Abstract
Mitochondrial proteins function as essential regulators in apoptosis. Here, we show that mitochondrial adenylate kinase 2 (AK2) mediates mitochondrial apoptosis through the formation of an AK2–FADD–caspase-10 (AFAC10) complex. Downregulation of AK2 attenuates etoposide- or staurosporine-induced apoptosis in human cells, but not that induced by tumour-necrosis-factor-related apoptosis-inducing ligand (TRAIL) or Fas ligand (FasL). During intrinsic apoptosis, AK2 translocates to the cytoplasm, whereas this event is diminished in Apaf-1 knockdown cells and prevented by Bcl-2 or Bcl-XL. Addition of purified AK2 protein to cell extracts first induces activation of caspase-10 via FADD and subsequently caspase-3 activation, but does not affect caspase-8. AFAC10 complexes are detected in cells undergoing intrinsic cell death and AK2 promotes the association of caspase-10 with FADD. In contrast, AFAC10 complexes are not detected in several etoposide-resistant human tumour cell lines. Taken together, these results suggest that, acting in concert with FADD and caspase-10, AK2 mediates a novel intrinsic apoptotic pathway that may be involved in tumorigenesis.
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Acknowledgements
The authors thank: J. Blenis (Harvard University, Boston, MA) for providing Jurkat cells (A3, I2-1, I9-2); J. Yuan (Harvard University) for HeLa/Bcl-2, HeLa/Bcl-XL and HeLa/CrmA cells; and H. D. Ryoo (New York University, New York, NY) for critical reading of this manuscript. J.-O. P. and S.-h. H. were in part supported by the Korean Research Foundation (KRF-2005-201-C00039) and by the BK21 programme. This work was supported by grants from the Functional Human Genome Project (FG06-2-7) and the Brain Research Center of the 21st Century Frontier Research Programme and the Ubiquitome project funded by the Korea Science and Engineering Foundation (KOSEF) of the Korea government (MOST).
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H.J.L., J.O.P. and H.J.K. performed flow cytometry analysis, IP assays and cell line characterization. H.J.L., Y.O., S.H.H., H.K., D.H.C., H.N.W. and J.H.N. performed siRNA generation and fractionation and RT–PCR analysis. H.J.L. and Y.J.J. performed in vitro pull-down assays. S.S. performed functional assays of AK2 and FADD in the mouse system. Hyo Joon Kim and K.S.K. performed antibody generation and mutagenesis. Y.K.J. guided the project and data interpretation. H.J.L. and Y.K.J. wrote the manuscript.
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Lee, HJ., Pyo, JO., Oh, Y. et al. AK2 activates a novel apoptotic pathway through formation of a complex with FADD and caspase-10. Nat Cell Biol 9, 1303–1310 (2007). https://doi.org/10.1038/ncb1650
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DOI: https://doi.org/10.1038/ncb1650
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