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Glycogen synthase kinase 3β ubiquitination by TRAF6 regulates TLR3-mediated pro-inflammatory cytokine production

Ryeojin Ko1, Jin Hee Park1, Hyunil Ha2

  • 1Department of Life Science and the Research Center for Cellular Homeostasis, Ewha Womans University, Seoul 120-750, South Korea.

Nature Communications
|April 2, 2015
PubMed

Insights

Glycogen synthase kinase 3 beta (GSK3β) positively regulates Toll-like receptor 3 (TLR3) signaling. TRAF6 ubiquitination of GSK3β is essential for inflammatory cytokine production via the TRIF complex.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Signaling

Background:

  • Toll-like receptor 3 (TLR3) pathways are crucial for innate immunity.
  • Tumor necrosis factor receptor-associated factor 6 (TRAF6) is vital for inflammatory cytokine production.
  • The precise role of TRAF6 in TLR3 signaling remains unclear.

Purpose of the Study:

  • To elucidate the mechanism by which TRAF6 regulates TLR3-mediated signaling.
  • To investigate the interaction between TRAF6 and Glycogen synthase kinase 3 beta (GSK3β) in TLR3 responses.

Main Methods:

  • Co-immunoprecipitation assays to detect protein interactions.
  • Western blotting to assess protein phosphorylation and ubiquitination.
  • Quantitative real-time PCR to measure cytokine production.
  • Gene silencing and kinase inhibition to study protein function.

Main Results:

  • GSK3β interacts with TRAF6 and enhances TLR3 signaling.
  • GSK3β suppression reduces inflammatory cytokine production and c-Fos induction by decreasing ERK and p38 phosphorylation.
  • TRAF6 directly ubiquitinates GSK3β in a poly I:C-dependent manner.
  • TRAF6-mediated GSK3β ubiquitination is critical for assembling the TRIF-containing signaling complex.

Conclusions:

  • GSK3β acts as a positive regulator of TLR3 signaling.
  • TRAF6-mediated ubiquitination of GSK3β is a key step in TLR3-induced inflammatory cytokine production.
  • This study reveals a novel regulatory mechanism in TLR3 innate immune responses.

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