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Bcl-2 complexed with Beclin-1 maintains full anti-apoptotic function

Oncogene volume 28, pages 2128–2141 (2009)Cite this article

Abstract

The binding of Bcl-2 to Beclin-1 reduces Beclin-1's capacity to induce autophagy. Here, we have tested whether the interaction is reciprocated by loss of Bcl-2's anti-apoptotic function. We targeted Bcl-2 to mitochondria or endoplasmic reticulum (ER) and induced apoptosis using several apoptotic stimuli that initiate ER and/or mitochondrial signaling pathways (UV radiation, TNF and cycloheximide, staurosporine, thapsigargin and tunicamycin). When Beclin-1 and Bcl-2 were expressed together in HeLa cells, Beclin-1 (but not Beclin-1 lacking the Bcl-2-binding domain) followed Bcl-2 to the appropriate organelle with complete or near-complete overlap (comprising 60 and 30% of cells, respectively). The interaction between Beclin-1 and Bcl-2 was verified by immunoprecipitation, and a membrane-proximate localization of Beclin-1 was shown by immunoelectron microscopy. Apoptosis was followed by measuring changes in nuclear morphology, caspase-3 activity, poly-ADP-ribose polymerase cleavage or punctation of mRFP-Bax on mitochondria. Binding of Beclin-1 to Bcl-2 did not modify apoptosis irrespective of Bcl-2 concentration, location or apoptotic stimulus. A similar result was obtained in Atg5−/− cells that are autophagy-deficient, arguing against compensation for the loss of protection by Bcl-2 by autophagy-mediated survival induced by Beclin-1. Hence, although Beclin-1 contains a BH3-only motif typical of pro-apoptotic proteins, it is a negligible modulator of Bcl-2's anti-apoptotic function.

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References

  • Adams JM, Cory S . (2007). The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene 26: 1324–1337.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Aita VM, Liang XH, Murty VV, Pincus DL, Yu W, Cayanis E et al. (1999). Cloning and genomic organization of beclin 1, a candidate tumor suppressor gene on chromosome 17q21. Genomics 59: 59–65.

    Article  CAS  PubMed  Google Scholar 

  • Akar U, Chaves-Reyez A, Barria M, Tari A, Sanguino A, Kondo Y et al. (2008). Silencing of Bcl-2 expression by small Interfering RNA induces autophagic cell death in MCF-7 breast cancer cells. Autophagy 4: 669–679.

    Article  CAS  PubMed  Google Scholar 

  • Annis MG, Yethon JA, Leber B, Andrews DW . (2004). There is more to life and death than mitochondria: Bcl-2 proteins at the endoplasmic reticulum. Biochim Biophys Acta 1644: 115–123.

    Article  CAS  PubMed  Google Scholar 

  • Arsov I, Li X, Matthews G, Coradin J, Hartmann B, Simon AK et al. (2008). BAC-mediated transgenic expression of fluorescent autophagic protein Beclin 1 reveals a role for Beclin 1 in lymphocyte development. Cell Death Differ 15: 1385–1395.

    Article  CAS  PubMed  Google Scholar 

  • Boya P, Gonzalez-Polo RA, Casares N, Perfettini JL, Dessen P, Larochette N et al. (2005). Inhibition of macroautophagy triggers apoptosis. Mol Cell Biol 25: 1025–1040.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cao Y, Klionsky DJ . (2007). Physiological functions of Atg6/Beclin 1: a unique autophagy-related protein. Cell Res 17: 839–849.

    Article  CAS  PubMed  Google Scholar 

  • Certo M, Del Gaizo Moore V, Nishino M, Wei G, Korsmeyer S, Armstrong SA et al. (2006). Mitochondria primed by death signals determine cellular addiction to antiapoptotic BCL-2 family members. Cancer Cell 9: 351–365.

    Article  CAS  PubMed  Google Scholar 

  • Chu CT, Zhu J, Dagda R . (2007). Beclin 1-independent pathway of damage-induced mitophagy and autophagic stress: implications for neurodegeneration and cell death. Autophagy 3: 663–666.

    Article  CAS  PubMed  Google Scholar 

  • Ciechomska IA, Goemans CG, Tolkovsky AM . (2008). Molecular links between autophagy and apoptosis. Methods Mol Biol 445: 175–193.

    Article  CAS  PubMed  Google Scholar 

  • Erlich S, Mizrachy L, Segev O, Lindenboim L, Zmira O, Adi-Harel S et al. (2007). Differential interactions between Beclin 1 and Bcl-2 family members. Autophagy 3: 561–568.

    Article  CAS  PubMed  Google Scholar 

  • Feng W, Huang S, Wu H, Zhang M . (2007). Molecular basis of Bcl-xL's target recognition versatility revealed by the structure of Bcl-xL in complex with the BH3 domain of Beclin-1. J Mol Biol 372: 223–235.

    Article  CAS  PubMed  Google Scholar 

  • Fimia GM, Stoykova A, Romagnoli A, Giunta L, Di Bartolomeo S, Nardacci R et al. (2007). Ambra1 regulates autophagy and development of the nervous system. Nature 447: 1121–1125.

    CAS  PubMed  Google Scholar 

  • Furuya N, Yu J, Byfield M, Pattingre S, Levine B . (2005). The evolutionarily conserved domain of Beclin 1 is required for Vps34 binding, autophagy and tumor suppressor function. Autophagy 1: 46–52.

    Article  CAS  PubMed  Google Scholar 

  • Goemans CG, Boya P, Skirrow CJ, Tolkovsky AM . (2008). Intra-mitochondrial degradation of Tim23 curtails the survival of cells rescued from apoptosis by caspase inhibitors. Cell Death Differ 15: 545–554.

    Article  CAS  PubMed  Google Scholar 

  • Hacki J, Egger L, Monney L, Conus S, Rosse T, Fellay I et al. (2000). Apoptotic crosstalk between the endoplasmic reticulum and mitochondria controlled by Bcl-2. Oncogene 19: 2286–2295.

    Article  CAS  PubMed  Google Scholar 

  • Hara T, Nakamura K, Matsui M, Yamamoto A, Nakahara Y, Suzuki-Migishima R et al. (2006). Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature 441: 885–889.

    Article  CAS  PubMed  Google Scholar 

  • Jin S, White E . (2007). Role of autophagy in cancer: management of metabolic stress. Autophagy 3: 28–31.

    Article  CAS  PubMed  Google Scholar 

  • Jin S, White E . (2008). Tumor suppression by autophagy through the management of metabolic stress. Autophagy 4: 563–566.

    Article  CAS  PubMed  Google Scholar 

  • Karantza-Wadsworth V, Patel S, Kravchuk O, Chen G, Mathew R, Jin S et al. (2007). Autophagy mitigates metabolic stress and genome damage in mammary tumorigenesis. Genes Dev 21: 1621–1635.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kaufmann T, Schlipf S, Sanz J, Neubert K, Stein R, Borner C . (2003). Characterization of the signal that directs Bcl-x(L), but not Bcl-2, to the mitochondrial outer membrane. J Cell Biol 160: 53–64.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Klionsky DJ . (2007). Autophagy: from phenomenology to molecular understanding in less than a decade. Nat Rev Mol Cell Biol 8: 931–937.

    Article  CAS  PubMed  Google Scholar 

  • Koike M, Shibata M, Tadakoshi M, Gotoh K, Komatsu M, Waguri S et al. (2008). Inhibition of autophagy prevents hippocampal pyramidal neuron death after hypoxic-ischemic injury. Am J Pathol 172: 454–469.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Komatsu M, Waguri S, Chiba T, Murata S, Iwata J, Tanida I et al. (2006). Loss of autophagy in the central nervous system causes neurodegeneration in mice. Nature 441: 880–884.

    Article  CAS  PubMed  Google Scholar 

  • Ku B, Woo JS, Liang C, Lee KH, Hong HS, E X et al. (2008). Structural and biochemical bases for the inhibition of autophagy and apoptosis by viral BCL-2 of murine gamma-herpesvirus 68. PLoS Pathog 4: e25.

    Article  PubMed  PubMed Central  Google Scholar 

  • Kuma A, Hatano M, Matsui M, Yamamoto A, Nakaya H, Yoshimori T et al. (2004). The role of autophagy during the early neonatal starvation period. Nature 432: 1032–1036.

    Article  CAS  PubMed  Google Scholar 

  • Levine B, Sinha S, Kroemer G . (2008). Bcl-2 family members: dual regulators of apoptosis and autophagy. Autophagy 4: 1–7.

    Article  Google Scholar 

  • Liang C, Feng P, Ku B, Dotan I, Canaani D, Oh BH et al. (2006). Autophagic and tumour suppressor activity of a novel Beclin1-binding protein UVRAG. Nat Cell Biol 8: 688–699.

    Article  CAS  PubMed  Google Scholar 

  • Liang XH, Jackson S, Seaman M, Brown K, Kempkes B, Hibshoosh H et al. (1999). Induction of autophagy and inhibition of tumorigenesis by beclin 1. Nature 402: 672–676.

    Article  CAS  PubMed  Google Scholar 

  • Liang XH, Kleeman LK, Jiang HH, Gordon G, Goldman JE, Berry G et al. (1998). Protection against fatal Sindbis virus encephalitis by beclin, a novel Bcl-2-interacting protein. J Virol 72: 8586–8596.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Maiuri MC, Criollo A, Tasdemir E, Vicencio JM, Tajeddine N, Hickman JA et al. (2007a). BH3-only proteins and BH3 mimetics induce autophagy by competitively disrupting the interaction between Beclin 1 and Bcl-2/Bcl-X(L). Autophagy 3: 374–376.

    Article  CAS  PubMed  Google Scholar 

  • Maiuri MC, Le Toumelin G, Criollo A, Rain JC, Gautier F, Juin P et al. (2007b). Functional and physical interaction between Bcl-X(L) and a BH3-like domain in Beclin-1. EMBO J 26: 2527–2539.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Maiuri MC, Zalckvar E, Kimchi A, Kroemer G . (2007c). Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol 8: 741–752.

    Article  CAS  PubMed  Google Scholar 

  • Meijer AJ, Codogno P . (2004). Regulation and role of autophagy in mammalian cells. Int J Biochem Cell Biol 36: 2445–2462.

    Article  CAS  PubMed  Google Scholar 

  • Michaelidis TM, Sendtner M, Cooper JD, Airaksinen MS, Holtmann B, Meyer M et al. (1996). Inactivation of Bcl-2 results in progressive degeneration of motoneurons, sympathetic and sensory neurons during early postnatal development. Neuron 17: 75–89.

    Article  CAS  PubMed  Google Scholar 

  • Mizushima N, Levine B, Cuervo AM, Klionsky DJ . (2008). Autophagy fights disease through cellular self-digestion. Nature 451: 1069–1075.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mizushima N, Ohsumi Y, Yoshimori T . (2002). Autophagosome formation in mammalian cells. Cell Struct Funct 27: 421–429.

    Article  PubMed  Google Scholar 

  • Noble CG, Dong JM, Manser E, Song H . (2008). BCL-XL and UVRAG cause a monomer-dimer switch in beclin1. J Biol Chem 283: 26274–26282.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Oberstein A, Jeffrey PD, Shi Y . (2007). Crystal structure of the Bcl-XL-Beclin 1 peptide complex: Beclin 1 is a novel BH3-only protein. J Biol Chem 282: 13123–13132.

    Article  CAS  PubMed  Google Scholar 

  • Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N et al. (2005). Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 122: 927–939.

    Article  CAS  PubMed  Google Scholar 

  • Qu X, Yu J, Bhagat G, Furuya N, Hibshoosh H, Troxel A et al. (2003). Promotion of tumorigenesis by heterozygous disruption of the beclin 1 autophagy gene. J Clin Invest 112: 1809–1820.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ravikumar B, Berger Z, Vacher C, O’Kane CJ, Rubinsztein DC . (2006). Rapamycin pre-treatment protects against apoptosis. Hum Mol Genet 15: 1209–1216.

    Article  CAS  PubMed  Google Scholar 

  • Samara C, Syntichaki P, Tavernarakis N . (2008). Autophagy is required for necrotic cell death in Caenorhabditis elegans. Cell Death Differ 15: 105–112.

    Article  CAS  PubMed  Google Scholar 

  • Scarlatti F, Maffei R, Beau I, Codogno P, Ghidoni R . (2008). Role of non-canonical Beclin 1-independent autophagy in cell death induced by resveratrol in human breast cancer cells. Cell Death Differ 15: 1318–1329.

    Article  CAS  PubMed  Google Scholar 

  • Scorrano L, Korsmeyer SJ . (2003). Mechanisms of cytochrome c release by proapoptotic BCL-2 family members. Biochem Biophys Res Commun 304: 437–444.

    Article  CAS  PubMed  Google Scholar 

  • Shimizu S, Kanaseki T, Mizushima N, Mizuta T, Arakawa-Kobayashi S, Thompson CB et al. (2004). Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes. Nat Cell Biol 6: 1221–1228.

    Article  CAS  PubMed  Google Scholar 

  • Sinha S, Colbert CL, Becker N, Wei Y, Levine B . (2008). Molecular basis of the regulation of Beclin 1-dependent autophagy by the gamma-herpesvirus 68 Bcl-2 homolog M11. Autophagy 4: 989–997.

    Article  CAS  PubMed  Google Scholar 

  • Skepper JN . (2000). Immunocytochemical strategies for electron microscopy: choice or compromise. J Microsc 199: 1–36.

    Article  CAS  PubMed  Google Scholar 

  • Swerdlow S, McColl K, Rong Y, Lam M, Gupta A, Distelhorst CW . (2008). Apoptosis inhibition by Bcl-2 gives way to autophagy in glucocorticoid-treated lymphocytes. Autophagy 4: 612–620.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Takahashi Y, Coppola D, Matsushita N, Cualing HD, Sun M, Sato Y et al. (2007). Bif-1 interacts with Beclin 1 through UVRAG and regulates autophagy and tumorigenesis. Nat Cell Biol 9: 1142–1151.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ullman E, Fan Y, Stawowczyk M, Chen HM, Yue Z, Zong WX . (2008). Autophagy promotes necrosis in apoptosis-deficient cells in response to ER stress. Cell Death Differ 15: 422–425.

    Article  CAS  PubMed  Google Scholar 

  • Yoshimori T . (2004). Autophagy: a regulated bulk degradation process inside cells. Biochem Biophys Res Commun 313: 453–458.

    Article  CAS  PubMed  Google Scholar 

  • Yue Z, Jin S, Yang C, Levine AJ, Heintz N . (2003). Beclin 1, an autophagy gene essential for early embryonic development, is a haploinsufficient tumor suppressor. Proc Natl Acad Sci USA 100: 15077–15082.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank our colleagues Beth Levine, Zenyu Yue, Noboru Mizushima, Said Aoufouchi, Christoph Borner, Goeff Butcher and Eiki Kominami for their generous gifts of reagents. This work was funded by program Grant no.064232 from the Wellcome Trust.

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Author notes

  1. G C Goemans

    Present address: 4Current address: Molecular Neurobiochemistry, Ruhr University Bochum, NC 7-171, D-44780 Bochum, Germany.,

Authors and Affiliations

  1. Department of Biochemistry, University of Cambridge, Cambridge, UK

    I A Ciechomska, G C Goemans & A M Tolkovsky

  2. Department of Cell Biology, Nencki Institute of Experimental Biology, Warsaw, Poland

    I A Ciechomska

  3. Department of Physiology, Development, and Neuroscience, University of Cambridge, Cambridge, UK

    J N Skepper

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  1. I A Ciechomska
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Correspondence to A M Tolkovsky.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Ciechomska, I., Goemans, G., Skepper, J. et al. Bcl-2 complexed with Beclin-1 maintains full anti-apoptotic function. Oncogene 28, 2128–2141 (2009). https://doi.org/10.1038/onc.2009.60

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