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The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1

Nature Cell Biology volume 12, pages 213–223 (2010)Cite this article

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Abstract

Impaired selective turnover of p62 by autophagy causes severe liver injury accompanied by the formation of p62-positive inclusions and upregulation of detoxifying enzymes. These phenotypes correspond closely to the pathological conditions seen in human liver diseases, including alcoholic hepatitis and hepatocellular carcinoma. However, the molecular mechanisms and pathophysiological processes in these events are still unknown. Here we report the identification of a novel regulatory mechanism by p62 of the transcription factor Nrf2, whose target genes include antioxidant proteins and detoxification enzymes. p62 interacts with the Nrf2-binding site on Keap1, a component of Cullin-3-type ubiquitin ligase for Nrf2. Thus, an overproduction of p62 or a deficiency in autophagy competes with the interaction between Nrf2 and Keap1, resulting in stabilization of Nrf2 and transcriptional activation of Nrf2 target genes. Our findings indicate that the pathological process associated with p62 accumulation results in hyperactivation of Nrf2 and delineates unexpected roles of selective autophagy in controlling the transcription of cellular defence enzyme genes.

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Figure 1: Interaction between Keap1 and p62.
Figure 2: Crystal structure of Keap1-DC in complex with p62-KIR.
Figure 3: Competitive inhibition of the Nrf2–Keap1 pathway by p62.
Figure 4: Formation of p62-positive and Keap1-positive inclusions in autophagy-deficient hepatocytes.
Figure 5: Amelioration of liver dysfunction in autophagy-deficient mice by the additional loss of Nrf2.
Figure 6: Exacerbation of liver dysfunction in autophagy-deficient mice by the additional loss of Keap1.

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Acknowledgements

We thank T. Kouno and K. Endo (Tokyo Metropolitan Institute of Medical Science) for technical assistance, and the beamline staff at NW12 of PF-AR (Tsukuba, Japan) for technical help in data collection. We also thank A. Yamada, K. Kanno and A. Yabashi (Fukushima Medical University School of Medicine) for their help in histological studies; J. Yanagisawa (Tsukuba University) and Y. Saeki (Tokyo Metropolitan Institute of Medical Science) for mass spectrometric analyses; Y. Kawatani (Tohoku University) for technical assistance in microarray analyses; and R. Kopito and B. E. Riley (Stanford University) and T. Mizushima and T. Kumanomidou (Nagoya University) for helpful discussion. p62-knockout mice were provided by T. Ishii (Tsukuba University). The Biomedical Research Core of Tohoku University Graduate School of Medicine provided DRI-CHEM 7000V (Fuji Film Corp.). This work was supported by grants from the Japan Science and Technology Agency (M.K.), the Ministry of Education, Science and Culture of Japan (M.K., K.T. and M.Y.) and the Targeted Proteins Research Program (H.K., K.T. and M.Y.).

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Authors and Affiliations

  1. Laboratory of Frontier Science, Tokyo Metropolitan Institute of Medical Science, Bunkyo-ku, 113-8613, Tokyo, Japan

    Masaaki Komatsu, Yoshinobu Ichimura, Yu-Shin Sou, Izumi Ueno, Ayako Sakamoto, Yasumasa Nishito & Keiji Tanaka

  2. PRESTO, Japan Science and Technology Corporation, 332-0012, Kawaguchi, Japan

    Masaaki Komatsu

  3. Department of Medical Biochemistry and ERATO-JST, Tohoku University Graduate School of Medicine, Aoba-ku, 980-8575, Sendai, Japan

    Hirofumi Kurokawa, Keiko Taguchi, Kit I. Tong, Hozumi Motohashi & Masayuki Yamamoto

  4. Department of Anatomy and Histology, Fukushima Medical University School of Medicine, Hikarigaoka, 960-1295, Fukushima, Japan

    Satoshi Waguri

  5. Department of Medical Life Systems, Doshisha University, Kyoutanabe, 610-0394, Kyoto, Japan

    Akira Kobayashi & Mihee Kim

  6. Department of Biochemistry, Juntendo University School of Medicine, Bunkyo-ku, 113-8421, Tokyo, Japan

    Yoshinobu Ichimura, Yu-Shin Sou, Takashi Ueno & Eiki Kominami

  7. National Institutes of Advanced Industrial Science and Technology, Biological Information Research Center (JBIRC), Kohtoh-ku, 135-0064, Tokyo, Japan

    Shun-ichiro Iemura & Tohru Natsume

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Contributions

M.K., K.T., I.U. and A.S. performed most of the experiments that characterized the knockout mice. I.U., Y.-S.S., Y.I. and A.K. performed the biochemical and cell biological experiments. Y.N. carried out microarray analyses. S.-i.I and T.N. performed mass spectrometric analyses. S.W. performed the histological and microscopic analyses. Structural and kinetics analyses were completed by H.K. and K.I.T. M.K., K.T. and M.Y. conceived the experiments. M.K., H.K., S.W., H.M., K.T. and M.Y. wrote the paper. E.K. and T.U. provided intellectual support. All authors discussed the results and commented on the manuscript.

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Correspondence to Masaaki Komatsu, Keiji Tanaka or Masayuki Yamamoto.

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Komatsu, M., Kurokawa, H., Waguri, S. et al. The selective autophagy substrate p62 activates the stress responsive transcription factor Nrf2 through inactivation of Keap1. Nat Cell Biol 12, 213–223 (2010). https://doi.org/10.1038/ncb2021

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