D. Mcilroy et al., Involvement of caspase 3-activated DNase in internucleosomal DNA cleavage induced by diverse apoptotic stimuli, ONCOGENE, 18(31), 1999, pp. 4401-4408
Degradation of chromosomal DNA into nucleosome-sized fragments is one of th
e characteristics of apoptotic cell death. Here, we examined whether caspas
e-activated DNase (CAD) is responsible for the DNA fragmentation that occur
s upon exposure to various apoptotic stimuli. When human Jurkat cells were
exposed to etoposide, or UV or gamma radiation, a caspase-3-like protease w
as activated, and nuclear DNA was fragmented. Human TF-1 cells, which are d
ependent on granulocyte-macrophage colony-stimulating factor (GM-CSF), also
underwent apoptosis accompanied by the activation of caspase-3-like protea
se and DNA fragmentation, when cultured without the cytokine. Both Jurkat a
nd TF-1 cells expressed two forms of ICAD, ICAD-L and ICAD-S, which were cl
eaved upon exposure to these apoptotic stimuli. Among eight different caspa
ses examined, recombinant caspases 3 and 7 specifically cleaved ICAD synthe
sized in a cell-free system. An expression plasmid containing mouse ICAD-L
mutated at the caspase-3-recognition sites was then introduced into Jurkat
and TF-1 cells. When the transformants were induced to undergo apoptosis (b
y treatment with etoposide, UV or gamma radiation for Jurkat cells, or fact
or withdrawal for TF-1 cells) they did not show DNA fragmentation, although
they still died as a result of these stimuli. These results indicated that
CAD, released from ICAD by caspase activation, is involved in the nuclear
DNA fragmentation induced by these apoptotic stimuli.