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ROS-dependent activation of the TRAF6-ASK1-p38 pathway is selectively required for TLR4-mediated innate immunity

Nature Immunology volume 6, pages 587–592 (2005)Cite this article

Abstract

Apoptosis signal–regulating kinase 1 (ASK1) is an evolutionarily conserved mitogen-activated protein 3-kinase that activates both Jnk and p38 mitogen-activated protein kinases. Here we used ASK1-deficient mice to show that ASK1 was selectively required for lipopolysaccharide-induced activation of p38 but not of Jnk or the transcription factor NF-κB. ASK1 was required for the induction of proinflammatory cytokines dependent on Toll-like receptor 4 (TLR4) but not TLR2 or other TLRs. Consistent with this, ASK1-deficient mice were resistant to lipopolysaccharide-induced septic shock. Lipopolysaccharide induced the production of intracellular reactive oxygen species, which was required for the formation of a complex of the adaptor molecule TRAF6 and ASK1 and subsequent activation of the ASK1-p38 pathway. Our data demonstrate that the reactive oxygen species–dependent TRAF6-ASK1-p38 axis is crucial for TLR4-mediated mammalian innate immunity.

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Figure 1: ASK1 is selectively required for LPS-induced p38 but not Jnk or NF-κB activation.
Figure 2: ASK1 is required for LPS-induced production of inflammatory factors.
Figure 3: ASK1 is selectively required for signaling dependent on TLR4 but not TLR2, TLR3 or TLR9.
Figure 4: LPS-induced activation of ASK1-p38 signaling and production of inflammatory cytokine are mediated by ROS production.
Figure 5: ROS-dependent formation of the TRAF6-ASK1 complex is involved in MyD88-dependent LPS signaling.
Figure 6: ASK1-deficient mice are resistant to LPS-induced septic shock.

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Acknowledgements

We thank M. Karin and T. Taga for comments, and all the members of Laboratory of Cell Signaling. Supported by Grants-in-Aid for scientific research from the Ministry of Education, Culture, Sports, Science and Technology of Japan and Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation.

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Author notes

  1. Atsushi Matsuzawa and Kaoru Saegusa: These authors contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Cell Signaling, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Atsushi Matsuzawa, Kaoru Saegusa, Takuya Noguchi, Chiharu Sadamitsu, Hideki Nishitoh, Kohsuke Takeda & Hidenori Ichijo

  2. Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Atsushi Matsuzawa, Kaoru Saegusa, Takuya Noguchi, Chiharu Sadamitsu, Hideki Nishitoh, Shigenori Nagai, Shigeo Koyasu, Kunihiro Matsumoto, Kohsuke Takeda & Hidenori Ichijo

  3. Strategic Approach to Drug Discovery and Development in Pharmaceutical Sciences, Center of Excellence Program, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan

    Atsushi Matsuzawa, Kaoru Saegusa, Takuya Noguchi, Chiharu Sadamitsu, Hideki Nishitoh, Kohsuke Takeda & Hidenori Ichijo

  4. Laboratory of Cell Signaling, Graduate School of Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8549, Japan

    Takuya Noguchi & Hideki Nishitoh

  5. Department of Microbiology and Immunology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan

    Shigenori Nagai & Shigeo Koyasu

  6. Department of Molecular Biology, Group of Signal Transduction, Graduate School of Science, Nagoya University, Furou-chou, Chikusa-ku, Nagoya, 464-8602, Aichi, Japan

    Kunihiro Matsumoto

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Correspondence to Hidenori Ichijo.

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Supplementary information

Supplementary Fig. 1

LPS-induced expression of inducible NO synthase (iNOS) is reduced in ASK1-deficient DCs. (PDF 70 kb)

Supplementary Fig. 2

Normal expression of TLR4–MD-2 in ASK1-deficient cells and inhibitory effects of p38 inhibitor on LPS-induced cytokine productions. (PDF 110 kb)

Supplementary Fig. 3

PGN-induced IL-6 production in ASK1-deficient splenocytes and inability of poly(I:C) and CpG on ASK1 activation. (PDF 312 kb)

Supplementary Fig. 4

ASK1 is not essential for TLR2-dependent pathway. (PDF 199 kb)

Supplementary Fig. 5

ROS-dependent endogenous interaction between TRAF6 and ASK1 induced by LPS. (PDF 199 kb)

Supplementary Fig. 6

Specificity of TRAF6 antibody (PDF 142 kb)

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Matsuzawa, A., Saegusa, K., Noguchi, T. et al. ROS-dependent activation of the TRAF6-ASK1-p38 pathway is selectively required for TLR4-mediated innate immunity. Nat Immunol 6, 587–592 (2005). https://doi.org/10.1038/ni1200

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