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Published March 18, 2002 | Published
Journal Article Open

The role of cytochrome c in caspase activation in Drosophila melanogaster cells

Abstract

The release of cytochrome c from mitochondria is necessary for the formation of the Apaf-1 apoptosome and subsequent activation of caspase-9 in mammalian cells. However, the role of cytochrome c in caspase activation in Drosophila cells is not well understood. We demonstrate here that cytochrome c remains associated with mitochondria during apoptosis of Drosophila cells and that the initiator caspase DRONC and effector caspase DRICE are activated after various death stimuli without any significant release of cytochrome c in the cytosol. Ectopic expression of the proapoptotic Bcl-2 protein, DEBCL, also fails to show any cytochrome c release from mitochondria. A significant proportion of cellular DRONC and DRICE appears to localize near mitochondria, suggesting that an apoptosome may form in the vicinity of mitochondria in the absence of cytochrome c release. In vitro, DRONC was recruited to a >700-kD complex, similar to the mammalian apoptosome in cell extracts supplemented with cytochrome c and dATP. These results suggest that caspase activation in insects follows a more primitive mechanism that may be the precursor to the caspase activation pathways in mammals.

Additional Information

© 2002 The Rockefeller University Press. Submitted: 29 November 2001; revised: 1 February 2002; accepted: 4 February 2002. We are grateful to Dr. K. Ui-Tei for BG2 cells, Dr. A. Dorn for l(2)mbn cells, and members of our laboratory for sharing ideas and reagents. This work was supported by a grant and a fellowship to S. Kumar from the National Health and Medical Research Council of Australia.

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