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. 1997 Apr 1;99(7):1751–1758. doi: 10.1172/JCI119339

Activation of CPP32-like protease in tumor necrosis factor-induced apoptosis is dependent on mitochondrial function.

M Higuchi 1, B B Aggarwal 1, E T Yeh 1
PMCID: PMC507996  PMID: 9120020

Abstract

Mitochondria have been implicated in apoptosis, however, the precise mechanisms whereby mitochondria exert their effect are not clear. To gain further insights, we generated a panel of cells from ML-1a cells that were rendered respiration deficient by ethidium bromide treatment. Two respiration-deficient clones were subsequently reconstituted by fusion with platelets. Respiration-deficient clones were resistant to TNF-induced apoptosis, whereas ML-1a and reconstituted clones were sensitive. In contrast, inhibition of proliferation and induction of differentiation by TNF were still observed in respiration deficient clones, suggesting a selective requirement of respiration in TNF-induced apoptosis. Furthermore the apoptosis machinery is not completely altered in respiration-deficient cells because they underwent apoptosis after staurosporine treatment. Next, we showed that apoptosis induced by TNF and staurosporine were blocked by z-DEVD-CH2F, an inhibitor of CPP32-like cysteine protease, suggesting the involvement of CPP32-like protease in both apoptosis signaling pathways. Interestingly, TNF activated CPP32-like protease in the parental and reconstituted clones but not in respiration-deficient clones, and staurosporine in all clones. Thus, the apoptosis signaling block in respiration-deficient clones is located at a step before CPP32-like protease activation, which can be bypassed by staurosporine.

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