Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

The zinc-finger protein ZFYVE1 modulates TLR3-mediated signaling by facilitating TLR3 ligand binding

Cellular & Molecular Immunology volume 17, pages 741–752 (2020)Cite this article

Subjects

Abstract

Recognition of viral dsRNA by Toll-like receptor 3 (TLR3) leads to the induction of downstream antiviral effectors and the innate antiviral immune response. Here, we identified the zinc-finger FYVE domain-containing protein ZFYVE1, a guanylate-binding protein (GBP), as a positive regulator of TLR3-mediated signaling. Overexpression of ZFYVE1 promoted the transcription of downstream antiviral genes upon stimulation with the synthetic TLR3 ligand poly(I:C). Conversely, ZFYVE1 deficiency had the opposite effect. Zfyve1−/− mice were less susceptible than wild-type mice to inflammatory death induced by poly(I:C) but not LPS. ZFYVE1 was associated with TLR3, and the FYVE domain of ZFYVE1 and the ectodomain of TLR3 were shown to be responsible for their interaction. ZFYVE1 was bound to poly(I:C) and increased the binding affinity of TLR3 to poly(I:C). These findings suggest that ZFYVE1 plays an important role in the TLR3-mediated innate immune and inflammatory responses by promoting the ligand binding of TLR3.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Kawai, T. & Akira, S. Toll-like receptors and their crosstalk with other innate receptors in infection and immunity. Immunity 34, 637–650 (2011).

    Article  CAS  Google Scholar 

  2. 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 

  3. Yang, Y. et al. The RNA-binding protein Mex3B is a coreceptor of Toll-like receptor 3 in innate antiviral response. Cell Res. 26, 288–303 (2016).

    Article  CAS  Google Scholar 

  4. Matsumoto, M. et al. Subcellular localization of Toll-like receptor 3 in human dendritic cells. J. Immunol. 171, 3154–3162 (2003).

    Article  CAS  Google Scholar 

  5. 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 

  6. Bell, J. K., Askins, J., Hall, P. R., Davies, D. R. & Segal, D. M. The dsRNA binding site of human Toll-like receptor 3. Proc. Natl. Acad. Sci. USA 103, 8792–8797 (2006).

    Article  CAS  Google Scholar 

  7. de Bouteiller, O. et al. Recognition of double-stranded RNA by human toll-like receptor 3 and downstream receptor signaling requires multimerization and an acidic pH. J. Biol. Chem. 280, 38133–38145 (2005).

    Article  Google Scholar 

  8. Wang, Y., Liu, L., Davies, D. R. & Segal, D. M. Dimerization of Toll-like receptor 3 (TLR3) is required for ligand binding. J. Biol. Chem. 285, 36836–36841 (2010).

    Article  CAS  Google Scholar 

  9. Yamashita, M. et al. Epidermal growth factor receptor is essential for Toll-like receptor 3 signaling. Sci. Signal. 5, ra50 (2012).

    Article  Google Scholar 

  10. 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 

  11. Qi, R., Singh, D. & Kao, C. C. Proteolytic processing regulates Toll-like receptor 3 stability and endosomal localization. J. Biol. Chem. 287, 32617–32629 (2012).

    Article  CAS  Google Scholar 

  12. Toscano, F. et al. Cleaved/associated TLR3 represents the primary form of the signaling receptor. J. Immunol. 190, 764–773 (2013).

    Article  CAS  Google Scholar 

  13. Zhu, S., Wang, G., Lei, X. & Flavell, R. A. Mex3B: a coreceptor to present dsRNA to TLR3. Cell Res. 26, 391–392 (2016).

    Article  Google Scholar 

  14. Johnsen, I. B. et al. Toll-like receptor 3 associates with c-Src tyrosine kinase on endosomes to initiate antiviral signaling. EMBO J. 25, 3335–3346 (2006).

    Article  CAS  Google Scholar 

  15. Hu, M. M. & Shu, H. B. Multifaceted roles of TRIM38 in innate immune and inflammatory responses. Cell Mol. Immunol. 14, 331–338 (2017).

    Article  CAS  Google Scholar 

  16. Luo, W. W. & Shu, H. B. Delicate regulation of the cGAS-MITA-mediated innate immune response. Cell Mol. Immunol. 15, 666–675 (2018).

    Article  CAS  Google Scholar 

  17. Hu, Y. H. et al. WDFY1 mediates TLR3/4 signaling by recruiting TRIF. EMBO Rep. 16, 447–455 (2015).

    Article  CAS  Google Scholar 

  18. Sanjana, N. E., Shalem, O. & Zhang, F. Improved vectors and genome-wide libraries for CRISPR screening. Nat. Methods 11, 783–784 (2014).

    Article  CAS  Google Scholar 

  19. Shalem, O. et al. Genome-scale CRISPR-Cas9 knockout screening in human cells. Science 343, 84–87 (2014).

    Article  CAS  Google Scholar 

  20. Ridley, S. H. et al. FENS-1 and DFCP1 are FYVE domain-containing proteins with distinct functions in the endosomal and Golgi compartments. J. Cell Sci. 114, 3991–4000 (2001).

    CAS  PubMed  Google Scholar 

  21. Nanao, T., Koike, M., Yamaguchi, J., Sasaki, M. & Uchiyama, Y. Cellular localization and tissue distribution of endogenous DFCP1 protein. Biomed. Res. 36, 121–133 (2015).

    Article  CAS  Google Scholar 

  22. He, J. et al. Membrane insertion of the FYVE domain is modulated by pH. Proteins 76, 852–860 (2009).

    Article  CAS  Google Scholar 

  23. Cheung, P. C., Trinkle-Mulcahy, L., Cohen, P. & Lucocq, J. M. Characterization of a novel phosphatidylinositol 3-phosphate-binding protein containing two FYVE fingers in tandem that is targeted to the Golgi. Biochem. J. 355, 113–121 (2001).

    Article  CAS  Google Scholar 

  24. Verma, R. & Bharti, K. Toll like receptor 3 and viral infections of nervous system. J. Neurol. Sci. 372, 40–48 (2017).

    Article  CAS  Google Scholar 

  25. Luo, W. W. et al. iRhom2 is essential for innate immunity to DNA viruses by mediating trafficking and stability of the adaptor STING. Nat. Immunol. 17, 1057–1066 (2016).

    Article  CAS  Google Scholar 

  26. Zhong, B. et al. The adaptor protein MITA links virus-sensing receptors to IRF3 transcription factor activation. Immunity 29, 538–550 (2008).

    Article  CAS  Google Scholar 

  27. Lian, H. et al. The zinc-finger protein ZCCHC3 binds RNA and facilitates viral RNA sensing and activation of the RIG-I-like receptors. Immunity 49, 438–448 (2018).

    Article  CAS  Google Scholar 

  28. Liu, Y. et al. ZDHHC11 modulates innate immune response to DNA virus by mediating MITA-IRF3 association. Cell. Mol. Immunol. 15, 907–916 (2018).

    Article  CAS  Google Scholar 

  29. Lin, H. et al. MARCH3 attenuates IL-1beta-triggered inflammation by mediating K48-linked polyubiquitination and degradation of IL-1RI. Proc. Natl. Acad. Sci. USA 115, 12483–12488 (2018).

    Article  CAS  Google Scholar 

  30. Jiang, L. Q. et al. IFITM3 inhibits virus-triggered induction of type I interferon by mediating autophagosome-dependent degradation of IRF3. Cell. Mol. Immunol. 15, 858–867 (2018).

    Article  CAS  Google Scholar 

  31. Hu, M. M. et al. Sumoylation promotes the stability of the DNA sensor cGAS and the adaptor STING to regulate the kinetics of response to DNA virus. Immunity 45, 555–569 (2016).

    Article  CAS  Google Scholar 

  32. Yang, Q. et al. TRIM32-TAX1BP1-dependent selective autophagic degradation of TRIF negatively regulates TLR3/4-mediated innate immune responses. PLoS Pathog. 13, e1006600 (2017).

    Article  Google Scholar 

  33. Lian, H. et al. ZCCHC3 is a co-sensor of cGAS for dsDNA recognition in innate immune response. Nat. Commun. 9, 3349 (2018).

    Article  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the State Key R&D Program of China (2017YFA0505800, 2016YFA0502102 and 2018YFA0800700), the National Natural Science Foundation of China (31830024, 31630045, 31870870, 31800728, 31771555, and 31671418), the China Postdoctoral Science Foundation (2017M620334), the Fundamental Research Funds for the Central Universities (2042019kf0204), and the Natural Science Foundation of Hubei Province (2018CFA016).

Author information

Authors and Affiliations

  1. College of Life Sciences, Wuhan University, Wuhan, 430072, China

    Xuan Zhong, Lu Feng, Wen-Hua Xu, Xin Wu, Yi-Di Ding, Yan Zhou, Cao-Qi Lei & Hong-Bing Shu

  2. Department of Infectious Diseases, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan, 430072, China

    Xuan Zhong, Lu Feng, Wen-Hua Xu, Xin Wu, Yan Zhou, Cao-Qi Lei & Hong-Bing Shu

  3. Medical Research Institute, Wuhan University, Wuhan, 430072, China

    Xuan Zhong, Lu Feng, Wen-Hua Xu, Xin Wu, Yan Zhou, Cao-Qi Lei & Hong-Bing Shu

Authors
  1. Xuan Zhong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lu Feng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wen-Hua Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Xin Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yi-Di Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yan Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Cao-Qi Lei
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Hong-Bing Shu
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

H.-B.S. and X.Z. designed the study; X.Z., L.F., W.-H.X., X.W., and Y.-D.D. performed the experiments; H.-B.S., C.-Q.L., Y.Z., and X.Z. analyzed the data; and H.-B.S., C.-Q.L., and X.Z. wrote the paper.

Corresponding authors

Correspondence to Cao-Qi Lei or Hong-Bing Shu.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhong, X., Feng, L., Xu, WH. et al. The zinc-finger protein ZFYVE1 modulates TLR3-mediated signaling by facilitating TLR3 ligand binding. Cell Mol Immunol 17, 741–752 (2020). https://doi.org/10.1038/s41423-019-0265-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41423-019-0265-6

Keywords

This article is cited by

Search

Advanced search

Quick links