FLIP-mediated autophagy regulation in cell death control (original) (raw)

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.

This is a preview of subscription content, access via your institution

Access options

Subscribe to this journal

Receive 12 print issues and online access

$259.00 per year

only $21.58 per issue

Buy this article

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Additional access options:

Similar content being viewed by others

References

  1. Levine, B. & Kroemer, G. Autophagy in the pathogenesis of disease. Cell 132, 27–42 (2008).
    Article CAS Google Scholar
  2. Mizushima, N., Levine, B., Cuervo, A. M. & Klionsky, D. J. Autophagy fights disease through cellular self-digestion. Nature 451, 1069–1075 (2008).
    Article CAS Google Scholar
  3. Geng, J & Klionsky, D, J. The Atg8 and Atg12 ubiquitin-like conjugation systems in macroautophagy. 'Protein Modifications: Beyond the Usual Suspects' Review Series. EMBO Rep. 9, 859–864 (2008).
    Article CAS Google Scholar
  4. Mizushima, N. Two ubiquitin-like conjugation systems essential for autophagy. Sem. Cell Dev. Biol. 15, 231–236 (2004).
    Article Google Scholar
  5. Thome, M. & Tschopp, J. Regulation of lymphocyte proliferation and death by FLIP. Nature Rev. Immunol. 1, 50–58 (2001).
    Article CAS Google Scholar
  6. Li, F. Y., Jeffrey, P. D., Yu, J.W & Shi, Y. Crystal structure of a viral FLIP. J. Biol. Chem. 281, 2960–2968 (2006).
    Article CAS Google Scholar
  7. Izzedine, H., Brocheriou, I. & Frances, C. Post-transplantation proteinuria and sirolimus. N. Engl. J. Med. 353, 2088–2089 (2005).
    Article CAS Google Scholar
  8. Sin, S. H. et al. Rapamycin is efficacious against primary effusion lymphoma (PEL) cell lines in vivo by inhibiting autocrine signaling. Blood 109, 2165–2173 (2007).
    Article CAS Google Scholar
  9. Levine, B. & Klionsky, D. J. Development by self-digestion: molecular mechanisms and biological functions of autophagy. Dev. Cell 6, 463–477 (2004).
    Article CAS Google Scholar
  10. Nakatogawa H, Ichimura Y, Ohsumi Y . Atg8, a ubiquitin-like protein required for autophagosome formation, mediates membrane tethering and hemifusion. Cell 13, 165–178 (2007).
    Article Google Scholar
  11. Ku, B. et al. Structural and biochemical bases for the inhibition of autophagy and apoptosis by viral BCL-2 of murine gamma-herpesvirus 68. PLoS Pathog. 4, e25 (2008).
    Article Google Scholar
  12. Liang, C. et al. Autophagic and tumour suppressor activity of a novel Beclin1-binding protein UVRAG. Nature Cell. Biol. 8, 688–699 (2006).
    Article CAS Google Scholar
  13. Pattingre, S. et al. Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 122, 927–939 (2005).
    Article CAS Google Scholar
  14. Mizushima, N., Yamamoto, A., Matsui, M., Yoshimori, T. & Ohsumi, Y. In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker. Mol. Biol. Cell 15, 1101–1111 (2004).
    Article CAS Google Scholar
  15. Nakamura, H. et al. Global changes in Kaposi's sarcoma-associated virus gene expression patterns following expression of a tetracycline-inducible Rta transactivator. J. Virol. 77, 4205–4220 (2003).
    Article CAS Google Scholar
  16. Guasparri, I., Wu, H. & Cesarman, E. The KSHV oncoprotein vFLIP contains a TRAF-interacting motif and requires TRAF2 and TRAF3 for signalling. EMBO Rep. 7, 114–119 (2006).
    Article CAS Google Scholar
  17. Matta, H. & Chaudhary, P. M. Activation of alternative NF-κB pathway by human herpes virus 8-encoded Fas-associated death domain-like IL-1β-converting enzyme inhibitory protein (vFLIP). Proc. Natl Acad. Sci. USA 101, 9399–9404 (2004).
    Article CAS Google Scholar
  18. Matta, H., Mazzacurati, L., Schamus, S., Yang,.T, Sun, Q & Chaudhary, PM . Kaposi's sarcoma-associated herpesvirus (KSHV) oncoprotein K13 bypasses TRAFs and directly interacts with the IκB kinase complex to selectively activate NF-κB without JNK activation. J. Biol. Chem. 24, 24858–24865 (2007).
    Article Google Scholar
  19. Yang, J. K. et al. Crystal structure of MC159 reveals molecular mechanism of DISC assembly and FLIP inhibition. Mol. Cell 20, 939–949 (2005).
    Article CAS Google Scholar
  20. Yamada, Y. et al. The crystal structure of Atg3, an autophagy-related ubiquitin carrier protein (E2) enzyme that mediates Atg8 lipidation. J. Biol. Chem. 282, 8036–8043 (2007).
    Article CAS Google Scholar
  21. Bagneris, C. et al. Crystal structure of a vFlip–IKKγ complex: insights into viral activation of the IKK signalosome. Mol. Cell 30, 620–631 (2008).
    Article CAS Google Scholar
  22. Ye, F. C. et al. Kaposi's sarcoma-associated herpesvirus latent gene vFLIP inhibits viral lytic replication through NF-κB-mediated suppression of the AP-1 pathway: a novel mechanism of virus control of latency. J. Virol. 82, 4235–4249 (2008).
    Article CAS Google Scholar
  23. Gump, J. M. & Dowdy, S. F. TAT transduction: the molecular mechanism and therapeutic prospects. Trends Mol. Med. 13, 443–448 (2007).
    Article CAS Google Scholar
  24. Snyder, E. L., Meade, B. R., Saenz, C.C & Dowdy, S. F. Treatment of terminal peritoneal carcinomatosis by a transducible p53-activating peptide. PLos Biol. 2, 186–193 (2004).
    Article Google Scholar
  25. Wadia, J. S., Stan, R.V & Dowdy, S. F. Transducible TAT-HA fusogenic peptide enhances escape of TAT-fusion proteins after lipid raft macropinocytosis. Nature Med. 10, 310–315 (2004).
    Article CAS Google Scholar
  26. Hotchkiss, R. S. et al. TAT-BH4 and TAT-Bcl-xL peptides protect against sepsis-induced lymphocyte apoptosis in vivo. J. Immunol. 176, 5471–5477 (2006).
    Article CAS Google Scholar
  27. Wu, W., Rochford, R., Toomey, L., Harrington, W. Jr & Feuer, G. Inhibition of HHV-8/KSHV infected primary effusion lymphomas in NOD/SCID mice by azidothymidine and interferon-alpha. Leukemia Res. 29, 545–555 (2005).
    Article CAS Google Scholar
  28. Keller, S. A. et al. NF-kappaB is essential for the progression of KSHV- and EBV-infected lymphomas in vivo. Blood 107, 3295–3302 (2006).
    Article CAS Google Scholar
  29. Levine, B. Autophagy, immunity, and microbial adaptations. Cell Host Microb. 18, 527–549 (2009).
    Google Scholar
  30. Mizushima, N., Levine, B., Cuervo, A.M & Klionsky, D. J. Autophagy fights disease through cellular self-digestion. Nature 451, 1069–1075 (2008).
    Article CAS Google Scholar
  31. Maiuri, M. C., Zalckvar, E., Kimchi, A & Kroemer, G. Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nature Rev. Mol. Cell Biol. 8, 741–752 (2007).
    Article CAS Google Scholar
  32. Scott, R. C., Juhasz, G. & Neufeld, T. P. Direct induction of autophagy by Atg1 inhibits cell growth and induces apoptotic cell death. Curr. Biol. 17, 1–11 (2007).
    Article CAS Google Scholar
  33. Rubinztein, D. C., Gestwicki, J. E., Murphy, L.O & Klionsky, D. J. Potential therapeutic applications of autophagy. Nature Rev. Drug Discov. 6, 304–312 (2007).
    Article Google Scholar

Download references

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.

Author information

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

Authors

  1. Jong-Soo Lee
  2. Qinglin Li
  3. June-Yong Lee
  4. Sun-Hwa Lee
  5. Joseph H. Jeong
  6. Hye-Ra Lee
  7. Heesoon Chang
  8. Fu-Chun Zhou
  9. Shou-Jiang Gao
  10. Chengyu Liang
  11. Jae U. Jung

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.

Corresponding author

Correspondence toJae U. Jung.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Rights and permissions

About this article

Cite this article

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

Download citation

This article is cited by