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J Biol Chem, Vol. 273, Issue 25, 15540-15545, June 19, 1998

Caspase-3 Is Required for alpha -Fodrin Cleavage but Dispensable for Cleavage of Other Death Substrates in Apoptosis

Reiner U. Jänicke, Patrick Ng, Michael L. Sprengart, and Alan G. Porter

From the Institute of Molecular and Cell Biology, The National University of Singapore, 30 Medical Dr., Singapore 117609, Republic of Singapore

Although the commonly activated death protease caspase-3 appears not to be essential for apoptosis during development except in the brain, it was not shown whether substrates known to be cleaved by caspase-3 are still proteolyzed in its absence. We have addressed this question with MCF-7 breast carcinoma cells that we recently showed lack caspase-3 owing to the functional deletion of the CASP-3 gene. Tumor necrosis factor- or staurosporine-induced apoptosis of caspase-3-deficient MCF-7 cells resulted in cleavage of the death substrates PARP, Rb, PAK2, DNA-PKcs, gelsolin, and DFF-45, but not alpha -fodrin. In contrast, all these substrates including alpha -fodrin were cleaved in apoptotic HeLa cells expressing caspase-3. Introduction of CASP-3 cDNA, but not CASP-10 cDNA, into MCF-7 cells restored alpha -fodrin cleavage. In addition, tumor necrosis factor- or staurosporine-induced apoptosis of MCF-7 cells stably expressing pro-caspase-3 also resulted in alpha -fodrin cleavage. Although the specific caspase inhibitory peptides Z-VAD-fmk and Z-DEVD-fmk prevented apoptosis of MCF-7 cells, we were unable to detect activation of caspases 2 and 7, which are known to be inhibited by Z-DEVD-fmk. Together our results suggest that caspase-3 is essential for cleavage of alpha -fodrin, but dispensable for the cleavage of PARP, Rb, PAK2, DNA-PKcs, gelsolin, and DFF-45 and imply that one or more caspases other than caspases 2, 3, and 7 is activated and plays a crucial role in the cleavage of these substrates in MCF-7 cells.


Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.



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