Tyrosine phosphorylation of IRF3 by BLK facilitates its sufficient activation and innate antiviral response

PLoS Pathog. 2023 Oct 23;19(10):e1011742. doi: 10.1371/journal.ppat.1011742. eCollection 2023 Oct.

Abstract

Viral infection triggers the activation of transcription factor IRF3, and its activity is precisely regulated for robust antiviral immune response and effective pathogen clearance. However, how full activation of IRF3 is achieved has not been well defined. Herein, we identified BLK as a key kinase that positively modulates IRF3-dependent signaling cascades and executes a pre-eminent antiviral effect. BLK deficiency attenuates RNA or DNA virus-induced ISRE activation, interferon production and the cellular antiviral response in human and murine cells, whereas overexpression of BLK has the opposite effects. BLK-deficient mice exhibit lower serum cytokine levels and higher lethality after VSV infection. Moreover, BLK deficiency impairs the secretion of downstream antiviral cytokines and promotes Senecavirus A (SVA) proliferation, thereby supporting SVA-induced oncolysis in an in vivo xenograft tumor model. Mechanistically, viral infection triggers BLK autophosphorylation at tyrosine 309. Subsequently, activated BLK directly binds and phosphorylates IRF3 at tyrosine 107, which further promotes TBK1-induced IRF3 S386 and S396 phosphorylation, facilitating sufficient IRF3 activation and downstream antiviral response. Collectively, our findings suggest that targeting BLK enhances viral clearance via specifically regulating IRF3 phosphorylation by a previously undefined mechanism.

MeSH terms

  • Animals
  • Cytokines / metabolism
  • Humans
  • Immunity, Innate
  • Interferon Regulatory Factor-3 / metabolism
  • Mice
  • Phosphorylation
  • Protein Processing, Post-Translational
  • Protein Serine-Threonine Kinases* / metabolism
  • Signal Transduction
  • Virus Diseases*
  • src-Family Kinases / metabolism

Substances

  • Protein Serine-Threonine Kinases
  • Interferon Regulatory Factor-3
  • Cytokines
  • IRF3 protein, human
  • BLK protein, human
  • src-Family Kinases

Grants and funding

This work was supported by the grants from the National Key R&D Program of China (2021YFD1801300, awarded to H.X.Z.), the National Natural Science Foundation of China (32202781, awarded to W.W.L.), the Fundamental Research Funds for the Central Universities (awarded to H.X.Z.), the Key Technologies R&D Program of Gansu Province (21ZD3NA001, awarded to H.X.Z., 20ZD7NA006, awarded to H.X.Z., and 19ZD2NA001, awarded to H.X.Z.), the Natural Science Foundation of Gansu Province (22JR5RA034, awarded to W.W.L.), the Open Competition Program of Top Ten Critical Priorities of Agricultural Science and Technology Innovation for the 14th Five-Year Plan of Guangdong Province (2023SDZG02, awarded to H.X.Z.), the Earmarked Fund for CARS-35 (awarded to H.X.Z.) and CARS-39-13 (awarded to H.X.Z.), the Chinese Academy of Agricultural Science and Technology Innovation Project (CAAS-ZDRW202006, awarded to H.X.Z., and CAAS-ASTIP-2022-LVRI, awarded to H.X.Z.), and the Basic Scientific Research Fund of LVRI (1610312021009, awarded to W.W.L.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.