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Different modes of ubiquitination of the adaptor TRAF3 selectively activate the expression of type I interferons and proinflammatory cytokines

Nature Immunology volume 11, pages 70–75 (2010)Cite this article

Abstract

Balanced production of type I interferons and proinflammatory cytokines after engagement of Toll-like receptors (TLRs), which signal through adaptors containing a Toll–interleukin 1 receptor (TIR) domain, such as MyD88 and TRIF, has been proposed to control the pathogenesis of autoimmune disease and tumor responses to inflammation. Here we show that TRAF3, a ubiquitin ligase that interacts with both MyD88 and TRIF, regulated the production of interferon and proinflammatory cytokines in different ways. Degradative ubiquitination of TRAF3 during MyD88-dependent TLR signaling was essential for the activation of mitogen-activated protein kinases (MAPKs) and production of inflammatory cytokines. In contrast, TRIF-dependent signaling triggered noncanonical TRAF3 self-ubiquitination that activated the interferon response. Inhibition of degradative ubiquitination of TRAF3 prevented the expression of all proinflammatory cytokines without affecting the interferon response.

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Figure 1: Role of cIAP1/2 in TLR-mediated MAPK signaling.
Figure 2: TLR4 engagement induces an MyD88-associated signaling complex that undergoes cIAP1/2- and TRAF6-dependent cytosolic translocation after TRAF3 degradation.
Figure 3: TRAF6 is required for LPS-induced activation of TAK1 and ubiquitination of cIAP2 and TRAF3.
Figure 4: LPS-induced K63-linked TRAF3 self-ubiquitination depends on TLR endocytosis.
Figure 5: K63- and K48-linked ubiquitination have different and distinct roles in TRAF3 function.
Figure 6: Differences in the regulation of TLR4-induced inflammatory cytokines and interferon-related genes.

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Acknowledgements

We thank H. Ichijo (University of Tokyo) for providing A.M. with space and support for some of this work described above; S. Akira (Osaka University) for Myd88−/− mice; B. Beutler (Scripps Research Institute) for TrifLps2/Lps2 mice; R. Fonseca (Mayo Clinic) for multiple myeloma cells; X. Wang (University of Texas Southwestern Medical Center) for SM; H. Wang (St. Jude Children's Research Hospital) for generating monoclonal antibody HWA4C4, specific for K63-linked ubiquitin; I. Verma (Salk Institute) for pLV-CMV-delta 8.2; and Millipore for the antibody to K48-linked polyubiquitin. Supported by the National Institutes of Health (AI043477 to M.K. and AI52199 to D.A.A.V.), the American Cancer Society (M.K.), the American Lung Association of California (P.-H.T.), the Global Center of Excellence program (A.M.), the Toyobo Biotechnology Foundation (T.M.), the National Cancer Institute (CA21765 to D.A.A.V.) and the American Lebanese Syrian Associated Charities (D.A.A.V.).

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Author notes

  1. Ping-Hui Tseng and Atsushi Matsuzawa: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology and Department of Pathology, Laboratory of Gene Regulation and Signal Transduction, School of Medicine, University of California San Diego, La Jolla, California, USA

    Ping-Hui Tseng, Atsushi Matsuzawa, Weizhou Zhang, Takashi Mino & Michael Karin

  2. Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan

    Ping-Hui Tseng

  3. Laboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, Japan

    Atsushi Matsuzawa

  4. Department of Immunology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    Dario A A Vignali

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  1. Ping-Hui Tseng
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Contributions

P.-H.T. and M.K. planned and designed all experiments and wrote the manuscript; P.-H.T. and A.M. did most experiments; W.Z. and T.M. helped with cell cultures, TRAF3 mutants and immunoprecipitation experiments; and D.A.A.V. provided the HWA4C4 K63-specific antibody to ubiquitin.

Corresponding author

Correspondence to Michael Karin.

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Tseng, PH., Matsuzawa, A., Zhang, W. et al. Different modes of ubiquitination of the adaptor TRAF3 selectively activate the expression of type I interferons and proinflammatory cytokines. Nat Immunol 11, 70–75 (2010). https://doi.org/10.1038/ni.1819

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