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A more recent version of this article appeared on March 1, 2002
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Submitted on May 30, 2001
Revised on November 1, 2001
Accepted on November 26, 2001
1 Department of Immunology and Cell Biology, University of Münster, Röntgenstr. 21, D-48149 Münster, Germany
2 Department of Immunology and Cell Biology, University of Münster, Münster, Germany
3 Department of Experimental and Diagnostic Medicine, University of Ferrara, Ferrara, Italy
4 International Agency for Research on Cancer (IARC), Lyon, France
* Corresponding author. E-mail address: los{at}uni-muenster.de.
Death ligands do 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 non-cleavable PARP-1 mutant were more sensitive to TNF than wild-type 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. As 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.
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