RIP1 is an essential mediator of Toll-like receptor 3–induced NF-κB activation

Abstract

Stimulation of Toll-like receptors (TLRs) initiates potent innate immune responses through Toll–interleukin 1 receptor (TIR) domain–containing adaptors such as MyD88 and Trif. Analysis of Trif-deficient mice has shown that TLR3-dependent activation of the transcription factor NF-κB by the TLR3 ligand double-stranded RNA is Trif dependent. Here we investigated the 'downstream' signaling events that regulate TLR3-dependent Trif-induced NF-κB activation. Trif recruited the kinases receptor interacting protein (RIP)-1 and RIP3 through its RIP homotypic interaction motif. In the absence of RIP1, TLR3-mediated signals activating NF-κB, but not the kinase JNK or interferon-β, were abolished, suggesting that RIP1 mediates Trif-induced NF-κB activation. In contrast, the presence of RIP3 negatively regulated the Trif-RIP1–induced NF-κB pathway. Therefore, in contrast to other TLRs, which use interleukin 1 receptor-associated kinase (IRAK) proteins to activate NF-κB, TLR 3-induced NF-κB activation is dependent on RIP kinases.

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Figure 1: Trif interacts with RIP1 and RIP3.
Figure 2: Trif and RIP1 interact through their RHIM.
Figure 3: Trif-induced NF-κB activation is RHIM dependent.
Figure 4: RIP3 inhibits Trif-dependent activation of NF-κB.
Figure 5: TLR3-induced NF-κB activation is RIP1 dependent.

References

  1. 1

    Aderem, A. & Ulevitch, R.J. Toll-like receptors in the induction of the innate immune response. Nature 406, 782–787 (2000).

    CAS  Article  Google Scholar 

  2. 2

    Rock, F.L., Hardiman, G., Timans, J.C., Kastelein, R.A. & Bazan, J.F. A family of human receptors structurally related to Drosophila Toll. Proc. Natl. Acad. Sci. USA 95, 588–593 (1998).

    CAS  Article  Google Scholar 

  3. 3

    Akira, S., Takeda, K. & Kaisho, T. Toll-like receptors: critical proteins linking innate and acquired immunity. Nat. Immunol. 2, 675–680 (2001).

    CAS  Article  Google Scholar 

  4. 4

    O'Neill, L.A. & Dinarello, C.A. The IL-1 receptor/toll-like receptor superfamily: crucial receptors for inflammation and host defense. Immunol. Today 21, 206–209 (2000).

    CAS  Article  Google Scholar 

  5. 5

    Alexopoulou, L., Holt, A.C., Medzhitov, R. & Flavell, R.A. Recognition of double-stranded RNA and activation of NF-κB by Toll-like receptor 3. Nature 413, 732–738 (2001).

    CAS  Article  Google Scholar 

  6. 6

    Hoshino, K. et al. Cutting edge: Toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J. Immunol. 162, 3749–3752 (1999).

    CAS  PubMed  Google Scholar 

  7. 7

    Poltorak, A. et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 282, 2085–2088 (1998).

    CAS  Article  Google Scholar 

  8. 8

    Kawai, T. et al. Lipopolysaccharide stimulates the MyD88-independent pathway and results in activation of IFN-regulatory factor 3 and the expression of a subset of lipopolysaccharide-inducible genes. J. Immunol. 167, 5887–5894 (2001).

    CAS  Article  Google Scholar 

  9. 9

    Hoshino, K., Kaisho, T., Iwabe, T., Takeuchi, O. & Akira, S. Differential involvement of IFN-β in Toll-like receptor-stimulated dendritic cell activation. Int. Immunol. 14, 1225–1231 (2002).

    CAS  Article  Google Scholar 

  10. 10

    Oshiumi, H., Matsumoto, M., Funami, K., Akazawa, T. & Seya, T. TICAM-1, an adaptor molecule that participates in Toll-like receptor 3-mediated interferon-β induction. Nat. Immunol. 4, 161–167 (2003).

    CAS  Article  Google Scholar 

  11. 11

    Yamamoto, M. et al. Cutting edge: a novel Toll/IL-1 receptor domain-containing adapter that preferentially activates the IFN-β promoter in the Toll-like receptor signaling. J. Immunol. 169, 6668–6672 (2002).

    CAS  Article  Google Scholar 

  12. 12

    Yamamoto, M. et al. TRAM is specifically involved in the Toll-like receptor 4–mediated MyD88-independent signaling pathway. Nat. Immunol. 4, 1144–1150 (2003).

    CAS  Article  Google Scholar 

  13. 13

    Oshiumi, H. et al. TIR-containing adapter molecule (TICAM)-2, a bridging adapter recruiting to toll-like receptor 4 TICAM-1 that induces interferon-β. J. Biol. Chem. 278, 49751–49762 (2003).

    CAS  Article  Google Scholar 

  14. 14

    Fitzgerald, K.A. et al. IKKε and TBK1 are essential components of the IRF3 signaling pathway. Nat. Immunol. 4, 491–496 (2003).

    CAS  Article  Google Scholar 

  15. 15

    Sharma, S. et al. Triggering the interferon antiviral response through an IKK-related pathway. Science 300, 1148–1151 (2003).

    CAS  Article  Google Scholar 

  16. 16

    Sato, S. et al. Toll/IL-1 Receptor domain-containing adaptor inducing IFN-β (TRIF) associates with TNF receptor-associated factor 6 and TANK-binding kinase 1, and activates two distinct transcription factors, NF-κB and IFN-regulatory factor-3, in the Toll-like receptor signaling. J. Immunol. 171, 4304–4310 (2003).

    CAS  Article  Google Scholar 

  17. 17

    Hoebe, K. et al. Identification of Lps2 as a key transducer of MyD88-independent TIR signalling. Nature 424, 743–748 (2003).

    CAS  Article  Google Scholar 

  18. 18

    Yamamoto, M. et al. Role of adaptor TRIF in the MyD88-independent toll-like receptor signaling pathway. Science 301, 640–643 (2003).

    CAS  Article  Google Scholar 

  19. 19

    Lomaga, M.A. et al. TRAF6 deficiency results in osteopetrosis and defective interleukin-1, CD40, and LPS signaling. Genes Dev. 13, 1015–1024 (1999).

    CAS  Article  Google Scholar 

  20. 20

    Hsu, H., Huang, J., Shu, H.B., Baichwal, V. & Goeddel, D.V. TNF-dependent recruitment of the protein kinase RIP to the TNF receptor-1 signaling complex. Immunity 4, 387–396 (1996).

    CAS  Article  Google Scholar 

  21. 21

    Sun, X. et al. RIP3, a novel apoptosis-inducing kinase. J. Biol. Chem. 274, 16871–16875 (1999).

    CAS  Article  Google Scholar 

  22. 22

    Yu, P.W. et al. Identification of RIP3, a RIP-like kinase that activates apoptosis and NF-κB. Curr. Biol. 9, 539–542 (1999).

    CAS  Article  Google Scholar 

  23. 23

    Sun, X., Yin, J., Starovasnik, M.A., Fairbrother, W.J. & Dixit, V.M. Identification of a novel homotypic interaction motif required for the phosphorylation of receptor-interacting protein (RIP) by RIP3. J. Biol. Chem. 277, 9505–9511 (2002).

    CAS  Article  Google Scholar 

  24. 24

    Kelliher, M.A. et al. The death domain kinase RIP mediates the TNF-induced NF-κB signal. Immunity 8, 297–303 (1998).

    CAS  Article  Google Scholar 

  25. 25

    Xu, Y. et al. Structural basis for signal transduction by the Toll/interleukin-1 receptor domains. Nature 408, 111–115 (2000).

    CAS  Article  Google Scholar 

  26. 26

    Maniatis, T. et al. Structure and function of the interferon-β enhanceosome. Cold Spring Harb. Symp. Quant. Biol. 63, 609–620 (1998).

    CAS  Article  Google Scholar 

  27. 27

    Adachi, O. et al. Targeted disruption of the MyD88 gene results in loss of IL-1- and IL-18-mediated function. Immunity 9, 143–150 (1998).

    CAS  Article  Google Scholar 

  28. 28

    Meylan, E., Martinon, F., Thome, M., Gschwendt, M. & Tschopp, J. RIP4 (DIK/PKK), a novel member of the RIP kinase family, activates NF-κB and is processed during apoptosis. EMBO Rep. 3, 1201–1208 (2002).

    CAS  Article  Google Scholar 

  29. 29

    Janssens, S. & Beyaert, R. Functional diversity and regulation of different interleukin-1 receptor-associated kinase (IRAK) family members. Mol. Cell 11, 293–302 (2003).

    CAS  Article  Google Scholar 

  30. 30

    Kobayashi, K. et al. IRAK-M is a negative regulator of Toll-like receptor signaling. Cell 110, 191–202 (2002).

    CAS  Article  Google Scholar 

  31. 31

    Cusson, N., Oikemus, S., Kilpatrick, E.D., Cunningham, L. & Kelliher, M. The death domain kinase RIP protects thymocytes from tumor necrosis factor receptor type 2-induced cell death. J. Exp. Med. 196, 15–26 (2002).

    CAS  Article  Google Scholar 

  32. 32

    Thome, M. et al. Identification of CARDIAK, a RIP-like kinase that associates with caspase-1. Curr. Biol. 8, 885–888 (1998).

    CAS  Article  Google Scholar 

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Acknowledgements

We thank S. Hertig, N. Olivos, H. Everett, L. Agostini, M. Thome and B. Beutler for technical support, constructs and discussions. Supported by the Swiss National Science Foundation and Commission of Technology and Innovation.

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Correspondence to Jürg Tschopp.

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Meylan, E., Burns, K., Hofmann, K. et al. RIP1 is an essential mediator of Toll-like receptor 3–induced NF-κB activation. Nat Immunol 5, 503–507 (2004). https://doi.org/10.1038/ni1061

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