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. 1997 Dec 15;16(24):7372–7381. doi: 10.1093/emboj/16.24.7372

Reaper-induced apoptosis in a vertebrate system.

E K Evans 1, T Kuwana 1, S L Strum 1, J J Smith 1, D D Newmeyer 1, S Kornbluth 1
PMCID: PMC1170337  PMID: 9405366

Abstract

The reaper protein of Drosophila melanogaster has been shown to be a central regulator of apoptosis in that organism. However, it has not been shown to function in any vertebrate nor have the cellular components required for its action been defined. In this report we show that reaper can induce rapid apoptosis in vitro using an apoptotic reconstitution system derived from Xenopus eggs. Moreover, we show that a subcellular fraction enriched in mitochondria is required for this process and that reaper, acting in conjunction with cytosolic factors, can trigger mitochondrial cytochrome c release. Bcl-2 antagonizes these effects, but high levels of reaper can overcome the Bcl-2 block. These results demonstrate that reaper can function in a vertebrate context, suggesting that reaper-responsive factors are conserved elements of the apoptotic program.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bellamy C. O., Malcomson R. D., Harrison D. J., Wyllie A. H. Cell death in health and disease: the biology and regulation of apoptosis. Semin Cancer Biol. 1995 Feb;6(1):3–16. doi: 10.1006/scbi.1995.0002. [DOI] [PubMed] [Google Scholar]
  2. Boldin M. P., Goncharov T. M., Goltsev Y. V., Wallach D. Involvement of MACH, a novel MORT1/FADD-interacting protease, in Fas/APO-1- and TNF receptor-induced cell death. Cell. 1996 Jun 14;85(6):803–815. doi: 10.1016/s0092-8674(00)81265-9. [DOI] [PubMed] [Google Scholar]
  3. Bump N. J., Hackett M., Hugunin M., Seshagiri S., Brady K., Chen P., Ferenz C., Franklin S., Ghayur T., Li P. Inhibition of ICE family proteases by baculovirus antiapoptotic protein p35. Science. 1995 Sep 29;269(5232):1885–1888. doi: 10.1126/science.7569933. [DOI] [PubMed] [Google Scholar]
  4. Chen P., Lee P., Otto L., Abrams J. Apoptotic activity of REAPER is distinct from signaling by the tumor necrosis factor receptor 1 death domain. J Biol Chem. 1996 Oct 18;271(42):25735–25737. doi: 10.1074/jbc.271.42.25735. [DOI] [PubMed] [Google Scholar]
  5. Chinnaiyan A. M., Dixit V. M. The cell-death machine. Curr Biol. 1996 May 1;6(5):555–562. doi: 10.1016/s0960-9822(02)00541-9. [DOI] [PubMed] [Google Scholar]
  6. Chinnaiyan A. M., O'Rourke K., Lane B. R., Dixit V. M. Interaction of CED-4 with CED-3 and CED-9: a molecular framework for cell death. Science. 1997 Feb 21;275(5303):1122–1126. doi: 10.1126/science.275.5303.1122. [DOI] [PubMed] [Google Scholar]
  7. Chinnaiyan A. M., O'Rourke K., Tewari M., Dixit V. M. FADD, a novel death domain-containing protein, interacts with the death domain of Fas and initiates apoptosis. Cell. 1995 May 19;81(4):505–512. doi: 10.1016/0092-8674(95)90071-3. [DOI] [PubMed] [Google Scholar]
  8. Duan H., Chinnaiyan A. M., Hudson P. L., Wing J. P., He W. W., Dixit V. M. ICE-LAP3, a novel mammalian homologue of the Caenorhabditis elegans cell death protein Ced-3 is activated during Fas- and tumor necrosis factor-induced apoptosis. J Biol Chem. 1996 Jan 19;271(3):1621–1625. doi: 10.1074/jbc.271.3.1621. [DOI] [PubMed] [Google Scholar]
  9. Duan H., Dixit V. M. RAIDD is a new 'death' adaptor molecule. Nature. 1997 Jan 2;385(6611):86–89. doi: 10.1038/385086a0. [DOI] [PubMed] [Google Scholar]
  10. Evans E. K., Lu W., Strum S. L., Mayer B. J., Kornbluth S. Crk is required for apoptosis in Xenopus egg extracts. EMBO J. 1997 Jan 15;16(2):230–241. doi: 10.1093/emboj/16.2.230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Farschon D. M., Couture C., Mustelin T., Newmeyer D. D. Temporal phases in apoptosis defined by the actions of Src homology 2 domains, ceramide, Bcl-2, interleukin-1beta converting enzyme family proteases, and a dense membrane fraction. J Cell Biol. 1997 Jun 2;137(5):1117–1125. doi: 10.1083/jcb.137.5.1117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Golstein P., Marguet D., Depraetere V. Fas bridging cell death and cytotoxicity: the reaper connection. Immunol Rev. 1995 Aug;146:45–56. doi: 10.1111/j.1600-065x.1995.tb00683.x. [DOI] [PubMed] [Google Scholar]
  13. Hsu H., Xiong J., Goeddel D. V. The TNF receptor 1-associated protein TRADD signals cell death and NF-kappa B activation. Cell. 1995 May 19;81(4):495–504. doi: 10.1016/0092-8674(95)90070-5. [DOI] [PubMed] [Google Scholar]
  14. Kluck R. M., Bossy-Wetzel E., Green D. R., Newmeyer D. D. The release of cytochrome c from mitochondria: a primary site for Bcl-2 regulation of apoptosis. Science. 1997 Feb 21;275(5303):1132–1136. doi: 10.1126/science.275.5303.1132. [DOI] [PubMed] [Google Scholar]
  15. Kluck R. M., Martin S. J., Hoffman B. M., Zhou J. S., Green D. R., Newmeyer D. D. Cytochrome c activation of CPP32-like proteolysis plays a critical role in a Xenopus cell-free apoptosis system. EMBO J. 1997 Aug 1;16(15):4639–4649. doi: 10.1093/emboj/16.15.4639. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lazebnik Y. A., Kaufmann S. H., Desnoyers S., Poirier G. G., Earnshaw W. C. Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature. 1994 Sep 22;371(6495):346–347. doi: 10.1038/371346a0. [DOI] [PubMed] [Google Scholar]
  17. Liu X., Kim C. N., Yang J., Jemmerson R., Wang X. Induction of apoptotic program in cell-free extracts: requirement for dATP and cytochrome c. Cell. 1996 Jul 12;86(1):147–157. doi: 10.1016/s0092-8674(00)80085-9. [DOI] [PubMed] [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. 1997 Apr 18;89(2):175–184. doi: 10.1016/s0092-8674(00)80197-x. [DOI] [PubMed] [Google Scholar]
  19. Muzio M., Chinnaiyan A. M., Kischkel F. C., O'Rourke K., Shevchenko A., Ni J., Scaffidi C., Bretz J. D., Zhang M., Gentz R. FLICE, a novel FADD-homologous ICE/CED-3-like protease, is recruited to the CD95 (Fas/APO-1) death--inducing signaling complex. Cell. 1996 Jun 14;85(6):817–827. doi: 10.1016/s0092-8674(00)81266-0. [DOI] [PubMed] [Google Scholar]
  20. Newmeyer D. D., Farschon D. M., Reed J. C. Cell-free apoptosis in Xenopus egg extracts: inhibition by Bcl-2 and requirement for an organelle fraction enriched in mitochondria. Cell. 1994 Oct 21;79(2):353–364. doi: 10.1016/0092-8674(94)90203-8. [DOI] [PubMed] [Google Scholar]
  21. Nicholson D. W., Ali A., Thornberry N. A., Vaillancourt J. P., Ding C. K., Gallant M., Gareau Y., Griffin P. R., Labelle M., Lazebnik Y. A. Identification and inhibition of the ICE/CED-3 protease necessary for mammalian apoptosis. Nature. 1995 Jul 6;376(6535):37–43. doi: 10.1038/376037a0. [DOI] [PubMed] [Google Scholar]
  22. Nuñez G., Clarke M. F. The Bcl-2 family of proteins: regulators of cell death and survival. Trends Cell Biol. 1994 Nov;4(11):399–403. doi: 10.1016/0962-8924(94)90053-1. [DOI] [PubMed] [Google Scholar]
  23. Pronk G. J., Ramer K., Amiri P., Williams L. T. Requirement of an ICE-like protease for induction of apoptosis and ceramide generation by REAPER. Science. 1996 Feb 9;271(5250):808–810. doi: 10.1126/science.271.5250.808. [DOI] [PubMed] [Google Scholar]
  24. Reed J. C. Bcl-2 and the regulation of programmed cell death. J Cell Biol. 1994 Jan;124(1-2):1–6. doi: 10.1083/jcb.124.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Spector M. S., Desnoyers S., Hoeppner D. J., Hengartner M. O. Interaction between the C. elegans cell-death regulators CED-9 and CED-4. Nature. 1997 Feb 13;385(6617):653–656. doi: 10.1038/385653a0. [DOI] [PubMed] [Google Scholar]
  26. Tewari M., Quan L. T., O'Rourke K., Desnoyers S., Zeng Z., Beidler D. R., Poirier G. G., Salvesen G. S., Dixit V. M. Yama/CPP32 beta, a mammalian homolog of CED-3, is a CrmA-inhibitable protease that cleaves the death substrate poly(ADP-ribose) polymerase. Cell. 1995 Jun 2;81(5):801–809. doi: 10.1016/0092-8674(95)90541-3. [DOI] [PubMed] [Google Scholar]
  27. Vucic D., Seshagiri S., Miller L. K. Characterization of reaper- and FADD-induced apoptosis in a lepidopteran cell line. Mol Cell Biol. 1997 Feb;17(2):667–676. doi: 10.1128/mcb.17.2.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. White K., Grether M. E., Abrams J. M., Young L., Farrell K., Steller H. Genetic control of programmed cell death in Drosophila. Science. 1994 Apr 29;264(5159):677–683. doi: 10.1126/science.8171319. [DOI] [PubMed] [Google Scholar]
  29. White K., Tahaoglu E., Steller H. Cell killing by the Drosophila gene reaper. Science. 1996 Feb 9;271(5250):805–807. doi: 10.1126/science.271.5250.805. [DOI] [PubMed] [Google Scholar]
  30. Xue D., Horvitz H. R. Inhibition of the Caenorhabditis elegans cell-death protease CED-3 by a CED-3 cleavage site in baculovirus p35 protein. Nature. 1995 Sep 21;377(6546):248–251. doi: 10.1038/377248a0. [DOI] [PubMed] [Google Scholar]
  31. Yang J., Liu X., Bhalla K., Kim C. N., Ibrado A. M., Cai J., Peng T. I., Jones D. P., Wang X. Prevention of apoptosis by Bcl-2: release of cytochrome c from mitochondria blocked. Science. 1997 Feb 21;275(5303):1129–1132. doi: 10.1126/science.275.5303.1129. [DOI] [PubMed] [Google Scholar]
  32. Yuan J. Molecular control of life and death. Curr Opin Cell Biol. 1995 Apr;7(2):211–214. doi: 10.1016/0955-0674(95)80030-1. [DOI] [PubMed] [Google Scholar]

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