
  • Zebrafish Proteins / metabolism
  • src-Family Kinases / antagonists & inhibitors*
  • src-Family Kinases / metabolism

Substances

  • Adaptor Proteins, Signal Transducing
  • Antiviral Agents
  • Interferon Regulatory Factor-3
  • Irf3 protein, mouse
  • Irf3 protein, zebrafish
  • Proto-Oncogene Proteins
  • Sh2d2a protein, mouse
  • Zebrafish Proteins
  • Tyrosine
  • Tbk1 protein, mouse
  • Hck protein, mouse
  • Proto-Oncogene Proteins c-hck
  • proto-oncogene proteins c-fgr
  • src-Family Kinases
  • Protein Serine-Threonine Kinases