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A ubiquitin-like system mediates protein lipidation

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Abstract

Autophagy is a dynamic membrane phenomenon for bulk protein degradation in the lysosome/vacuole1,2. Apg8/Aut7 is an essential factor for autophagy in yeast3,4,5. We previously found that the carboxy-terminal arginine of nascent Apg8 is removed by Apg4/Aut2 protease, leaving a glycine residue at the C terminus6. Apg8 is then converted to a form (Apg8-X) that is tightly bound to the membrane6. Here we report a new mode of protein lipidation. Apg8 is covalently conjugated to phosphatidylethanolamine through an amide bond between the C-terminal glycine and the amino group of phosphatidylethanolamine. This lipidation is mediated by a ubiquitination-like system. Apg8 is a ubiquitin-like protein that is activated by an E1 protein, Apg7 (refs 7, 8), and is transferred subsequently to the E2 enzymes Apg3/Aut1 (ref. 9). Apg7 activates two different ubiquitin-like proteins, Apg12 (ref. 10) and Apg8, and assigns them to specific E2 enzymes, Apg10 (ref. 11) and Apg3, respectively. These reactions are necessary for the formation of Apg8-phosphatidylethanolamine. This lipidation has an essential role in membrane dynamics during autophagy6.

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Figure 1: Characterization of molecule X.
Figure 2: Apg8 is activated by Apg7 (E1 in the Apg12 Ubl system).
Figure 3: Apg3 is a conjugating enzyme (E2) for Apg8.
Figure 4: Apg8 ubiquitination-like system is essential for autophagy.

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Acknowledgements

We would like to thank D. J. Klionsky for the anti-API antibody, A. Kihara for technical advice, and Jun-ichi Osuga for analysis of fatty acids by GC–MS. This work was supported in part by Grants-in-Aid for the Ministry of Education, Science, Culture and Sports of Japan.

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Author notes

  1. Yoshinobu Ichimura, Takayoshi Kirisako and Mariko Ohsumi: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Cell Biology, National Institute for Basic Biology, 38 Nishigonaka, Myodaijicho, 444-8585, Okazaki, Japan

    Yoshinobu Ichimura, Takayoshi Kirisako, Naotada Ishihara, Noboru Mizushima, Takeshi Noda & Yoshinori Ohsumi

  2. Department of Molecular Biomechanics, School of Life Science, The Graduate University for Advanced Studies, Okazaki, 444-8585, Japan

    Yoshinobu Ichimura, Takayoshi Kirisako, Takeshi Noda & Yoshinori Ohsumi

  3. Institute for Protein Research, Osaka University, Yamadaoka 3-2, Suita, 565-0871, Osaka, Japan

    Toshifumi Takao, Yoshinori Satomi & Yasutsugu Shimonishi

  4. PRESTO, Japan Science and Technology Corporation (JST), Kawaguchi, 332-0012, Japan

    Noboru Mizushima

  5. Department of Biosciences, High Tech Research Center, Teikyo University of Science and Technology, Yamanashi, 409-0193, Japan

    Mariko Ohsumi

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

    Isei Tanida & Eiki Kominami

Authors
  1. Yoshinobu Ichimura
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  2. Takayoshi Kirisako
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  3. Toshifumi Takao
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  4. Yoshinori Satomi
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  5. Yasutsugu Shimonishi
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  6. Naotada Ishihara
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  7. Noboru Mizushima
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  8. Isei Tanida
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  9. Eiki Kominami
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  10. Mariko Ohsumi
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  11. Takeshi Noda
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  12. Yoshinori Ohsumi
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Correspondence to Yoshinori Ohsumi.

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Ichimura, Y., Kirisako, T., Takao, T. et al. A ubiquitin-like system mediates protein lipidation. Nature 408, 488–492 (2000). https://doi.org/10.1038/35044114

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