Abstract
Non-muscle cofilin (n-cofilin) is a member of the ADF/cofilin family of actin depolymerizing proteins. Recent studies reported a mitochondrial translocation of n-cofilin during apoptosis. As these studies also revealed impaired cytochrome c release and a block in apoptosis upon small interfering RNA-mediated n-cofilin knockdown, n-cofilin was postulated to be essential for apoptosis induction. To elucidate the general importance of ADF/cofilin activity for apoptosis, we exposed mouse embryonic fibroblasts deficient for n-cofilin, ADF (actin depolymerizing factor), or all ADF/cofilin isoforms to well-characterized apoptosis inducers. Cytochrome c release, caspase-3 activation, and apoptotic chromatin condensation were unchanged in all mutant fibroblasts. Thus, we conclude that ADF/cofilin activity is not generally required for induction or progression of apoptosis in mammalian cells. Interestingly, mitochondrial association of ADF and n-cofilin during apoptosis was preceded by, and dependent on, actin that translocated by a yet unknown mechanism to mitochondria during cell death.
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Abbreviations
- 3T3:
-
3-day transfer protocol
- AC-MEF:
-
MEF-deficient for ADF and n-cofilin
- ADF:
-
actin-depolymerizing factor
- COX II:
-
cytochrome c oxidase subunit II
- E:
-
embryonic
- flx:
-
floxed
- JAS:
-
jasplakinolide
- MCM:
-
OH-TAM-inducible cre recombinase
- m-cofilin:
-
muscle cofilin
- MEF:
-
mouse embryonic fibroblast
- MSH buffer:
-
mannitol–sucrose–hepes–buffer
- n-Cof−/− MEF:
-
MEFs deficient for n-cofilin
- n-Cofflx/flx MEF:
-
MEFs with floxed n-cofilin gene
- n-cofilin:
-
non-muscle cofilin
- OH-TAM:
-
4-hydroxytamoxifen
- P:
-
passage
- PGAM:
-
phosphoglycerate mutase
- PHAL:
-
phalloidin
- phospho-AC:
-
phosphorylated ADF/cofilin
- siRNA:
-
small interfering RNA
- SMAC:
-
second mitochondria-derived activator of caspase
- STS:
-
staurosporine
- WT:
-
wild type
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Acknowledgements
We thank Dr. W Witke (Bonn/Germany) for providing ADF and n-Cofflx/flx mice, Dr. JM Herrmann (Kaiserslautern/Germany) for support, Dr. M Reth (Freiburg/Germany) for providing the MCM cassette, and Drs S Orrenius and B Zhivotovsky (Stockholm/Sweden) for critical reading the manuscript. This work was funded by the Research Initiative Membrane Transport (RIMB) of the University of Kaiserslautern. KR is a recipient of a doctoral fellowship from the Stipendienstiftung des Landes Rheinland-Pfalz.
Author Contributions
Experiments were designed and results were discussed by KR, MO, and MBR. Mutant mice were generated by CBG. MEFs were generated by KR and MC. Data analysis was performed by KR. Manuscript was written by MO, EF, and MBR.
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Rehklau, K., Gurniak, C., Conrad, M. et al. ADF/cofilin proteins translocate to mitochondria during apoptosis but are not generally required for cell death signaling. Cell Death Differ 19, 958–967 (2012). https://doi.org/10.1038/cdd.2011.180
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DOI: https://doi.org/10.1038/cdd.2011.180
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