Abstract
The apoptosis-associated speck-like protein (ASC) is an unusual adaptor protein that contains the Pyrin/PAAD death domain in addition to the CARD protein–protein interaction domain1,2,3,4,5. Here, we present evidence that ASC can function as an adaptor molecule for Bax and regulate a p53–Bax mitochondrial pathway of apoptosis. When ectopically expressed, ASC interacted directly with Bax, colocalized with Bax to the mitochondria, induced cytochrome c release with a significant reduction of mitochondrial membrane potential and resulted in the activation of caspase-9, -2 and -3. The rapid induction of apoptosis by ASC was not observed in Bax-deficient cells. We also show that induction of ASC after exposure to genotoxic stress is dependent on p53. Blocking of endogenous ASC expression by small-interfering RNA (siRNA) reduced the apoptotic response and inhibited translocation of Bax to mitochondria in response to p53 or genotoxic insult, suggesting that ASC is required to translocate Bax to the mitochondria. Our findings demonstrate that ASC has an essential role in the intrinsic mitochondrial pathway of apoptosis through a p53–Bax network.
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Acknowledgements
We thank R. Brent, P.M. Vertino and K. Nakayama for providing plasmids, Myc-tagged caspase-2, Myc-tagged ASC and Bcl2−/− mouse embryonic fibroblasts, respectively. We thank Z. Ronai, J. Menfredi, D. Sasson and J. Licht for helpful discussion, J. Kwak and P. Ongusaha for assistance and critical reading, and M. Shirane for technical advice and the HA-Bax plasmid. We are grateful to Y. Lazebnik for providing us with IMR90-E1A and several plasmid constructs of wild-type and mutant caspase-2, and B. Vogelstein for providing the HCT116 Bax-null cell line. This work was supported by National Institutes of Health grants CA78356, CA82211 and CA80058.
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Ohtsuka, T., Ryu, H., Minamishima, Y. et al. ASC is a Bax adaptor and regulates the p53–Bax mitochondrial apoptosis pathway. Nat Cell Biol 6, 121–128 (2004). https://doi.org/10.1038/ncb1087
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DOI: https://doi.org/10.1038/ncb1087
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