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 1

TIR domain tyrosines are required for optimal TLR2 signaling.

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TIR domain tyrosines are required for optimal TLR2 signaling. (A) Primar...
(A) Primary murine macrophages derived from WT mice were starved for 12 hours with 0.1% FBS and cultured in the presence or absence of HKSA (107 cells/mL) for 20 minutes, and 3BP2 immune complexes were probed with the indicated antibodies for Western blot analysis. (B) Primary murine macrophages derived from Sh3bp2+/+ and Sh3bp2–/– mice were starved for 12 hours with 0.1% FBS and cultured in the presence of HKSA (107 cells/mL) for 20 minutes, and MyD88 immune complexes were probed with the indicated antibodies for Western blot analysis. (C) Luciferase activity from an NF-κB reporter assay in HEK293T cells cotransfected with the indicated constructs; n = 3. (D) Three tandem mass spectra of SRC-unique peptides in a TLR2 AP-MS sample. The y and b series of ions are indicated. The sequence of the peptide derived from spectra is shown at the top of each panel. (E and F) HEK293T cells were cotransfected with the indicated constructs, and HA-SRC (E) or HA-SYK (F) immune complexes were probed with the indicated antibodies for Western blot analysis. (G and H) Luciferase activity from an NF-κB reporter assay in HEK293T cells cotransfected with the indicated constructs; n = 3. (I) Luciferase activity from an NF-κB reporter assay in HEK293T cells cotransfected with the indicated constructs and cultured in the presence or absence of SYK inhibitor (10 μM) and PP2 (10 μM); n = 3. (J–L) HEK293T cells were cotransfected with the indicated constructs, and Myc-TLR2 (J), HA-SYK (K), or Flag-TLR2 (L) immune complexes were probed with the indicated antibodies for Western blot analysis. P values were determined by unpaired t test (A, B, K, and L) or ANOVA with Tukey-Kramer post hoc test (C and G–J). Data are presented as mean ± SEM. *P < 0.05.