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 ubiquitin-modifying enzyme A20 is required for termination of Toll-like receptor responses

Nature Immunology volume 5, pages 1052–1060 (2004)Cite this article

A Corrigendum to this article was published on 01 January 2005

This article has been updated

Abstract

A20 is a cytoplasmic protein required for the termination of tumor necrosis factor (TNF)–induced signals. We show here that mice doubly deficient in either A20 and TNF or A20 and TNF receptor 1 developed spontaneous inflammation, indicating that A20 is also critical for the regulation of TNF-independent signals in vivo. A20 was required for the termination of Toll-like receptor–induced activity of the transcription factor NF-κB and proinflammatory gene expression in macrophages, and this function protected mice from endotoxic shock. A20 accomplished this biochemically by directly removing ubiquitin moieties from the signaling molecule TRAF6. The critical function of this deubiquitinating enzyme in the restriction of TLR signals emphasizes the importance of the regulation of ubiquitin conjugation in innate immune cells.

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

Figure 1: A20 is essential for the prevention of TNFR1-independent proinflammatory signals.
Figure 2: A20 is essential for the prevention of TNF-independent proinflammatory signals.
Figure 3: A20 is critical for the regulation of macrophage responses in vivo and protecting against septic shock.
Figure 4: A20 regulates macrophage responses to TLR ligands.
Figure 5: A20 is essential for the termination of TLR-induced IKK activity.
Figure 6: A20 is not required for endotoxin tolerance.
Figure 7: A20 is a deubiquitinating enzyme that modifies TRAF6 ubiquitination.

Similar content being viewed by others

Change history

  • 12 September 2004

    added footnote to XML; appended incorrect AOP PDF with note; added notes in xml for all Fig 7 instances; corrected online date will be added to issue version

Notes

  1. *Note: In the version of this article originally published online, the last sentence of the legend to Figure 7 was incorrect and should be deleted. This error has been corrected for the HTML and print versions of this article.

References

  1. Takeda, K., Kaisho, T. & Akira, S. Toll-like receptors. Annu. Rev. Immunol. 21, 335–376 (2003).

    Article  CAS  Google Scholar 

  2. Janeway, C.A., Jr. & Medzhitov, R. Innate immune recognition. Annu. Rev. Immunol. 20, 197–216 (2002).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  4. Barton, G.M. & Medzhitov, R. Toll-like receptor signaling pathways. Science 300, 1524–1525 (2003).

    Article  CAS  Google Scholar 

  5. Akira, S. Toll-like receptor signaling. J. Biol. Chem. 278, 38105–38108 (2003).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  9. Zhang, G. & Ghosh, S. Negative regulation of Toll-like receptor-mediated signaling by Tollip. J. Biol. Chem. 277, 7059–7065 (2002).

    Article  CAS  Google Scholar 

  10. 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 

  11. Brint, E. et al. ST2 is an inhibitor of interleukin 1 receptor and Toll-like receptor 4 signaling and maintains endotoxin tolerance. Nat. Immunol. 5, 373–379 (2004).

    Article  CAS  Google Scholar 

  12. Chuang, T.-H. & Ulevitch, R. Triad3A, an E3 ubiquitin-protein ligase regulating Toll-like receptors. Nat. Immunol. 5, 495–502 (2004).

    Article  CAS  Google Scholar 

  13. Opipari, A.W., Jr., Boguski, M.S. & Dixit, V.M. The A20 cDNA induced by tumor necrosis factor α encodes a novel type of zinc finger protein. J. Biol. Chem. 265, 14705–14708 (1990).

    CAS  PubMed  Google Scholar 

  14. Krikos, A., Laherty, C.D. & Dixit, V.M. Transcriptional activation of the tumor necrosis factor α-inducible zinc finger protein, A20, is mediated by κB elements. J. Biol. Chem. 267, 17971–17976 (1992).

    CAS  PubMed  Google Scholar 

  15. Lee, E.G. et al. Failure to regulate TNF-induced NF-κB and cell death responses in A20-deficient mice. Science 289, 2350–2354 (2000).

    Article  CAS  Google Scholar 

  16. Heyninck, K. & Beyaert, R. The cytokine-inducible zinc finger protein A20 inhibits IL-1-induced NF-κB activation at the level of TRAF6. FEBS Lett. 442, 147–150 (1999).

    Article  CAS  Google Scholar 

  17. Cooper, J.T. et al. A20 blocks endothelial cell activation through a NF-κB-dependent mechanism. J. Biol. Chem. 271, 18068–18073 (1996).

    Article  CAS  Google Scholar 

  18. Nau, G.J. et al. Human macrophage activation programs induced by bacterial pathogens. Proc. Natl. Acad. Sci. USA 99, 1503–1508 (2002).

    Article  CAS  Google Scholar 

  19. Nomura, F. et al. Cutting edge: endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface Toll-like receptor 4 expression. J. Immunol. 164, 3476–3479 (2000).

    Article  CAS  Google Scholar 

  20. Makarova, K.S., Aravind, L. & Koonin, E.V. A novel superfamily of predicted cysteine proteases from eukaryotes, viruses and Chlamydia pneumoniae. Trends Biochem. Sci. 25, 50–52 (2000).

    Article  CAS  Google Scholar 

  21. Evans, P.C. et al. A novel type of deubiquitinating enzyme. J. Biol. Chem. 278, 23180–23186 (2003).

    Article  CAS  Google Scholar 

  22. Borodovsky, A. et al. Chemistry-based functional proteomics reveals novel members of the deubiquitinating enzyme family. Chem. Biol. 9, 1149–1159 (2002).

    Article  CAS  Google Scholar 

  23. Balakirev, M.Y., Tcherniuk, S.O., Jaquinod, M. & Chroboczek, J. Otubains: a new family of cysteine proteases in the ubiquitin pathway. EMBO Rep. 4, 517–522 (2003).

    Article  CAS  Google Scholar 

  24. Weissman, A.M. Themes and variations on ubiquitylation. Nat. Rev. Mol. Cell. Biol. 2, 169–178 (2001).

    Article  CAS  Google Scholar 

  25. 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 

  26. Wang, C. et al. TAK1 is a ubiquitin-dependent kinase of MKK and IKK. Nature 412, 346–351 (2001).

    Article  CAS  Google Scholar 

  27. Deng, L. et al. Activation of the IκB 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 

  28. Baud, V. et al. Signaling by proinflammatory cytokines: oligomerization of TRAF2 and TRAF6 is sufficient for JNK and IKK activation and target gene induction via an amino-terminal effector domain. Genes Dev. 13, 1297–1308 (1999).

    Article  CAS  Google Scholar 

  29. Ohashi, K., Burkart, V., Flohe, S. & Kolb, H. Cutting edge: heat shock protein 60 is a putative endogenous ligand of the Toll-like receptor-4 complex. J. Immunol. 164, 558–561 (2000).

    Article  CAS  Google Scholar 

  30. Okamura, Y. et al. The extra domain A of fibronectin activates Toll-like receptor 4. J. Biol. Chem. 276, 10229–10233 (2001).

    Article  CAS  Google Scholar 

  31. Evans, P.C. et al. Zinc-finger protein A20, a regulator of inflammation and cell survival, has de-ubiquitinating activity. Biochem. J. 378, 727–734 (2004).

    Article  CAS  Google Scholar 

  32. Beyaert, R, Heyninck, K & VanHuffel, S. A20 and A20-binding proteins as cellular inhibitors of nuclear factor-κB-dependent gene expression and apoptosis. Biochem. Pharmacol. 60, 1143–1151 (2000).

    Article  CAS  Google Scholar 

  33. Song, H.Y., Rothe, M. & Goeddel, D.V. The tumor necrosis factor-inducible zinc finger protein A20 interacts with TRAF1/TRAF2 and inhibits NF-κB activation. Proc. Natl. Acad. Sci. USA 93, 6721–6725 (1996).

    Article  CAS  Google Scholar 

  34. Brummelkamp, T.R., Nijman, S.M., Dirac, A.M. & Bernards, R. Loss of the cylindromatosis tumour suppressor inhibits apoptosis by activating NF-κB. Nature 424, 797–801 (2003).

    Article  CAS  Google Scholar 

  35. Kovalenko, A. et al. The tumour suppressor CYLD negatively regulates NF-κB signalling by deubiquitination. Nature 424, 801–805 (2003).

    Article  CAS  Google Scholar 

  36. Trompouki, E. et al. CYLD is a deubiquitinating enzyme that negatively regulates NF-κB activation by TNFR family members. Nature 424, 793–796 (2003).

    Article  CAS  Google Scholar 

  37. Bignell, G.R. et al. Identification of the familial cylindromatosis tumour-suppressor gene. Nat. Genet. 25, 160–165 (2000).

    Article  CAS  Google Scholar 

  38. Wertz, I.E. et al. De-ubiquitination and ubiquitin ligase domains of A20 downregulate NF-κB signalling. Nature 430, 694–699 (2004).

    Article  CAS  Google Scholar 

  39. Burkett, P. et al. IL-15Rα expression on CD8+ T cells is dispensable for T cell memory. Proc. Natl. Acad. Sci. USA 100, 4724–4729 (2003).

    Article  CAS  Google Scholar 

  40. Chutkow, W.A. et al. Episodic coronary artery vasospasm and hypertension develop in the absence of Sur2 K(ATP) channels. J. Clin. Inv. 110, 203W. A. 208 (2002).

  41. Hofmann, R.M. & Pickart, C.M. In vitro assembly and recognition of Lys-63 polyubiquitin chains. J. Biol. Chem. 276, 27936–27943 (2001).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank I. Wertz, V. Dixit, and B. Malynn for reagents, discussions and critical reading of the manuscript. Supported by the National Institutes of Health (RO1AI53224 and R01DK52751 to A.M.), Digestive Disease Research Center (DK42086), Burroughs Welcome (E.M.), Crohn's and Colitis Foundation of America (D.B.) and Medical Scientist Training Program (GM07281; E.T., R.-C.A. and M.W.).

Author information

Author notes

  1. David L Boone, Emre E Turer and Eric G Lee: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medicine, University of California at San Francisco, San Francisco, 94143-0451, California, USA

    David L Boone, Emre E Turer, Eric G Lee, Regina-Celeste Ahmad, Paula Hurley, Marcia Chien, Sophia Chai, Osamu Hitotsumatsu & Averil Ma

  2. Department of Medicine, University of Chicago, Chicago, 60637, Illinois, USA

    Matthew T Wheeler & Elizabeth McNally

  3. Department of Biochemistry and Molecular Biology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, 21205, Maryland, USA

    Colleen Tsui & Cecile Pickart

Authors
  1. David L Boone
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Emre E Turer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eric G Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Regina-Celeste Ahmad
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Matthew T Wheeler
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Colleen Tsui
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Paula Hurley
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Marcia Chien
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Sophia Chai
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Osamu Hitotsumatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Elizabeth McNally
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Cecile Pickart
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Averil Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Averil Ma.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Boone, D., Turer, E., Lee, E. et al. The ubiquitin-modifying enzyme A20 is required for termination of Toll-like receptor responses. Nat Immunol 5, 1052–1060 (2004). https://doi.org/10.1038/ni1110

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ni1110

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