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Smad6 negatively regulates interleukin 1-receptor–Toll-like receptor signaling through direct interaction with the adaptor Pellino-1

Nature Immunology volume 7, pages 1057–1065 (2006)Cite this article

Abstract

Transforming growth factor-β1 (TGF-β1) is a potent cytokine with pleiotropic effects, including anti-inflammatory activity. Here we show that the signaling protein Smad6 bound to Pellino-1, an adaptor protein of mammalian interleukin 1 receptor (IL-1R)–associated kinase 1 (IRAK1), and thereby promoted TGF-β-mediated anti-inflammatory effects. Smad6–Pellino-1 interaction abrogated signaling mediated by a complex of IRAK1, Pellino-1 and adaptor protein TRAF6 that formed after stimulation by IL-1β treatment. Blockade of IRAK1–Pellino-1–TRAF6 signaling prevented degradation of the inhibitor IκBα and subsequent nuclear translocation of transcription factor NF-κB and thus expression of proinflammatory genes. 'Knockdown' of endogenous Smad6 expression by RNA interference reduced anti-inflammatory activity mediated by TGF-β1 or the TGF-β family member BMP-4. Thus Smad6 is a critical mediator of the TGF-β–BMP pathway that mediates anti-inflammatory activity and negatively regulates IL-1R–Toll-like receptor signals.

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Figure 1: Smad6 binds to Pellino-1.
Figure 2: Smad6 blocks IRAK1-mediated signaling complex formation induced by IL-1β treatment.
Figure 3: Smad6 inhibits NF-κB-mediated reporter gene induction by IL-1β or LPS.
Figure 4: Smad6 inhibits IκBα degradation and nuclear translocation of NF-κB.
Figure 5: Expression of Smad6 reduces the expression of proinflammatory genes induced by IL-1β treatment.
Figure 6: Inhibition of endogenous Smad6 reduces the anti-inflammatory effects of TGF-β1 or BMP-4.
Figure 7: Smad6 induction by TGF-β1 is required for the inhibition of proinflammatory gene expression.

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Acknowledgements

We thank G. Merlino and A. Hobbie for critical reading of the manuscript. Supported by Korea Research Foundation Grant funded by the Korean Government (KRF-2003-015-C00528 to S.H.P.; laboratory equipment, R08-2003-000-10077-0 to S.H.P.), the intramural research fund of the National Cancer Center of Korea (C.-H.L. and I.-H.K) and the Intramural Research Program of the National Cancer Institute.

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Author notes

  1. Kyung-Chul Choi and Youn Sook Lee: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, Inha University College of Medicine and Inha Research Institute for Medical Sciences, Inha University College of Medicine by BK-21 Project, Incheon, 400-712, Republic of Korea

    Kyung-Chul Choi, Youn Sook Lee, Hyo Kyoung Choi, Eun-Kyung Lee & Seok Hee Park

  2. Center for Advanced Medical Education, Inha University College of Medicine by BK-21 Project, Incheon, 400-712, Republic of Korea

    Youn Sook Lee, Eun-Kyung Lee & Seok Hee Park

  3. Laboratory of Cell Regulation and Carcinogenesis, National Cancer Institute, National Institute of Health, Bethesda, 20892, Maryland, USA

    Seunghwan Lim, Suntaek Hong & Seong-Jin Kim

  4. Research Institute, National Cancer Center, Goyang, 411-769, Gyeonggi, Republic of Korea

    Chang-Hun Lee & In-Hoo Kim

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Contributions

K.-C.C., Y.S.L., S.L., H.K.C., C.-H.L., E.-K.L. and S.H. did the experimental work and analyzed data; I.-H.K. participated in the study design; S.H.P. and S.-J.K. designed and conceptualized the research, supervised the experimental work, analyzed data and wrote the manuscript.

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Correspondence to Seong-Jin Kim or Seok Hee Park.

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Supplementary information

Supplementary Fig. 1

Smad6 inhibits the expression of the NF-κB-promoter reporter induced by LPS in RAW264.7 cells. (PDF 687 kb)

Supplementary Fig. 2

Schematic representation of Smad6 'knockdown' plasmid (pU6-siSmad6) by siRNA. (PDF 757 kb)

Supplementary Fig. 3

Inhibition of endogenous Pellino-1 by siRNA abrogates NF-κB activation induced by IL-1β treatment. (PDF 711 kb)

Supplementary Fig. 4

Stable expression of Peli1 siRNA decreases NF-κB activation upon treatment of IL-1β. (PDF 186 kb)

Supplementary Fig. 5

LPS induced IRAK1-mediated signaling complexes are disrupted by pre-treatment with TGF-β1. (PDF 201 kb)

Supplementary Fig. 6

IL-1β-induced endogenous TRAF6-mediated signaling complexes are disrupted by pre-treatment with TGF-β1. (PDF 199 kb)

Supplementary Fig. 7

Models for the negative regulation of IL-1R–TLR signaling by Smad6. (PDF 201 kb)

Supplementary Table 1

Primer sequences. (PDF 33 kb)

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Choi, KC., Lee, Y., Lim, S. et al. Smad6 negatively regulates interleukin 1-receptor–Toll-like receptor signaling through direct interaction with the adaptor Pellino-1. Nat Immunol 7, 1057–1065 (2006). https://doi.org/10.1038/ni1383

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