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Mitochondrial outer-membrane protein FUNDC1 mediates hypoxia-induced mitophagy in mammalian cells

Nature Cell Biology volume 14, pages 177–185 (2012)Cite this article

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Abstract

Accumulating evidence has shown that dysfunctional mitochondria can be selectively removed by mitophagy. Dysregulation of mitophagy is implicated in the development of neurodegenerative disease and metabolic disorders. How individual mitochondria are recognized for removal and how this process is regulated remain poorly understood. Here we report that FUNDC1, an integral mitochondrial outer-membrane protein, is a receptor for hypoxia-induced mitophagy. FUNDC1 interacted with LC3 through its typical LC3-binding motif Y(18)xxL(21), and mutation of the LC3-interaction region impaired its interaction with LC3 and the subsequent induction of mitophagy. Knockdown of endogenous FUNDC1 significantly prevented hypoxia-induced mitophagy, which could be reversed by the expression of wild-type FUNDC1, but not LC3-interaction-deficient FUNDC1 mutants. Mechanistic studies further revealed that hypoxia induced dephosphorylation of FUNDC1 and enhanced its interaction with LC3 for selective mitophagy. Our findings thus offer insights into mitochondrial quality control in mammalian cells.

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Figure 1: Mitochondrial outer-membrane protein FUNDC1 induces mitochondrial autophagy.
Figure 2: FUNDC1 as an LIR-motif-containing protein is a receptor for selective mitophagy.
Figure 3: Deletion of LIR fails to induce mitophagy, which depends on ATG5, not on Beclin 1.
Figure 4: FUNDC1 mediates mitophagy induced by hypoxia.
Figure 5: Dephosphorylation of FUNDC1 activates mitophagy induced by hypoxia.

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Acknowledgements

We wish to thank H. Zhang from the National Institute of Biological Sciences, China, and L. Yu from Tsinghua University, China, for their suggestions and critical reading of the manuscript. We thank Y. G. Chen from Tsinghua University for providing PGEX4T1-GABARAP and PGEX4T1-GABARAPL2 constructs. We wish to thank M. Bartlam from Nankai University and A. Zhou from Cleveland State University in Ohio for improvement of the English of the manuscript. The Chen laboratory was supported by the 973 project of the Ministry of Science and Technology (China) (2011CB910903 and 2010CB912204) and by grants from the National Natural Science Foundation of China (81130045, 90713006).

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  1. Lei Liu, Du Feng and Guo Chen: These authors contributed equally to this work

Authors and Affiliations

  1. The State Key Laboratory of Biomembrane and Membrane Biotechnology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China

    Lei Liu, Ming Chen, Qiaoxia Zheng, Qi Ma, Chongzhuo Zhu, Rui Wang, Baowei Li, Xiaohui Wang, Haijing Jin, Jun Wang & Quan Chen

  2. Graduate School of the Chinese Academy of Sciences, Beijing 100101, China

    Lei Liu, Ming Chen, Qiaoxia Zheng, Qi Ma, Chongzhuo Zhu, Baowei Li, Pingsheng Liu & Quan Chen

  3. College of Life Sciences, Nankai University, Tianjin 300071, China

    Du Feng, Guo Chen, Pingping Song, Wanjun Qi, Yushan Zhu & Quan Chen

  4. College of Life Sciences, Tsinghua University, Beijing 100084, China

    Lei Huang & Senfang Sui

  5. Center of Biomedical Analysis, Tsinghua University, Beijing 100084, China

    Lei Huang

  6. Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China

    Peng Xue & Fuquan Yang

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Contributions

L.L. observed that FUNDC1 could potentially induce mitophagy and carried out most of the biochemistry experiments. D.F. designed and carried out all electron microscopy analysis in S.S.’s laboratory. D.F. and G.C. identified that FUNDC1 is a substrate of Src kinase with some help from W.Q. D.F. and P.X. analysed the phosphorylation sites of FUNDC1 by mass spectrometry in F.Y.’s laboratory. D.F. and L.H. carried out the three-dimensional reconstruction and mitochondrial volume calculations. Q.Z. carried out fractionation analysis in P.L.’s laboratory. C.Z. and M.C. carried out cell-viability analysis. L.L., D.F. and Q.C. analysed and organized the data. Q.C. wrote the manuscript with input from L.L. and D.F. All authors discussed the results and commented on the manuscript.

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Correspondence to Quan Chen.

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Liu, L., Feng, D., Chen, G. et al. Mitochondrial outer-membrane protein FUNDC1 mediates hypoxia-induced mitophagy in mammalian cells. Nat Cell Biol 14, 177–185 (2012). https://doi.org/10.1038/ncb2422

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