Caspase-2 resides in the mitochondria and mediates apoptosis directly from the mitochondrial compartment

M. Lopez-Cruzan, R. Sharma, M. Tiwari, S. Karbach, D. Holstein, C. R. Martin, J. D. Lechleiter, B. Herman

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Caspase-2 plays an important role in apoptosis induced by several stimuli, including oxidative stress. However, the subcellular localization of caspase-2, particularly its presence in the mitochondria, is unclear. It is also not known if cytosolic caspase-2 translocates to the mitochondria to trigger the intrinsic pathway of apoptosis or if caspase-2 is constitutively present in the mitochondria that then selectively mediates this apoptotic effect. Here, we demonstrate the presence of caspase-2 in purified mitochondrial fractions from in vitro-cultured cells and in liver hepatocytes using immunoblots and confocal microscopy. We show that mitochondrial caspase-2 is functionally active by performing fluorescence resonance energy transfer analyses using a mitochondrially targeted substrate flanked by donor and acceptor fluorophores. Cell-free apoptotic assays involving recombination of nuclear, cytosolic and mitochondrial fractions from the livers of wild type and Casp2−/− mice clearly point to a direct functional role for mitochondrial caspase-2 in apoptosis. Furthermore, cytochrome c release from Casp2−/− cells is decreased as compared with controls upon treatment with agents inducing mitochondrial dysfunction. Finally, we show that Casp2−/− primary skin fibroblasts are protected from oxidants that target the mitochondrial electron transport chain. Taken together, our results demonstrate that caspase-2 exists in the mitochondria and that it is essential for mitochondrial oxidative stress-induced apoptosis.

Original languageEnglish (US)
Article number16005
JournalCell Death Discovery
Volume2
Issue number1
DOIs
StatePublished - Dec 12 2016
Externally publishedYes

Bibliographical note

Funding Information:
We gratefully acknowledge Dr. Carol Troy, Columbia University College of Physicians and Surgeons, NY, USA, for providing us the Casp2−/− mice. We thank Dr. Victoria Centonze-Frohlich, St. Jude Children’s Research Hospital, for her contributions and guidance on the FRET experiments. This work was supported by the following grants from NIH (NIA): 5T32AG021890 (ML-C), 5R37AG007218 (BH) and 5R01AG007218 (JDL).

Publisher Copyright:
© 2016, The Author(s).

Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.

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