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SMAC/Diablo mediates the proapoptotic function of PUMA by regulating PUMA-induced mitochondrial events

Oncogene volume 26, pages 4189–4198 (2007)Cite this article

Abstract

p53-upregulated modulator of apoptosis (PUMA) is a BH3-only Bcl-2 family protein and an essential mediator of DNA damage-induced apoptosis. PUMA is localized in the mitochondria and induces apoptosis through the mitochondrial pathway. However, the mechanisms of PUMA-induced apoptosis remain unclear. In this study, we found that second mitochondria-derived activator of caspase (SMAC)/Diablo, a mitochondrial apoptogenic protein, mediates the proapoptotic function of PUMA by regulating PUMA-induced mitochondrial events. SMAC is consistently released into the cytosol in colon cancer cells undergoing PUMA-induced apoptosis. In SMAC-deficient cells, execution of PUMA-induced apoptosis is abrogated, in company with decreases in caspase activation, cytosolic release of cytochrome c and collapse of mitochondrial membrane potential. Reconstituting SMAC expression restored these events in the SMAC-deficient cells. Furthermore, SMAC and agents that mimic the inhibitor of apoptosis proteins (IAPs) inhibition function of SMAC significantly sensitize cells to PUMA-induced apoptosis. These results demonstrate an important role of SMAC in executing DNA damage-induced and PUMA-mediated apoptosis and suggest that SMAC participates in a feedback amplification loop to promote cytochrome c release and other mitochondrial events in apoptosis.

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Acknowledgements

We thank Dr Xiaodong Wang and Dr Patrick G Harran at the University of Texas Southwestern Medical Center for providing the SMAC mimetic and control compounds, Dr Eileen White for providing the SMAC antibody, Dr Chunying Du at the Stowers Institute for Medical Research for providing the SMAC expression plasmid, Ms Hongtao Liu for technical assistance and members of our laboratories for helpful discussions. This work is supported by NIH Grant CA106348, grants from the Edward Mallinckrodt Jr Foundation and Elsa U Pardee foundation (LZ) and grants from the Flight Attendant Medical Research Institute (FAMRI), the Alliance for Cancer Gene Therapy (ACGT) and the Hillman Foundation (to JY). LZ is a scholar of the General Motors (GM) Cancer Research Foundation and the V Foundation for Cancer Research.

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  1. Departments of Pharmacology and Pathology, University of Pittsburgh Cancer Institute, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    J Yu, P Wang, L Ming, M A Wood & L Zhang

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  1. J Yu
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Correspondence to L Zhang.

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Yu, J., Wang, P., Ming, L. et al. SMAC/Diablo mediates the proapoptotic function of PUMA by regulating PUMA-induced mitochondrial events. Oncogene 26, 4189–4198 (2007). https://doi.org/10.1038/sj.onc.1210196

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