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E3 ubiquitin ligase RNF170 inhibits innate immune responses by targeting and degrading TLR3 in murine cells

Cellular & Molecular Immunology volume 17, pages 865–874 (2020)Cite this article

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Abstract

Upon recognition of dsRNA, toll-like receptor 3 (TLR3) recruits the adaptor protein TRIF to activate IRF3 and NF-κB signaling, initiating innate immune responses. The ubiquitination of TLR3 downstream signaling molecules and their roles in the innate response have been discovered; however, whether TLR3 itself is ubiquitinated and then functionally involved remains to be elucidated. By immunoprecipitating TLR3-binding proteins in macrophages, we identified ring finger protein 170 (RNF170) as a TLR3-binding E3 ligase. RNF170 mediated the K48-linked polyubiquitination of K766 in the TIR domain of TLR3 and promoted the degradation of TLR3 through the proteasome pathway. The genetic ablation of RNF170 selectively augmented TLR3-triggered innate immune responses both in vitro and in vivo. Our results reveal a novel role for RNF170 in selectively inhibiting TLR3-triggered innate immune responses by promoting TLR3 degradation.

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References

  1. Kawai, T. & Akira, S. The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat. Immunol. 11, 373–384 (2010).

    Article  CAS  Google Scholar 

  2. Liu, L. et al. Structural basis of toll-like receptor 3 signaling with double-stranded RNA. Science 320, 379–381 (2008).

    Article  CAS  Google Scholar 

  3. McGettrick, A. F. & O’Neill, L. A. Localisation and trafficking of Toll-like receptors: an important mode of regulation. Curr. Opin. Immunol. 22, 20–27 (2010).

    Article  CAS  Google Scholar 

  4. Garcia-Cattaneo, A. et al. Cleavage of Toll-like receptor 3 by cathepsins B and H is essential for signaling. Proc. Natl Acad. Sci. USA 109, 9053–9058 (2012).

    Article  CAS  Google Scholar 

  5. Leonard, J. N. et al. The TLR3 signaling complex forms by cooperative receptor dimerization. Proc. Natl Acad. Sci. USA 105, 258–263 (2008).

    Article  CAS  Google Scholar 

  6. Roers, A., Hiller, B. & Hornung, V. Recognition of endogenous nucleic acids by the innate immune system. Immunity 44, 739–754 (2016).

    Article  CAS  Google Scholar 

  7. Daffis, S., Samuel, M. A., Suthar, M. S., Gale, M. & Diamond, M. S. Toll-Like receptor 3 has a protective role against West nile virus infection. J. Virol. 82, 10349–10358 (2008).

    Article  CAS  Google Scholar 

  8. Guillot, L. et al. Involvement of toll-like receptor 3 in the immune response of lung epithelial cells to double-stranded RNA and influenza A virus. J. Biol. Chem. 280, 5571–5580 (2005).

    Article  CAS  Google Scholar 

  9. Lafaille, F. G. et al. Impaired intrinsic immunity to HSV-1 in human iPSC-derived TLR3-deficient CNS cells. Nature 491, 769–773 (2012).

    Article  CAS  Google Scholar 

  10. Zhang, S. Y. et al. TLR3 deficiency in patients with herpes simplex encephalitis. Science 317, 1522–1527 (2007).

    Article  CAS  Google Scholar 

  11. Harii, N. et al. Thyrocytes express a functional toll-like receptor 3: overexpression can be induced by viral infection and reversed by phenylmethimazole and is associated with Hashimoto’s autoimmune thyroiditis. Mol. Endocrinol. 19, 1231–1250 (2005).

    Article  CAS  Google Scholar 

  12. Roelofs, M. F. et al. The expression of toll-like receptors 3 and 7 in rheumatoid arthritis synovium is increased and costimulation of toll-like receptors 3, 4, and 7/8 results in synergistic cytokine production by dendritic cells. Arthritis Rheum. 52, 2313–2322 (2005).

    Article  CAS  Google Scholar 

  13. Kondo, T., Kawai, T. & Akira, S. Dissecting negative regulation of Toll-like receptor signaling. Trends Immunol. 33, 449–458 (2012).

    Article  CAS  Google Scholar 

  14. Liu, J., Qian, C. & Cao, X. Post-translational modification control of innate immunity. Immunity 45, 15–30 (2016).

    Article  Google Scholar 

  15. Cao, X. Self-regulation and cross-regulation of pattern-recognition receptor signalling in health and disease. Nat. Rev. Immunol. 16, 35–50 (2016).

    Article  CAS  Google Scholar 

  16. Jiang, X. & Chen, Z. J. The role of ubiquitylation in immune defence and pathogen evasion. Nat. Rev. Immunol. 12, 35–48 (2011).

    Article  Google Scholar 

  17. Ko, R., Park, J. H., Ha, H., Choi, Y. & Lee, S. Y. Glycogen synthase kinase 3beta ubiquitination by TRAF6 regulates TLR3-mediated pro-inflammatory cytokine production. Nat. Commun. 6, 6765 (2015).

    Article  CAS  Google Scholar 

  18. Siednienko, J. et al. Pellino3 targets the IRF7 pathway and facilitates autoregulation of TLR3- and viral-induced expression of type I interferons. Nat. Immunol. 13, 1055–1062 (2012).

    Article  CAS  Google Scholar 

  19. Yang, Y. et al. E3 ligase WWP2 negatively regulates TLR3-mediated innate immune response by targeting TRIF for ubiquitination and degradation. Proc. Natl Acad. Sci. USA 110, 5115–5120 (2013).

    Article  CAS  Google Scholar 

  20. Lu, J. P., Wang, Y., Sliter, D. A., Pearce, M. M. & Wojcikiewicz, R. J. RNF170 protein, an endoplasmic reticulum membrane ubiquitin ligase, mediates inositol 1,4,5-trisphosphate receptor ubiquitination and degradation. J. Biol. Chem. 286, 24426–24433 (2011).

    Article  CAS  Google Scholar 

  21. Wang, W. et al. RNF122 suppresses antiviral type I interferon production by targeting RIG-I CARDs to mediate RIG-I degradation. Proc. Natl Acad. Sci. USA 113, 9581–9586 (2016).

    Article  CAS  Google Scholar 

  22. Wright, F. A. et al. A point mutation in the ubiquitin ligase RNF170 that causes autosomal dominant sensory ataxia destabilizes the protein and impairs inositol 1,4,5-trisphosphate receptor-mediated Ca2+signaling. J. Biol. Chem. 290, 13948–13957 (2015).

    Article  CAS  Google Scholar 

  23. Chuang, T. H. & Ulevitch, R. J. Triad3A, an E3 ubiquitin-protein ligase regulating Toll-like receptors. Nat. Immunol. 5, 495–502 (2004).

    Article  CAS  Google Scholar 

  24. Kumazoe, M. et al. Green Tea polyphenol epigallocatechin-3-gallate suppresses toll-like receptor 4 expression via up-regulation of E3 ubiquitin-protein ligase RNF216. J. Biol. Chem. 292, 4077–4088 (2017).

    Article  CAS  Google Scholar 

  25. McKelvey, A. C. et al. RING finger E3 ligase PPP1R11 regulates TLR2 signaling and innate immunity. Elife 5, e18496 (2016).

    Article  Google Scholar 

  26. Wang, Y. et al. Lysosome-associated small Rab GTPase Rab7b negatively regulates TLR4 signaling in macrophages by promoting lysosomal degradation of TLR4. Blood 110, 962–971 (2007).

    Article  CAS  Google Scholar 

  27. Blasius, A. L. & Beutler, B. Intracellular toll-like receptors. Immunity 32, 305–315 (2010).

    Article  CAS  Google Scholar 

  28. Schroder, M. & Bowie, A. G. TLR3 in antiviral immunity: key player or bystander? Trends Immunol. 26, 462–468 (2005).

    Article  Google Scholar 

  29. Schulz, O. et al. Toll-like receptor 3 promotes cross-priming to virus-infected cells. Nature 433, 887–892 (2005).

    Article  CAS  Google Scholar 

  30. Cario, E. & Podolsky, D. K. Differential alteration in intestinal epithelial cell expression of toll-like receptor 3 (TLR3) and TLR4 in inflammatory bowel disease. Infect. Immun. 68, 7010–7017 (2000).

    Article  CAS  Google Scholar 

  31. Tilton, J. C. et al. Diminished production of monocyte proinflammatory cytokines during human immunodeficiency virus viremia is mediated by type I interferons. J. Virol. 80, 11486–11497 (2006).

    Article  CAS  Google Scholar 

  32. Hertel, L. & Mocarski, E. S. Global analysis of host cell gene expression late during cytomegalovirus infection reveals extensive dysregulation of cell cycle gene expression and induction of Pseudomitosis independent of US28 function. J. Virol. 78, 11988–12011 (2004).

    Article  CAS  Google Scholar 

  33. Netea, M. G., Wijmenga, C. & O’Neill, L. A. Genetic variation in Toll-like receptors and disease susceptibility. Nat. Immunol. 13, 535–542 (2012).

    Article  CAS  Google Scholar 

  34. Tabeta, K. et al. The Unc93b1 mutation 3d disrupts exogenous antigen presentation and signaling via Toll-like receptors 3, 7 and 9. Nat. Immunol. 7, 156–164 (2006).

    Article  CAS  Google Scholar 

  35. Brinkmann, M. M. et al. The interaction between the ER membrane protein UNC93B and TLR3, 7, and 9 is crucial for TLR signaling. J. Cell Biol. 177, 265–275 (2007).

    Article  CAS  Google Scholar 

  36. Kim, Y. M., Brinkmann, M. M., Paquet, M. E. & Ploegh, H. L. UNC93B1 delivers nucleotide-sensing toll-like receptors to endolysosomes. Nature 452, 234–238 (2008).

    Article  CAS  Google Scholar 

  37. Kim, Y. et al. Age-dependent gait abnormalities in mice lacking the Rnf170 gene linked to human autosomal-dominant sensory ataxia. Hum. Mol. Genet. 24, 7196–7206 (2015).

    Article  CAS  Google Scholar 

  38. Valdmanis, P. N. et al. A mutation in the RNF170 gene causes autosomal dominant sensory ataxia. Brain 134, 602–607 (2011).

    Article  Google Scholar 

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (grant 81788101 to X.C.; grant 81871236 to M.J.), the National 135 Mega Program of China (grant 2017ZX10202203-002 to M.J.; grant 2017ZX10203206-001 to X.C.), the CAMS Innovation Fund for Medical Sciences (grant 2016-12M-1-003 to X.C.) and the Medical Epigenetics Research Center, Chinese Academy of Medical Sciences (2018PT31015).

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Authors and Affiliations

  1. National Key Laboratory of Medical Molecular Biology, Department of Immunology & Center for Immunotherapy, Institute of Basic Medical Sciences Chinese Academy of Medical Sciences, School of Basic Medicine Peking Union Medical College, 100005, Beijing, China

    Xiaoqi Song, Shuo Liu, Wendie Wang, Zhongfei Ma, Xuetao Cao & Minghong Jiang

  2. National Key Laboratory of Medical Immunology & Institute of Immunology, Second Military Medical University, 200433, Shanghai, China

    Xuetao Cao

  3. School of Medicine, Nankai University, 300071, Tianjin, China

    Xuetao Cao

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  1. Xiaoqi Song
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  2. Shuo Liu
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  3. Wendie Wang
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  4. Zhongfei Ma
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Contributions

M.J. and X.C. designed the research; X.S., M.J., S.L., W.W. and Z.M. performed the experiments; and X.S., M.J. and X.C. analyzed the data and wrote the paper.

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Correspondence to Minghong Jiang.

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Song, X., Liu, S., Wang, W. et al. E3 ubiquitin ligase RNF170 inhibits innate immune responses by targeting and degrading TLR3 in murine cells. Cell Mol Immunol 17, 865–874 (2020). https://doi.org/10.1038/s41423-019-0236-y

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