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p53 accumulation in apoptotic macrophages is an energy demanding process that precedes cytochrome c release in response to nitric oxide

Oncogene volume 18, pages 6403–6410 (1999)Cite this article

Abstract

Apoptosis in response to stress signals activates effector caspases known to be regulated by the release of cytochrome c (Cyt c) from mitochondria and the subsequent ATP-dependent activation of the death regulator apoptotic protease-activating factor 1 (Apaf-1). Experiments were carried out to determine whether the release of Cyt c is evoked by NO□ in RAW 264.7 macrophages and to position signaling components relative to mitochondria. S-nitrosoglutathione and spermine-NO caused a fast p53 accumulation, followed by Bcl-xL downregulation, Cyt c release, and caspase activation. These alterations were absent in p53 antisense expressing macrophages (RΔp53asn-11). In Bcl-2 overexpressing cells (Rbcl2-14) Cyt c relocation and caspase activation were abrogated although p53 accumulation remained intact. The use of caspase inhibitors revealed Cyt c release and decreased Bcl-xL expression to be caspase independent. ATP-depleted cells showed a shift from apoptosis towards necrosis and no p53 accumulation or caspase activation upon NO□ addition. Conclusively, NO□-mediated apoptosis in macrophages is entirely controlled by the mitochondrial pathway with the implication that Cyt c relocation demands p53 accumulation. Moreover, pulse-chase-experiments in combination with the ATP-depletion protocol identified p53 accumulation and stabilization as an energy requiring process. This allowed to dissect two ATP-dependent steps, one is in association with Apaf-1 formation, while the other resides in p53 accumulation.

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Acknowledgements

We thank Brigitte Rogge for excellent technical assistance. This work was supported by the Deutsche Forschungsgemeinschaft and the Deutsche Krebshilfe.

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  1. Department of Medicine IV-Experimental Division, University of Erlangen-Nürnberg, Faculty of Medicine, Loschgestrasse 8, 91054, Erlangen, Germany

    Florian Brockhaus & Bernhard Brüne

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Brockhaus, F., Brüne, B. p53 accumulation in apoptotic macrophages is an energy demanding process that precedes cytochrome c release in response to nitric oxide. Oncogene 18, 6403–6410 (1999). https://doi.org/10.1038/sj.onc.1203058

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