Activation and caspase-mediated inhibition of PARP A molecular switch between fibroblast necrosis and apoptosis in death receptor signaling

• Death ligands not only induce apoptosis but can also trigger necrosis with distinct biochemical and morphological features. We recently showed that in L929 cells CD95 ligation induces apoptosis, whereas TNF elicits necrosis. Treatment with anti-CD95 resulted in typical apoptosis characterized by caspase activation and DNA fragmentation. These events were barely induced by TNF, although TNF triggered cell death to a similar extent as CD95. Surprisingly, whereas the caspase inhibitor zVAD prevented CD95-mediated apoptosis, it potentiated TNF-induced necrosis. Cotreatment with TNF and zVAD was characterized by ATP depletion and accelerated necrosis. To investigate the mechanisms underlying TNF-induced cell death and its potentiation by zVAD, we examined the role of poly(ADP-ribose)polymerase-1 (PARP-1). TNF but not CD95 mediated PARP activation, whereas a PARP inhibitor suppressed TNF-induced necrosis and the sensitizing effect of zVAD. In addition, fibroblasts expressing a noncleavable PARP-I mutant were more sensitive to TNF than wild-tvpe cells. Our results indicate that TNF induces PARP activation leading to ATP depletion and subsequent necrosis. In contrast, in CD95-mediated apoptosis caspases cause PARP-1 cleavage and thereby maintain ATP levels. Because ATP is required for apoptosis, we suggest that PARP-1 cleavage functions as a molecular switch between apoptotic and necrotic modes of death receptor-induced cell death.

Subject headings

Natural Sciences  (hsv)

Biological Sciences  (hsv)

Cell Biology  (hsv)

Naturvetenskap  (hsv)

Biologiska vetenskaper  (hsv)

Cellbiologi  (hsv)

Biochemistry and Molecular Biology  (hsv)

Biokemi och molekylärbiologi  (hsv)

Keyword

cell-death

cleavage

damage

dna strand breaks

drug-induced apoptosis

gene

human poly(adp-ribose) polymerase

ischemia-reperfusion

l929 cells

Mice