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Analysis of candidate antagonists of IAP-mediated caspase inhibition using yeast reconstituted with the mammalian Apaf-1-activated apoptosis mechanism

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Abstract

We have reconstituted the Apaf-1-activated apoptosis mechanism in Sacchromyces cerevisiae such that the presence of a constitutively active form of Apaf-1 together with both Caspase-9 and Caspase-3 results in yeast death. This system is a good model of the Apaf-1-activated pathway in mammalian cells: MIHA (XIAP/hILP), and to a lesser degree MIHB (c-IAP1/HIAP2) and MIHC (c-IAP-2/HIAP1) can inhibit caspases in this system, and protection by IAPs (inhibitor of apoptosis) can be abrogated by coexpression of the Drosophila pro-apoptotic proteins HID and GRIM or the mammalian protein DIABLO/Smac. Using this system we demonstrate that unlike DIABLO/Smac, other proteins which interact with mammalian IAPs (TAB-1, Zap-1, Traf-1 and Traf-2) do not act to antagonise IAP- mediated caspase inhibition.

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Authors and Affiliations

  1. Department of Haematology and Oncology, Murdoch Children's Research Institute, Australia

    C. J. Hawkins, R. Foster & D. M. Ashley

  2. The Walter and Eliza Hall Institute of Medical Research, Post Office Royal Melbourne Hospital, Parkville, 3050, Australia

    J. Silke, A. M. Verhagen & P. G. Ekert

  3. Department of Neonatology, Royal Children's Hospital, Flemington Road, Parkville, 3052, Australia

    P. G. Ekert

  4. Department of Paediatrics, University of Melbourne, Parkville, 3010, Australia

    C. J. Hawkins

  5. Department of Paediatrics, University of Melbourne, Parkville, 3010, Australia

    R. Foster & D. M. Ashley

Authors
  1. C. J. Hawkins
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  2. J. Silke
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  3. A. M. Verhagen
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  4. R. Foster
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  5. P. G. Ekert
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  6. D. M. Ashley
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Hawkins, C.J., Silke, J., Verhagen, A.M. et al. Analysis of candidate antagonists of IAP-mediated caspase inhibition using yeast reconstituted with the mammalian Apaf-1-activated apoptosis mechanism. Apoptosis 6, 331–338 (2001). https://doi.org/10.1023/A:1011329917895

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