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J Biol Chem, Vol. 273, Issue 25, 15540-15545, June 19, 1998
-Fodrin Cleavage but Dispensable for
Cleavage of Other Death Substrates in Apoptosis
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
-fodrin. In
contrast, all these substrates including
-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
-fodrin cleavage. In addition, tumor necrosis factor- or staurosporine-induced apoptosis of MCF-7 cells stably expressing pro-caspase-3 also resulted in
-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
-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.
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