Apoptosis initiated by Bcl-2-regulated caspase activation independently of the cytochrome c/Apaf-1/caspase-9 apoptosome (original) (raw)

Nature volume 419, pages 634–637 (2002)Cite this article

Abstract

Apoptosis is an evolutionarily conserved cell suicide process executed by cysteine proteases (caspases) and regulated by the opposing factions of the Bcl-2 protein family1,2. Mammalian caspase-9 and its activator Apaf-1 were thought to be essential, because mice lacking either of them display neuronal hyperplasia and their lymphocytes and fibroblasts seem resistant to certain apoptotic stimuli3,4,5,6. Because Apaf-1 requires cytochrome c to activate caspase-9, and Bcl-2 prevents mitochondrial cytochrome c release, Bcl-2 is widely believed to inhibit apoptosis by safeguarding mitochondrial membrane integrity7,8,9. Our results suggest a different, broader role, because Bcl-2 overexpression increased lymphocyte numbers in mice and inhibited many apoptotic stimuli, but the absence of Apaf-1 or caspase-9 did not. Caspase activity was still discernible in cells lacking Apaf-1 or caspase-9, and a potent caspase antagonist both inhibited apoptosis and retarded cytochrome c release. We conclude that Bcl-2 regulates a caspase activation programme independently of the cytochrome c/Apaf-1/caspase-9 ‘apoptosome’, which seems to amplify rather than initiate the caspase cascade.

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

Access options

Subscribe to this journal

Receive 51 print issues and online access

$199.00 per year

only $3.90 per issue

Buy this article

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

Additional access options:

Similar content being viewed by others

References

  1. Hengartner, M. O. The biochemistry of apoptosis. Nature 407, 770–776 (2000)
    Article ADS CAS PubMed Google Scholar
  2. Cory, S. & Adams, J. M. The Bcl-2 family: Regulators of the cellular life-or-death switch. Nature Rev. Cancer 2, 647–656 (2002)
    Article CAS Google Scholar
  3. Kuida, K. et al. Reduced apoptosis and cytochrome c-mediated caspase activation in mice lacking caspase 9. Cell 94, 325–337 (1998)
    Article CAS PubMed Google Scholar
  4. Hakem, R. et al. Differential requirement for caspase 9 in apoptotic pathways in vivo. Cell 94, 339–352 (1998)
    Article CAS PubMed Google Scholar
  5. Yoshida, H. et al. Apaf1 is required for mitochondrial pathways of apoptosis and brain development. Cell 94, 739–750 (1998)
    Article CAS PubMed Google Scholar
  6. Cecconi, F., Alvarez-Bolado, G., Meyer, B. I., Roth, K. A. & Gruss, P. Apaf-1 (CED-4 homologue) regulates programmed cell death in mammalian development. Cell 94, 727–737 (1998)
    Article CAS PubMed Google Scholar
  7. Green, D. R. & Reed, J. C. Mitochondria and apoptosis. Science 281, 1309–1311 (1998)
    Article CAS PubMed Google Scholar
  8. Gross, A., McDonnell, J. M. & Korsmeyer, S. J. Bcl-2 family members and the mitochondria in apoptosis. Genes Dev. 13, 1899–1911 (1999)
    Article CAS PubMed Google Scholar
  9. Wang, X. The expanding role of mitochondria in apoptosis. Genes Dev. 15, 2922–2933 (2001)
    CAS PubMed Google Scholar
  10. Strasser, A., O'Connor, L. & Dixit, V. M. Apoptosis signaling. Annu. Rev. Biochem. 69, 217–245 (2000)
    Article CAS PubMed Google Scholar
  11. Hausmann, G. et al. Pro-apoptotic apoptosis protease-activating Factor 1 (Apaf-1) has a cytoplasmic localization distinct from Bcl-2 or Bcl-xL . J. Cell Biol. 149, 623–634 (2000)
    Article CAS PubMed PubMed Central Google Scholar
  12. Vaux, D. L., Weissman, I. L. & Kim, S. K. Prevention of programmed cell death in Caenorhabditis elegans by human _bcl_-2. Science 258, 1955–1957 (1992)
    Article ADS CAS PubMed Google Scholar
  13. Bouillet, P. et al. Proapoptotic Bcl-2 relative Bim required for certain apoptotic responses, leukocyte homeostasis, and to preclude autoimmunity. Science 286, 1735–1738 (1999)
    Article CAS PubMed Google Scholar
  14. Ogilvy, S. et al. Constitutive bcl-2 expression throughout the hematopoietic compartment affects multiple lineages and enhances progenitor cell survival. Proc. Natl Acad. Sci. USA 96, 14943–14948 (1999)
    Article ADS CAS PubMed PubMed Central Google Scholar
  15. Honarpour, N. et al. Adult Apaf-1-deficient mice exhibit male infertility. Dev. Biol. 218, 248–258 (2000)
    Article CAS PubMed Google Scholar
  16. Haraguchi, M. et al. Apoptotic protease activating factor 1 (Apaf-1)-independent cell death suppression by Bcl-2. J. Exp. Med. 191, 1709–1720 (2000)
    Article CAS PubMed PubMed Central Google Scholar
  17. Susin, S. A. et al. Two distinct pathways leading to nuclear apoptosis. J. Exp. Med. 192, 571–580 (2000)
    Article ADS CAS PubMed PubMed Central Google Scholar
  18. Liu, X., Zou, H., Slaughter, C. & Wang, X. DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 89, 175–184 (1997)
    Article CAS PubMed Google Scholar
  19. Enari, M. et al. A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391, 43–50 (1998)
    Article ADS CAS PubMed Google Scholar
  20. Wu, J. C. & Fritz, L. C. Irreversible caspase inhibitors: tools for studying apoptosis. Methods Enzymol. 17, 320–328 (1999)
    Article CAS Google Scholar
  21. Hoglen, N. C. et al. Characterization of the caspase inhibitor IDN-1965 in a model of apoptosis-associated liver injury. J. Pharmacol. Exp. Therapeut. 297, 811–818 (2001)
    CAS Google Scholar
  22. Nicholson, D. W. et al. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature 376, 37–43 (1995)
    Article ADS CAS PubMed Google Scholar
  23. Bouillet, P. et al. BH3-only Bcl-2 family member Bim is required for apoptosis of autoreactive thymocytes. Nature 415, 922–926 (2002)
    Article ADS CAS PubMed Google Scholar
  24. Hara, H. et al. The apoptotic protease-activating factor 1-mediated pathway of apoptosis is dispensable for negative selection of thymocytes. J. Immunol. 168, 2288–2295 (2002)
    Article CAS PubMed Google Scholar
  25. Lassus, P., Opitz-Araya, X. & Lazebnik, Y. Requirement for caspase-2 in stress-induced apoptosis before mitochondrial permeabilization. Science 297, 1352–1354 (2002)
    Article ADS CAS PubMed Google Scholar
  26. Dorstyn, L. et al. The role of cytochrome c in caspase activation in Drosophila melanogaster cells. J. Cell Biol. 156, 1089–1098 (2002)
    Article CAS PubMed PubMed Central Google Scholar
  27. Zimmermann, K. C., Ricci, J. E., Droin, N. M. & Green, D. R. The role of ARK in stress-induced apoptosis in Drosophila cells. J. Cell Biol. 156, 1077–1087 (2002)
    Article CAS PubMed PubMed Central Google Scholar
  28. Bergeron, L. et al. Defects in regulation of apoptosis in caspase-2-deficient mice. Genes Dev. 12, 1304–1314 (1998)
    Article CAS PubMed PubMed Central Google Scholar
  29. O'Reilly, L. A. et al. Caspase-2 is not required for thymocyte or neuronal apoptosis even though cleavage of caspase-2 is dependent on both Apaf-1 and caspase-9. Cell Death Differ. 9, 832–841 (2002)
    Article CAS PubMed Google Scholar
  30. Adams, J. M. & Cory, S. Apoptosomes: engines for caspase activation. Curr. Opin. Cell Biol. (in the press)

Download references

Acknowledgements

We thank Y. Lazebnik and P. Lassus for sharing unpublished results. We also thank P. Gruss for Apaf-1+/- mice, Y. Lazebnik and X. Opitz-Araya for monoclonal antibodies to caspases 3, 7 and 9, P. Vandenabeele and M. Kalai for the anti-mouse caspase-1 antibody, R. Anderson for the anti-HSP70 antibody and S. Nagata for the ICAD constructs. We thank E. Loza, A. Milligan, C. Tilbrook, A. Naughton and J. Merryful for animal care, F. Battye, D. Kaminaris, V. Lapatis, J. Chan and C. Tarlinton for cell sorting, S. Mifsud, L. DiRago, L.-C. Zhang and L. Tai for expert technical help and G. Filby for editorial assistance. We are grateful to S. Cory, A. Harris, K. Newton and H. Puthalakath for discussions and critical reading of this manuscript. This work was supported by fellowships and grants from the Dr Josef Steiner Cancer Research Foundation, the NHMRC, the Leukemia and Lymphoma Society (SCOR Center), the Anti-Cancer Council of Victoria, the Sylvia and Charles Viertel Charitable Foundation, the NIH, the AIRC and the Commonwealth Department of Education, Science and Training. F.C. is an Assistant Telethon Scientist.

Author information

Author notes

  1. Liam O'Connor
    Present address: Incyte Genomics, Palo Alto, California, 94304, USA
  2. Kevin J. Tomaselli, Jerry M. Adams and Andreas Strasser: These authors contributed equally to this work

Authors and Affiliations

  1. The Walter and Eliza Hall Institute, Melbourne, 3050, Australia
    Vanessa S. Marsden, Liam O'Connor, Lorraine A. O'Reilly, John Silke, Donald Metcalf, Paul G. Ekert, David C. S. Huang, David L. Vaux, Philippe Bouillet, Jerry M. Adams & Andreas Strasser
  2. Murdoch Children's Research Institute, Melbourne, 3050, Australia
    Paul G. Ekert
  3. Department of Biology, Universita Tor Vergata, Rome, Italy
    Francesco Cecconi
  4. Genomic Pharmacology, Vertex Pharmaceuticals, Cambridge, Massachusetts, 02139, USA
    Keisuke Kuida
  5. Idun Pharmaceuticals, San Diego, California, 92121, USA
    Kevin J. Tomaselli
  6. Merck-Frosst, Pointe-Claire-Dorval, H9H 3L1, Canada
    Sophie Roy & Don W. Nicholson

Authors

  1. Vanessa S. Marsden
    You can also search for this author inPubMed Google Scholar
  2. Liam O'Connor
    You can also search for this author inPubMed Google Scholar
  3. Lorraine A. O'Reilly
    You can also search for this author inPubMed Google Scholar
  4. John Silke
    You can also search for this author inPubMed Google Scholar
  5. Donald Metcalf
    You can also search for this author inPubMed Google Scholar
  6. Paul G. Ekert
    You can also search for this author inPubMed Google Scholar
  7. David C. S. Huang
    You can also search for this author inPubMed Google Scholar
  8. Francesco Cecconi
    You can also search for this author inPubMed Google Scholar
  9. Keisuke Kuida
    You can also search for this author inPubMed Google Scholar
  10. Kevin J. Tomaselli
    You can also search for this author inPubMed Google Scholar
  11. Sophie Roy
    You can also search for this author inPubMed Google Scholar
  12. Don W. Nicholson
    You can also search for this author inPubMed Google Scholar
  13. David L. Vaux
    You can also search for this author inPubMed Google Scholar
  14. Philippe Bouillet
    You can also search for this author inPubMed Google Scholar
  15. Jerry M. Adams
    You can also search for this author inPubMed Google Scholar
  16. Andreas Strasser
    You can also search for this author inPubMed Google Scholar

Corresponding author

Correspondence toAndreas Strasser.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

About this article

Cite this article

Marsden, V., O'Connor, L., O'Reilly, L. et al. Apoptosis initiated by Bcl-2-regulated caspase activation independently of the cytochrome c/Apaf-1/caspase-9 apoptosome.Nature 419, 634–637 (2002). https://doi.org/10.1038/nature01101

Download citation

This article is cited by