[go: up one dir, main page]
More Web Proxy on the site http://driver.im/
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.

  • Letter
  • Published:

Distinct molecular mechanism for initiating TRAF6 signalling

Nature volume 418, pages 443–447 (2002)Cite this article

Abstract

Tumour-necrosis factor (TNF) receptor-associated factor 6 (TRAF6) is the only TRAF family member that participates in signal transduction of both the TNF receptor (TNFR) superfamily and the interleukin-1 receptor (IL-1R)/Toll-like receptor (TLR) superfamily1,2,3,4,5; it is important for adaptive immunity, innate immunity and bone homeostasis. Here we report crystal structures of TRAF6, alone and in complex with TRAF6-binding peptides from CD40 and TRANCE-R (also known as RANK), members of the TNFR superfamily, to gain insight into the mechanism by which TRAF6 mediates several signalling cascades. A 40° difference in the directions of the bound peptides in TRAF6 and TRAF2 shows that there are marked structural differences between receptor recognition by TRAF6 and other TRAFs. The structural determinant of the petide–TRAF6 interaction reveals a Pro-X-Glu-X-X-(aromatic/acidic residue) TRAF6-binding motif, which is present not only in CD40 and TRANCE-R but also in the three IRAK adapter kinases for IL-1R/TLR signalling. Cell-permeable peptides with the TRAF6-binding motif inhibit TRAF6 signalling, which indicates their potential as therapeutic modulators. Our studies identify a universal mechanism by which TRAF6 regulates several signalling cascades in adaptive immunity, innate immunity and bone homeostasis.

Access through your institution
Buy or subscribe

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

Access options

Access through your institution

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: TRAF6 structures.
Figure 2: Functional analyses for the interaction of TRAF6 with CD40, TRANCE-R and IRAK.
Figure 3: Inhibitory effects of TRAF6 decoy peptides (L-T6DP-1 and L-T6DP-2) in TRANCE-mediated signal transduction and osteoclast differentiation.

Similar content being viewed by others

References

  1. Cao, Z., Xiong, J., Takeuchi, M., Kurama, T. & Goeddel, D. V. TRAF6 is a signal transducer for interleukin-1. Nature 383, 443–446 (1996)

    Article  ADS  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Naito, A. et al. Severe osteopetrosis, defective interleukin-1 signalling and lymph node organogenesis in TRAF6-deficient mice. Genes Cells 4, 353–362 (1999)

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  5. Chung, J. Y., Park, Y. C., Ye, H. & Wu, H. All TRAFs are not created equal: common and distinct molecular mechanisms of TRAF-mediated signal transduction. J. Cell Sci. 115, 679–688 (2002)

    CAS  PubMed  Google Scholar 

  6. Park, Y. C., Burkitt, V., Villa, A. R., Tong, L. & Wu, H. Structural basis for self-association and receptor recognition of human TRAF2. Nature 398, 533–538 (1999)

    Article  ADS  CAS  PubMed  Google Scholar 

  7. McWhirter, S. M. et al. Crystallographic analysis of CD40 recognition and signalling by human TRAF2. Proc. Natl Acad. Sci. USA 96, 8408–8413 (1999)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ye, H., Park, Y. C., Kreishman, M., Kieff, E. & Wu, H. The structural basis for the recognition of diverse receptor sequences by TRAF2. Mol. Cell 4, 321–330 (1999)

    Article  CAS  PubMed  Google Scholar 

  9. Pullen, S. S. et al. CD40-tumour necrosis factor receptor-associated factor (TRAF) interactions: regulation of CD40 signalling through multiple TRAF binding sites and TRAF hetero-oligomerization. Biochemistry 37, 11836–11845 (1998)

    Article  CAS  PubMed  Google Scholar 

  10. Darnay, B. G., Ni, J., Moore, P. A. & Aggarwal, B. B. Activation of NF-κB by RANK requires tumour necrosis factor receptor-associated factor (TRAF) 6 and NF-κB-inducing kinase. Identification of a novel TRAF6 interaction motif. J. Biol. Chem. 274, 7724–7731 (1999)

    Article  CAS  PubMed  Google Scholar 

  11. Pullen, S. S., Dang, T. T., Crute, J. J. & Kehry, M. R. CD40 signalling through tumour necrosis factor receptor-associated factors (TRAFs). Binding site specificity and activation of downstream pathways by distinct TRAFs. J. Biol. Chem. 274, 14246–14254 (1999)

    Article  CAS  PubMed  Google Scholar 

  12. Wesche, H. et al. IRAK-M is a novel member of the Pelle/interleukin-1 receptor-associated kinase (IRAK) family. J. Biol. Chem. 274, 19403–19410 (1999)

    Article  CAS  PubMed  Google Scholar 

  13. Wong, B. R. et al. The TRAF family of signal transducers mediates NF-κB activation by the TRANCE receptor. J. Biol. Chem. 273, 28355–28359 (1998)

    Article  CAS  PubMed  Google Scholar 

  14. Cao, Z., Henzel, W. J. & Gao, X. IRAK: A kinase associated with the interleukin-1 receptor. Science 271, 1128–1131 (1996)

    Article  ADS  CAS  PubMed  Google Scholar 

  15. Zhang, F. X. et al. Bacterial lipopolysaccharide activates nuclear factor-κB through interleukin-1 signalling mediators in cultured human dermal endothelial cells and mononuclear phagocytes. J. Biol. Chem. 274, 7611–7614 (1999)

    Article  CAS  PubMed  Google Scholar 

  16. Muzio, M., Ni, J., Feng, P. & Dixit, V. M. IRAK (Pelle) family member IRAK-2 and MyD88 as proximal mediators of IL-1 signalling. Science 278, 1612–1615 (1997)

    Article  ADS  CAS  PubMed  Google Scholar 

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

    Article  ADS  CAS  PubMed  Google Scholar 

  18. Horng, T., Barton, G. M. & Medzhitov, R. TIRAP: an adapter molecule in the Toll signalling pathway. Nature Immunol. 2, 835–841 (2001)

    Article  CAS  Google Scholar 

  19. McCarthy, J. V., Ni, J. & Dixit, V. M. RIP2 is a novel NF-κB-activating and cell death-inducing kinase. J. Biol. Chem. 273, 16968–16975 (1998)

    Article  CAS  PubMed  Google Scholar 

  20. Yan Liu, X. et al. Peptide-directed suppression of a pro-inflammatory cytokine response. J. Biol. Chem. 275, 16774–16778 (2000)

    Article  Google Scholar 

  21. Arron, J. R. & Choi, Y. Bone versus immune system. Nature 408, 535–536 (2000)

    Article  ADS  CAS  PubMed  Google Scholar 

  22. Bundel, D. R. & Sigurskjold, B. W. Determination of accurate thermodynamics of binding by titration microcalorimetry. Methods Enymol. 247, 288–305 (1987)

    Article  Google Scholar 

  23. Otwinowski, Z. & Minor, W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276, 307–326 (1997)

    Article  CAS  PubMed  Google Scholar 

  24. Tong, L. REPLACE, a suite of computer programs for molecular-replacement calculations. J. Appl. Crystallogr. 26, 748–751 (1993)

    Article  Google Scholar 

  25. Brunger, A. T. et al. Crystallography & NMR system: A new software suite for macromolecular structure determination. Acta Crystallogr. D 54, 905–921 (1998)

    Article  CAS  PubMed  Google Scholar 

  26. Jones, T. A., Zou, J.-Y., Cowan, S. W. & Kjeldgaard, M. Improved methods for building models in electron density maps and the location of errors in those models. Acta Crystallgr. A 47, 110–119 (1991)

    Article  Google Scholar 

  27. Evans, S. V. SETOR: hardware-lighted three-dimensional solid model representations of macromolecules. J. Mol. Graph. 11, 134–138 (1993)

    Article  CAS  PubMed  Google Scholar 

  28. Nicholls, A., Sharp, K. A. & Honig, B. Protein folding and association: insights from the interfacial and thermodynamic properties of hydrocarbons. Proteins 11, 281–296 (1991)

    Article  CAS  PubMed  Google Scholar 

  29. Haridas, V., Darnay, B. G., Natarajan, K., Heller, R. & Aggarwal, B. B. Overexpression of the p80 TNF receptor leads to TNF-dependent apoptosis, nuclear factor-κB activation, and c-Jun kinase activation. J. Immunol. 160, 3152–3162 (1998)

    CAS  PubMed  Google Scholar 

  30. Shevde, N. K., Bendixen, A. C., Dienger, K. M. & Pike, J. W. Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression. Proc. Natl Acad. Sci. USA 97, 7829–7834 (2000)

    Article  ADS  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank the structural biology groups at the Memorial Sloan-Kettering Cancer Center for use of the microcalorimeter; Z. Cao for human TRAF6 cDNA; G. Mosialos and E. Kieff for the human GST–CD40ct construct; L. Tong, R. Khayat, Z. Yang and C. Lima for help with diffraction data collection; G. Cheng for discussions; C. Ogata and MacCHESS staff for beamline access and support; T. Burling for maintaining the home X-ray source; V. Burkitt and A. Villa for technical help; and laboratory members of Imgenex for synthesizing the decoy peptides. This work was supported in part by the NIH (Y.C.), an MSTP grant (J.R.A.), start-up funds from the Department of Bioimmunotherapy (B.G.D.) and a Translational Research Grant from the Leukemia and Lymphoma Society (B.G.D.). H.Y. is a postdoctoral fellow from the Revson Foundation. H.W. is a Pew Scholar of biomedical sciences and a Rita Allen Scholar.

Author information

Author notes

  1. Betty Lamothe

    Present address: Department of Pharmacology, Yale University, New Haven, Connecticut, 06520, USA

  2. Hong Ye, Joseph R. Arron and Masha Vologodskaia: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biochemistry, Weill Medical College of Cornell University, New York, New York, 10021, USA

    Hong Ye, Maurizio Cirilli, Deena Segal, Oki K. Dzivenu & Hao Wu

  2. Tri-Institutional MD-PhD Program, The Rockefeller University, New York, New York, 10021, USA

    Joseph R. Arron

  3. Laboratory of Immunology, The Rockefeller University, New York, New York, 10021, USA

    Masha Vologodskaia

  4. Abramson Family Cancer Research Institute, Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania, 19104, USA

    Joseph R. Arron, Takashi Kobayashi, Mijung Yim & Yongwon Choi

  5. Department of Bioimmunotherapy, The University of Texas MD Anderson Cancer Center, Houston, Texas, 77030, Box 143, 1515 Holcombe Boulevard, USA

    Betty Lamothe, Khoi Du & Bryant G. Darnay

  6. Istituto di Strutturistica Chimica ‘Giordano Giacomello’, CNR, CP 10, Italy, Monterotondo Stazione

    Maurizio Cirilli

  7. Department of Biochemistry, University of Wisconsin, Madison, Wisconsin, 53706, USA

    Nirupama K. Shevde & J. Wesley Pike

  8. Imgenex Corporation, San Diego, 11185 Flintkote Ave, Suite E, California, 92121, USA

    Sujay Singh

Authors
  1. Hong Ye
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joseph R. Arron
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Betty Lamothe
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maurizio Cirilli
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takashi Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Nirupama K. Shevde
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Deena Segal
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Oki K. Dzivenu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Masha Vologodskaia
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Mijung Yim
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Khoi Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Sujay Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. J. Wesley Pike
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Bryant G. Darnay
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Yongwon Choi
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. Hao Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hao Wu.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ye, H., Arron, J., Lamothe, B. et al. Distinct molecular mechanism for initiating TRAF6 signalling. Nature 418, 443–447 (2002). https://doi.org/10.1038/nature00888

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature00888

This article is cited by

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