Tankyrase represses autoinflammation through the attenuation of TLR2 signaling
Yoshinori Matsumoto, … , Jun Wada, Robert Rottapel
Yoshinori Matsumoto, … , Jun Wada, Robert Rottapel
Published April 1, 2022
Citation Information: J Clin Invest. 2022;132(7):e140869. https://doi.org/10.1172/JCI140869.
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Research Article Autoimmunity Inflammation

Tankyrase represses autoinflammation through the attenuation of TLR2 signaling

Abstract

Dysregulation of Toll-like receptor (TLR) signaling contributes to the pathogenesis of autoimmune diseases. Here, we provide genetic evidence that tankyrase, a member of the poly(ADP-ribose) polymerase (PARP) family, negatively regulates TLR2 signaling. We show that mice lacking tankyrase in myeloid cells developed severe systemic inflammation with high serum inflammatory cytokine levels. We provide mechanistic evidence that tankyrase deficiency resulted in tyrosine phosphorylation and activation of TLR2 and show that phosphorylation of tyrosine 647 within the TIR domain by SRC and SYK kinases was critical for TLR2 stabilization and signaling. Last, we show that the elevated cytokine production and inflammation observed in mice lacking tankyrase in myeloid cells were dependent on the adaptor protein 3BP2, which is required for SRC and SYK activation. These data demonstrate that tankyrase provides a checkpoint on the TLR-mediated innate immune response.

Authors

Yoshinori Matsumoto, Ioannis D. Dimitriou, Jose La Rose, Melissa Lim, Susan Camilleri, Napoleon Law, Hibret A. Adissu, Jiefei Tong, Michael F. Moran, Andrzej Chruscinski, Fang He, Yosuke Asano, Takayuki Katsuyama, Ken-ei Sada, Jun Wada, Robert Rottapel

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Figure 8

Tyrosine phosphorylation of TLR2 regulates NF-κB–mediated cytokine production.

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Tyrosine phosphorylation of TLR2 regulates NF-κB–mediated cytokine produ...
(A and B) Whole-cell lysates from primary murine macrophages derived from Tnks+/+ Tnks2fl/fl (WT) and Tnks–/– Tnks2fl/fl LysM-Cre (KO) mice, starved for 12 hours with 0.1% FBS and cultured in the presence of HKSA (107 cells/mL) for 0–30 minutes, were probed with the indicated antibodies for Western blot analysis. (C and G) Primary murine macrophages derived from Tnks+/+ Tnks2fl/fl and Tnks–/– Tnks2fl/fl LysM-Cre mice were starved for 12 hours with 0.1% FBS and cultured in the presence of HKSA (107 cells/mL) for 20 minutes, and SYK (C) or MyD88 (G) immune complexes were probed with the indicated antibodies for Western blot analysis. (D) Primary murine macrophages derived from Tnks+/+ Tnks2fl/fl and Tnks–/– Tnks2fl/fl LysM-Cre mice were starved for 12 hours with 0.1% FBS and cultured in the presence of HKSA (107 cells/mL) for 0–30 minutes, and pY immune complexes were probed with the indicated antibodies for Western blot analysis. (E) Whole-cell lysates from primary murine macrophages derived from Tnks+/+ Tnks2fl/fl and Tnks–/– Tnks2fl/fl LysM-Cre mice were probed with the indicated antibodies for Western blot analysis. (F) Nonradioactive pulse-chase assay to analyze turnover of endogenous TLR2 protein labeled with Click-IT Metabolic Labeling in primary murine macrophages derived from Tnks+/+ Tnks2fl/fl and Tnks–/– Tnks2fl/fl LysM-Cre mice. Biotinylated proteins were probed with the indicated antibodies, and percentages of TLR2 protein levels were plotted as a function of time. (H) qPCR analysis of Tnfa mRNA expression in primary murine macrophages derived from Tnks+/+ Tnks2fl/fl (WT) and Tnks–/– Tnks2fl/fl LysM-Cre (KO) mice and cultured in the presence or absence of SYK inhibitor (10–50 μM) and PP2 (10 μM); n = 3. P values were determined by unpaired t test (C, E, and G) or ANOVA with Tukey-Kramer post hoc test (A, B, D, and H). Data are presented as mean ± SEM. *P < 0.05.