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FLIP-mediated autophagy regulation in cell death control

Nature Cell Biology volume 11, pages 1355–1362 (2009)Cite this article

Abstract

Autophagy is an active homeostatic degradation process for the removal or turnover of cytoplasmic components wherein the LC3 ubiquitin-like protein undergoes an Atg7 E1-like enzyme/Atg3 E2-like enzyme-mediated conjugation process to induce autophagosome biogenesis1,2,3,4. Besides its cytoprotecive role, autophagy acts on cell death when it is abnormally upregulated. Thus, the autophagy pathway requires tight regulation to ensure that this degradative process is well balanced. Two death effector domains (DED1/2) containing cellular FLICE-like inhibitor protein (cFLIP) and viral FLIP (vFLIP) of Kaposi's sarcoma-associated herpesvirus (KSHV), Herpesvirus saimiri (HVS), and Molluscum contagiosum virus (MCV) protect cells from apoptosis mediated by death receptors5,6. Here, we report that cellular and viral FLIPs suppress autophagy by preventing Atg3 from binding and processing LC3. Consequently, FLIP expression effectively represses cell death with autophagy, as induced by rapamycin, an mTor inhibitor and an effective anti-tumour drug against KSHV-induced Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL)7,8. Remarkably, either a DED1 α2-helix ten amino-acid (α2) peptide or a DED2 α4-helix twelve amino-acid (α4) peptide of FLIP is individually sufficient for binding FLIP itself and Atg3, with the peptide interactions effectively suppressing Atg3–FLIP interaction without affecting Atg3-LC3 interaction, resulting in robust cell death with autophagy. Our study thus identifies a checkpoint of the autophagy pathway where cellular and viral FLIPs limit the Atg3-mediated step of LC3 conjugation to regulate autophagosome biogenesis. Furthermore, the FLIP-derived short peptides induce growth suppression and cell death with autophagy, representing biologically active molecules for potential anti-cancer therapies.

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Figure 1: FLIP suppresses autophagy.
Figure 2: FLIP interacts with Atg3.
Figure 3: vFLIP blocks growth suppression and cell death with autophagy induced by rapamycin.
Figure 4: FLIP α2 and α4 peptides induce cell death with autophagy.
Figure 5: The vFLIP peptide binds FLIP at higher affinity and peptide binding blocks Atg3–FLIP interaction without affecting Atg3–LC3 interaction.

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Acknowledgements

This work was partly supported by US. Public Health Service grants CA82057, CA91819, CA31363, CA115284, AI073099, RR00168, Hastings Foundation, Korean GRL Program K20815000001 (J.J.), AI083841, CA140964, the Lymphoma and Leukemia Society of USA, the Wright Foundation, and the Baxter Foundation (C.L.). We thank Don Ganem, Noboru Mizushima, Preet Chaudhary and Jeff Cohen for reagents, Ernesto Barron for EM analysis, and Steven Lee for manuscript preparation. Finally, we thank all of J.J.'s lab members for their discussions.

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Authors and Affiliations

  1. Department of Molecular Microbiology and Immunology, University of Southern California, Keck School of Medicine, Harlyne J. Norris Cancer Research Tower, 1450 Biggy Street, Los Angeles, 90033, California, USA

    Jong-Soo Lee, June-Yong Lee, Sun-Hwa Lee, Joseph H. Jeong, Hye-Ra Lee, Heesoon Chang, Chengyu Liang & Jae U. Jung

  2. Department of Microbiology and Molecular Genetics and Tumour Virology Division, New England Primate Research Center, Harvard Medical School, 1 Pine Hill Drive, Southborough, 01772, Massachusetts, USA

    Jong-Soo Lee, Qinglin Li, Hye-Ra Lee, Heesoon Chang, Chengyu Liang & Jae U. Jung

  3. Department of Pediatrics, The University of Texas Health Science Center at San Antonio, 78229, USA

    Fu-Chun Zhou & Shou-Jiang Gao

  4. Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute and the Department of Pathology, Harvard Medical School, Boston, 02115, MA

    Qinglin Li

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Contributions

J.S.L. performed all aspects of this study; Q.L. performed the initial study; J.Y.L, S.H.L, H.J, and H.R.L assisted with the experimental design and in collecting the data; F.C.Z and S.J.G. provided KSHV ΔvFLIP; C.L. assisted with the experimental design and interpretation; J.S.L. and J.J. organized this study and wrote the paper. All authors discussed the results and commented on the manuscript.

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Correspondence to Jae U. Jung.

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Lee, JS., Li, Q., Lee, JY. et al. FLIP-mediated autophagy regulation in cell death control. Nat Cell Biol 11, 1355–1362 (2009). https://doi.org/10.1038/ncb1980

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