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:

Nonredundant and complementary functions of TRAF2 and TRAF3 in a ubiquitination cascade that activates NIK-dependent alternative NF-κB signaling

Abstract

The adaptor and signaling proteins TRAF2, TRAF3, cIAP1 and cIAP2 may inhibit alternative nuclear factor-κB (NF-κB) signaling in resting cells by targeting NF-κB–inducing kinase (NIK) for ubiquitin-dependent degradation, thus preventing processing of the NF-κB2 precursor protein p100 to release p52. However, the respective functions of TRAF2 and TRAF3 in NIK degradation and activation of alternative NF-κB signaling have remained elusive. We now show that CD40 or BAFF receptor activation result in TRAF3 degradation in a cIAP1-cIAP2- and TRAF2-dependent way owing to enhanced cIAP1, cIAP2 TRAF3-directed ubiquitin ligase activity. Receptor-induced activation of cIAP1 and cIAP2 correlated with their K63-linked ubiquitination by TRAF2. Degradation of TRAF3 prevented association of NIK with the cIAP1-cIAP2-TRAF2 ubiquitin ligase complex, which resulted in NIK stabilization and NF-κB2-p100 processing. Constitutive activation of this pathway causes perinatal lethality and lymphoid defects.

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

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

Figure 1: Deletion of NIK prevents postnatal lethality and splenic atrophy in Traf2−/− and Traf3−/− mice.
Figure 2: Reversal of lymphoid abnormalities in Traf2−/− and Traf3−/− mice by deletion of NIK.
Figure 3: TRAF2 and TRAF3 are involved in NIK turnover and NF-κB2 p100 processing.
Figure 4: TRAF3 links TRAF2 and cIAP2 to NIK.
Figure 5: cIAP1-cIAP2 and TRAF2 are required for receptor-induced TRAF3 degradation, which precedes NIK stabilization.
Figure 6: CD40 ligation activates cIAP1-cIAP2 through TRAF2 to induce TRAF3 ubiquitination.

Similar content being viewed by others

References

  1. Ghosh, S. & Karin, M. Missing pieces in the NF-kappaB puzzle. Cell 109 (suppl.), S81–S96 (2002).

    Article  CAS  Google Scholar 

  2. Li, Q. & Verma, I.M. NF-kappaB regulation in the immune system. Nat. Rev. Immunol. 2, 725–734 (2002).

    Article  CAS  Google Scholar 

  3. Ben-Neriah, Y. Regulatory functions of ubiquitination in the immune system. Nat. Immunol. 3, 20–26 (2002).

    Article  CAS  Google Scholar 

  4. Senftleben, U. et al. Activation by IKKalpha of a second, evolutionary conserved, NF-kappa B signaling pathway. Science 293, 1495–1499 (2001).

    Article  CAS  Google Scholar 

  5. Fong, A. & Sun, S.C. Genetic evidence for the essential role of beta-transducin repeat-containing protein in the inducible processing of NF-kappa B2/p100. J. Biol. Chem. 277, 22111–22114 (2002).

    Article  CAS  Google Scholar 

  6. Xiao, G., Harhaj, E.W. & Sun, S.C. NF-kappaB-inducing kinase regulates the processing of NF-kappaB2 p100. Mol. Cell 7, 401–409 (2001).

    Article  CAS  Google Scholar 

  7. Derudder, E. et al. RelB/p50 dimers are differentially regulated by tumor necrosis factor-alpha and lymphotoxin-beta receptor activation: critical roles for p100. J. Biol. Chem. 278, 23278–23284 (2003).

    Article  CAS  Google Scholar 

  8. Yilmaz, Z.B., Weih, D.S., Sivakumar, V. & Weih, F. RelB is required for Peyer's patch development: differential regulation of p52-RelB by lymphotoxin and TNF. EMBO J. 22, 121–130 (2003).

    Article  CAS  Google Scholar 

  9. Coope, H.J. et al. CD40 regulates the processing of NF-kappaB2 p100 to p52. EMBO J. 21, 5375–5385 (2002).

    Article  CAS  Google Scholar 

  10. Claudio, E., Brown, K., Park, S., Wang, H. & Siebenlist, U. BAFF-induced NEMO-independent processing of NF-kappa B2 in maturing B cells. Nat. Immunol. 3, 958–965 (2002).

    Article  CAS  Google Scholar 

  11. Bishop, G.A. The multifaceted roles of TRAFs in the regulation of B-cell function. Nat. Rev. Immunol. 4, 775–786 (2004).

    Article  CAS  Google Scholar 

  12. Hacker, H. & Karin, M. Regulation and function of IKK and IKK-related kinases. Sci. STKE 2006, re13 (2006).

    Article  Google Scholar 

  13. Chen, Z.J., Bhoj, V. & Seth, R.B. Ubiquitin, TAK1 and IKK: is there a connection? Cell Death Differ. 13, 687–692 (2006).

    Article  CAS  Google Scholar 

  14. 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 

  15. Shi, C.S. & Kehrl, J.H. Tumor necrosis factor (TNF)-induced germinal center kinase-related (GCKR) and stress-activated protein kinase (SAPK) activation depends upon the E2/E3 complex Ubc13-Uev1A/TNF receptor-associated factor 2 (TRAF2). J. Biol. Chem. 278, 15429–15434 (2003).

    Article  CAS  Google Scholar 

  16. Deng, L. et al. Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin-conjugating enzyme complex and a unique polyubiquitin chain. Cell 103, 351–361 (2000).

    Article  CAS  Google Scholar 

  17. Tada, K. et al. Critical roles of TRAF2 and TRAF5 in tumor necrosis factor-induced NF-kappa B activation and protection from cell death. J. Biol. Chem. 276, 36530–36534 (2001).

    Article  CAS  Google Scholar 

  18. Ishida, T. et al. Identification of TRAF6, a novel tumor necrosis factor receptor-associated factor protein that mediates signaling from an amino-terminal domain of the CD40 cytoplasmic region. J. Biol. Chem. 271, 28745–28748 (1996).

    Article  CAS  Google Scholar 

  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).

    Article  CAS  Google Scholar 

  20. Hacker, H. et al. Specificity in Toll-like receptor signalling through distinct effector functions of TRAF3 and TRAF6. Nature 439, 204–207 (2006).

    Article  Google Scholar 

  21. Rothe, M., Sarma, V., Dixit, V.M. & Goeddel, D.V. TRAF2-mediated activation of NF-kappa B by TNF receptor 2 and CD40. Science 269, 1424–1427 (1995).

    Article  CAS  Google Scholar 

  22. 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  CAS  Google Scholar 

  23. Matsuzawa, A. et al. Essential cytoplasmic translocation of a cytokine receptor-assembled signaling complex. Science 321, 663–668 (2008).

    Article  CAS  Google Scholar 

  24. Grech, A.P. et al. TRAF2 differentially regulates the canonical and noncanonical pathways of NF-kappaB activation in mature B cells. Immunity 21, 629–642 (2004).

    Article  CAS  Google Scholar 

  25. He, J.Q. et al. Rescue of TRAF3-null mice by p100 NF-kappa B deficiency. J. Exp. Med. 203, 2413–2418 (2006).

    Article  CAS  Google Scholar 

  26. Gardam, S., Sierro, F., Basten, A., Mackay, F. & Brink, R. TRAF2 and TRAF3 signal adapters act cooperatively to control the maturation and survival signals delivered to B cells by the BAFF receptor. Immunity 28, 391–401 (2008).

    Article  CAS  Google Scholar 

  27. Liao, G., Zhang, M., Harhaj, E.W. & Sun, S.C. Regulation of the NF-kappaB-inducing kinase by tumor necrosis factor receptor-associated factor 3-induced degradation. J. Biol. Chem. 279, 26243–26250 (2004).

    Article  CAS  Google Scholar 

  28. Vince, J.E. et al. IAP antagonists target cIAP1 to induce TNFalpha-dependent apoptosis. Cell 131, 682–693 (2007).

    Article  CAS  Google Scholar 

  29. Keats, J.J. et al. Promiscuous mutations activate the noncanonical NF-kappaB pathway in multiple myeloma. Cancer Cell 12, 131–144 (2007).

    Article  CAS  Google Scholar 

  30. Annunziata, C.M. et al. Frequent engagement of the classical and alternative NF-kappaB pathways by diverse genetic abnormalities in multiple myeloma. Cancer Cell 12, 115–130 (2007).

    Article  CAS  Google Scholar 

  31. Yeh, W.C. et al. Early lethality, functional NF-kappaB activation, and increased sensitivity to TNF-induced cell death in TRAF2-deficient mice. Immunity 7, 715–725 (1997).

    Article  CAS  Google Scholar 

  32. Xu, Y., Cheng, G. & Baltimore, D. Targeted disruption of TRAF3 leads to postnatal lethality and defective T-dependent immune responses. Immunity 5, 407–415 (1996).

    Article  CAS  Google Scholar 

  33. Vaux, D.L. & Silke, J. IAPs, RINGs and ubiquitylation. Nat. Rev. Mol. Cell Biol. 6, 287–297 (2005).

    Article  CAS  Google Scholar 

  34. Varfolomeev, E. et al. IAP antagonists induce autoubiquitination of c-IAPs, NF-kappaB activation, and TNFalpha-dependent apoptosis. Cell 131, 669–681 (2007).

    Article  CAS  Google Scholar 

  35. Hu, S. & Yang, X. Cellular inhibitor of apoptosis 1 and 2 are ubiquitin ligases for the apoptosis inducer Smac/DIABLO. J. Biol. Chem. 278, 10055–10060 (2003).

    Article  CAS  Google Scholar 

  36. Li, L. et al. A small molecule Smac mimic potentiates TRAIL- and TNFalpha-mediated cell death. Science 305, 1471–1474 (2004).

    Article  CAS  Google Scholar 

  37. Cohen, J.J. Glucocorticoid-induced apoptosis in the thymus. Semin. Immunol. 4, 363–369 (1992).

    CAS  PubMed  Google Scholar 

  38. Dinarello, C.A. & Mier, J.W. Lymphokines. N. Engl. J. Med. 317, 940–945 (1987).

    Article  CAS  Google Scholar 

  39. Xie, P., Stunz, L.L., Larison, K.D., Yang, B. & Bishop, G.A. Tumor necrosis factor receptor-associated factor 3 is a critical regulator of B cell homeostasis in secondary lymphoid organs. Immunity 27, 253–267 (2007).

    Article  Google Scholar 

  40. Nutt, S.L., Urbanek, P., Rolink, A. & Busslinger, M. Essential functions of Pax5 (BSAP) in pro-B cell development: difference between fetal and adult B lymphopoiesis and reduced V-to-DJ recombination at the IgH locus. Genes Dev. 11, 476–491 (1997).

    Article  CAS  Google Scholar 

  41. Igarashi, H. et al. Early lymphoid progenitors in mouse and man are highly sensitive to glucocorticoids. Int. Immunol. 17, 501–511 (2005).

    Article  CAS  Google Scholar 

  42. He, J.Q., Saha, S.K., Kang, J.R., Zarnegar, B. & Cheng, G. Specificity of TRAF3 in its negative regulation of the noncanonical NF-kappa B pathway. J. Biol. Chem. 282, 3688–3694 (2007).

    Article  CAS  Google Scholar 

  43. Rothe, M., Pan, M.G., Henzel, W.J., Ayres, T.M. & Goeddel, D.V. The TNFR2-TRAF signaling complex contains two novel proteins related to baculoviral inhibitor of apoptosis proteins. Cell 83, 1243–1252 (1995).

    Article  CAS  Google Scholar 

  44. Gallagher, E. et al. Kinase MEKK1 is required for CD40-dependent activation of the kinases Jnk and p38, germinal center formation, B cell proliferation and antibody production. Nat. Immunol. 8, 57–63 (2007).

    Article  CAS  Google Scholar 

  45. Brown, K.D., Hostager, B.S. & Bishop, G.A. Differential signaling and tumor necrosis factor receptor-associated factor (TRAF) degradation mediated by CD40 and the Epstein-Barr virus oncoprotein latent membrane protein 1 (LMP1). J. Exp. Med. 193, 943–954 (2001).

    Article  CAS  Google Scholar 

  46. Enzler, T. et al. Alternative and classical NF-kappa B signaling retain autoreactive B cells in the splenic marginal zone and result in lupus-like disease. Immunity 25, 403–415 (2006).

    Article  CAS  Google Scholar 

  47. Sivakumar, V., Hammond, K.J., Howells, N., Pfeffer, K. & Weih, F. Differential requirement for Rel/nuclear factor kappa B family members in natural killer T cell development. J. Exp. Med. 197, 1613–1621 (2003).

    Article  CAS  Google Scholar 

  48. Budanov, A.V., Sablina, A.A., Feinstein, E., Koonin, E.V. & Chumakov, P.M. Regeneration of peroxiredoxins by p53-regulated sestrins, homologs of bacterial AhpD. Science 304, 596–600 (2004).

    Article  CAS  Google Scholar 

  49. Gurova, K.V., Hill, J.E., Razorenova, O.V., Chumakov, P.M. & Gudkov, A.V. p53 pathway in renal cell carcinoma is repressed by a dominant mechanism. Cancer Res. 64, 1951–1958 (2004).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank T. Mak (University of Toronto), H. Kikutani (Osaka University) and Amgen Inc. for the Traf2+/−, Traf3+/− and Map3k14+/− mouse strains; P.M. Chumakov (Cleveland Clinic) for the pLSLPw lentiviral vector; Z. Ronai and J. Reed (Burnham Institute) for some of the expression vectors; and Santa Cruz Biotechnology for the donated antibodies. Supported by the US National Institutes of Health (M.K., D.A.A.V. and H.W.), the Kanzawa Medical Research Foundation (A.M., in part), the American Lung Association (P.-H.T., in part), a Cancer Center core grant and the American Lebanese Syrian Associated Charities (D.A.A.V. and H.W.) and the American Cancer Society (M.K.).

Author information

Authors and Affiliations

Authors

Contributions

S.V. and M.K. planned and designed all experiments and wrote the manuscript. S.V. performed most experiments. A.M. performed the ubiquitination experiments. W.Z. performed immunoprecipitation experiments. H.W. and D.A.A.V. provided Lys63-specific anti-ubiquitin. J.J.K. and P.L.B. provided the cIAP-wild-type and cIAP-deficient multiple myeloma cells. P.-H.T. made CD40-expressing HEK-293T cells.

Corresponding author

Correspondence to Michael Karin.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–8 (PDF 1288 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vallabhapurapu, S., Matsuzawa, A., Zhang, W. et al. Nonredundant and complementary functions of TRAF2 and TRAF3 in a ubiquitination cascade that activates NIK-dependent alternative NF-κB signaling. Nat Immunol 9, 1364–1370 (2008). https://doi.org/10.1038/ni.1678

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni.1678

This article is cited by

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