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:

Suppressor of cytokine signaling 1 negatively regulates Toll-like receptor signaling by mediating Mal degradation

Nature Immunology volume 7, pages 148–155 (2006)Cite this article

Abstract

Toll-like receptor (TLR) signals that initiate innate immune responses to pathogens must be tightly regulated to prevent excessive inflammatory damage to the host. The adaptor protein Mal is specifically involved in signaling via TLR2 and TLR4. We demonstrate here that after TLR2 and TLR4 stimulation Mal becomes phosphorylated by Bruton's tyrosine kinase (Btk) and then interacts with SOCS-1, which results in Mal polyubiquitination and subsequent degradation. Removal of SOCS-1 regulation potentiates Mal-dependent p65 phosphorylation and transactivation of NF-κB, leading to amplified inflammatory responses. These data identify a target of SOCS-1 that regulates TLR signaling via a mechanism distinct from an autocrine cytokine response. The transient activation of Mal and subsequent SOCS-1–mediated degradation is a rapid and selective means of limiting primary innate immune response.

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

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

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

Figure 1: Mal contains a PEST domain which targets it for TLR-mediated degradation.
Figure 2: Mal interacts with SOCS-1 and undergoes SOCS-1–mediated degradation.
Figure 3: SOCS-1 specifically promotes polyubiquitination of Mal.
Figure 4: Tyrosine phosphorylation by Btk is required to facilitate TLR-induced degradation of Mal.
Figure 5: Mal undergoes polyubiquitination on Lys 15 and 16 by SOCS-1, the dysregulation of which induces a potentiated NF-κB response.
Figure 6: SOCS-1 negative regulation of Mal controls the transient transactivation of NF-κB.

Similar content being viewed by others

References

  1. Takeda, K., Kaisho, T. & Akira, S. Toll-like receptors. Annu. Rev. Immunol. Vol. 21 (2003).

  2. Dunne, A. & O'Neill, L.A. Adaptor usage and Toll-like receptor signaling specificity. FEBS Lett. 579, 3330–3335 (2005).

    Article  CAS  Google Scholar 

  3. Akira, S. & Takeda, K. Toll-like receptor signalling. Nat. Rev. Immunol. 4, 499–511 (2004).

    Article  CAS  Google Scholar 

  4. O'Neill, L.A., Fitzgerald, K.A. & Bowie, A.G. The Toll-IL-1 receptor adaptor family grows to five members. Trends Immunol. 24, 286–290 (2003).

    Article  Google Scholar 

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

    Article  CAS  Google Scholar 

  6. Fitzgerald, K.A. et al. LPS-TLR4 signaling to IRF-3/7 and NF-kappaB involves the toll adapters TRAM and TRIF. J. Exp. Med. 198, 1043–1055 (2003).

    Article  CAS  Google Scholar 

  7. Horng, T., Barton, G.M., Flavell, R.A. & Medzhitov, R. The adaptor molecule TIRAP provides signalling specificity for Toll-like receptors. Nature 420, 329–333 (2002).

    Article  CAS  Google Scholar 

  8. Yamamoto, M. et al. Essential role for TIRAP in activation of the signalling cascade shared by TLR2 and TLR4. Nature 420, 324–329 (2002).

    Article  CAS  Google Scholar 

  9. Mansell, A., Brint, E., Gould, J.A., O'Neill, L.A. & Hertzog, P.J. Mal interacts with TNF receptor associated factor (TRAF)-6 to mediate NF-κB activation by Toll-like receptor (TLR)-2 and TLR4. J. Biol. Chem. 279, 37227–37230 (2004).

    Article  CAS  Google Scholar 

  10. Liew, F.Y., Xu, D., Brint, E.K. & O'Neill, L.A. Negative regulation of toll-like receptor-mediated immune responses. Nat. Rev. Immunol. 5, 446–458 (2005).

    Article  CAS  Google Scholar 

  11. 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 

  12. Wald, D. et al. SIGIRR, a negative regulator of Toll-like receptor-interleukin 1 receptor signaling. Nat. Immunol. 4, 920–927 (2003).

    Article  CAS  Google Scholar 

  13. Janssens, S., Burns, K., Vercammen, E., Tschopp, J. & Beyaert, R. MyD88S, a splice variant of MyD88, differentially modulates NF-kappaB- and AP-1-dependent gene expression. FEBS Lett. 548, 103–107 (2003).

    Article  CAS  Google Scholar 

  14. Naiki, Y. et al. Transforming growth factor-beta differentially inhibits MyD88-dependent, but not TRAM- and TRIF-dependent, lipopolysaccharide-induced TLR4 signaling. J. Biol. Chem. 280, 5491–5495 (2005).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  16. Divanovic, S. et al. Negative regulation of Toll-like receptor 4 signaling by the Toll-like receptor homolog RP105. Nat. Immunol. 6, 571–578 (2005).

    Article  CAS  Google Scholar 

  17. Naka, T. et al. Structure and function of a new STAT-induced STAT inhibitor. Nature 387, 924–929 (1997).

    Article  CAS  Google Scholar 

  18. Endo, T.A. et al. A new protein containing an SH2 domain that inhibits JAK kinases. Nature 387, 921–924 (1997).

    Article  CAS  Google Scholar 

  19. Starr, R. et al. A family of cytokine-inducible inhibitors of signalling. Nature 387, 917–921 (1997).

    Article  CAS  Google Scholar 

  20. Alexander, W.S. & Hilton, D.J. The role of suppressors of cytokine signaling (SOCS) proteins in regulation of the immune response. Annu. Rev. Immunol. 22, 503–529 (2004).

    Article  CAS  Google Scholar 

  21. Alexander, W.S. et al. SOCS1 is a critical inhibitor of interferon gamma signaling and prevents the potentially fatal neonatal actions of this cytokine. Cell 98, 597–608 (1999).

    Article  CAS  Google Scholar 

  22. Baetz, A., Frey, M., Heeg, K. & Dalpke, A.H. Suppressor of cytokine signaling (SOCS) proteins indirectly regulate toll-like receptor signaling in innate immune cells. J. Biol. Chem. 279, 54708–54715 (2004).

    Article  CAS  Google Scholar 

  23. Kinjyo, I. et al. SOCS1/JAB is a negative regulator of LPS-induced macrophage activation. Immunity 17, 583–591 (2002).

    Article  CAS  Google Scholar 

  24. Nakagawa, R. et al. SOCS-1 participates in negative regulation of LPS responses. Immunity 17, 677–687 (2002).

    Article  CAS  Google Scholar 

  25. Kamizono, S. et al. The SOCS box of SOCS-1 accelerates ubiquitin-dependent proteolysis of TEL-JAK2. J. Biol. Chem. 276, 12530–12538 (2001).

    Article  CAS  Google Scholar 

  26. Zhang, J.G. et al. The conserved SOCS box motif in suppressors of cytokine signaling binds to elongins B and C and may couple bound proteins to proteasomal degradation. Proc. Natl. Acad. Sci. USA 96, 2071–2076 (1999).

    Article  CAS  Google Scholar 

  27. Rechsteiner, M. & Rogers, S.W. PEST sequences and regulation by proteolysis. Trends Biochem. Sci. 21, 267–271 (1996).

    Article  CAS  Google Scholar 

  28. Kile, B.T. et al. The SOCS box: a tale of destruction and degradation. Trends Biochem. Sci. 27, 235–241 (2002).

    Article  CAS  Google Scholar 

  29. Nicholson, S.E. et al. Mutational analyses of the SOCS proteins suggest a dual domain requirement but distinct mechanisms for inhibition of LIF and IL-6 signal transduction. EMBO J. 18, 375–385 (1999).

    Article  CAS  Google Scholar 

  30. Rawlings, D.J. et al. Mutation of unique region of Bruton's tyrosine kinase in immunodeficient XID mice. Science 261, 358–361 (1993).

    Article  CAS  Google Scholar 

  31. Glickman, M.H. & Ciechanover, A. The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. Physiol. Rev. 82, 373–428 (2002).

    Article  CAS  Google Scholar 

  32. Fitzgerald, K.A. et al. Mal (MyD88-adapter-like) is required for Toll-like receptor-4 signal transduction. Nature 413, 78–83 (2001).

    Article  CAS  Google Scholar 

  33. Vermeulen, L., De Wilde, G., Notebaert, S., Vanden Berghe, W. & Haegeman, G. Regulation of the transcriptional activity of the nuclear factor-kappaB p65 subunit. Biochem. Pharmacol. 64, 963–970 (2002).

    Article  CAS  Google Scholar 

  34. Doyle, S.L., Jefferies, C.A. & O'Neill, L.A. Bruton's tyrosine kinase is involved in p65-mediated transactivation and phosphorylation of p65 on serine 536 during NFkappaB activation by lipopolysaccharide. J. Biol. Chem. 280, 23496–23501 (2005).

    Article  CAS  Google Scholar 

  35. Gingras, S., Parganas, E., de Pauw, A., Ihle, J.N. & Murray, P.J. Re-examination of the role of suppressor of cytokine signaling 1 (SOCS1) in the regulation of toll-like receptor signaling. J. Biol. Chem. 279, 54702–54707 (2004).

    Article  CAS  Google Scholar 

  36. Inagaki-Ohara, K., Hanada, T. & Yoshimura, A. Negative regulation of cytokine signaling and inflammatory diseases. Curr. Opin. Pharmacol. 3, 435–442 (2003).

    Article  CAS  Google Scholar 

  37. Yoshimura, A., Ohishi, H.M., Aki, D. & Hanada, T. Regulation of TLR signaling and inflammation by SOCS family proteins. J. Leukoc. Biol. 75, 422–427 (2004).

    Article  CAS  Google Scholar 

  38. Naka, T., Fujimoto, M. & Kishimoto, T. Negative regulation of cytokine signaling: STAT-induced STAT inhibitor. Trends Biochem. Sci. 24, 394–398 (1999).

    Article  CAS  Google Scholar 

  39. Fenner, J.E. et al. Suppressor of cytokine signaling 1 regulates the immune response to infection by a unique inhibition of type I interferon activity. Nat. Immunol. 7, 33–39 (2005).

    Article  Google Scholar 

  40. Noubir, S., Hmama, Z. & Reiner, N.E. Dual receptors and distinct pathways mediate interleukin-1 receptor-associated kinase degradation in response to lipopolysaccharide. Involvement of CD14/TLR4, CR3, and phosphatidylinositol 3-kinase. J. Biol. Chem. 279, 25189–25195 (2004).

    Article  CAS  Google Scholar 

  41. Yamin, T.T. & Miller, D.K. The interleukin-1 receptor-associated kinase is degraded by proteasomes following its phosphorylation. J. Biol. Chem. 272, 21540–21547 (1997).

    Article  CAS  Google Scholar 

  42. Sweet, M.J. et al. A novel pathway regulating lipopolysaccharide-induced shock by ST2/T1 via inhibition of Toll-like receptor 4 expression. J. Immunol. 166, 6633–6639 (2001).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Lang, J. Kiu and K. Palmer for technical assistance. Supported by the National Health and Medical Research Council (334023 to A.M., 236866 to P.C. and 284220 to P.J.H.), Science Foundation Ireland and the Cooperative Research Centre for Oral Health Sciences (R.S.).

Author information

Author notes

  1. Luke A J O'Neill and Paul J Hertzog: These authors contributed equally to this work.

Authors and Affiliations

  1. Centre for Functional Genomics and Human Disease, Monash Institute of Medical Research, Monash University, Melbourne, Victoria, Australia

    Ashley Mansell, Rosealee Smith, Jennifer E Fenner, Peter J Crack & Paul J Hertzog

  2. School of Biochemistry and Immunology, Trinity College, Dublin, 2, Ireland

    Sarah L Doyle, Pearl Gray & Luke A J O'Neill

  3. Walter and Eliza Hall Institute, Parkville, Victoria, Australia

    Sandra E Nicholson & Douglas J Hilton

Authors
  1. Ashley Mansell
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rosealee Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sarah L Doyle
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Pearl Gray
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jennifer E Fenner
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Peter J Crack
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sandra E Nicholson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Douglas J Hilton
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Luke A J O'Neill
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Paul J Hertzog
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Paul J Hertzog.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Schematic diagram illustrating the location within the N-terminal of the proline, glutamic acid, serine and threonine (PEST) rich domain located between residues human Mal (36-72) and murine Mal (43-76), proximal to their respective TIR domains; hMal (75-235), mMal (94-241). (PDF 125 kb)

Supplementary Fig. 2

Schematic diagram illustrating a role for Mal in mediating p65Ser536 phosphorylation of NF-κB, thereby inducing transcriptional activation of NF-κB, and its negative regulation by SOCS-1. (PDF 60 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mansell, A., Smith, R., Doyle, S. et al. Suppressor of cytokine signaling 1 negatively regulates Toll-like receptor signaling by mediating Mal degradation. Nat Immunol 7, 148–155 (2006). https://doi.org/10.1038/ni1299

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni1299

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing