Abstract
Most cell death stimuli trigger the mitochondrial release of cytochrome c and other cofactors that induce caspase activation and ensuing apoptosis. Apoptosis is also associated with massive mitochondrial fragmentation and cristae remodeling. Dynamin-related protein 1 (Drp1), a protein of the mitochondrial fission machinery, has been reported to participate in apoptotic mitochondrial fragmentation. Several theories explaining the mechanisms of cytochome c release have been proposed. One suggests that it relies on the activation of Drp1-mediated mitochondrial fission. Here, we report that downregulation of Drp1 inhibits fragmentation of the mitochondrial network and partially prevents the release of cytochrome c but fails to prevent the release of other mitochondrial factors such as second mitochondria-derived activator of caspase/direct IAP-binding protein with low pI, Omi/HtrA2, adenylate kinase 2 and deafness dystonia peptide/TIMM8a. An explanation for the prevention of cytochrome c release is provided by our observation that inhibiting Drp1-mediated mitochondrial fission prevents the mitochondrial release of soluble OPA1 that was proposed to regulate cristae remodeling and complete cytochrome c release during apoptosis. Finally, we observed that downregulation of Drp1 delays but does not inhibit apoptosis, suggesting that mitochondrial fragmentation is not a prerequisite for apoptosis.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- AK2:
-
adenylate kinase 2
- DDP:
-
deafness dystonia peptide
- DIABLO:
-
direct IAP-binding protein with low pI
- Drp1:
-
dynamin-related protein 1
- IMM:
-
inner-mitochondrial membrane
- IMS:
-
intermembrane space
- Mfn:
-
mitofusin
- MOMP:
-
mitochondrial outer membrane permeabilization
- PARP:
-
poly(ADP-ribose) polymerase
- PCD:
-
programmed cell death
- Smac:
-
second mitochondria-derived activator of caspase
References
Riedl SJ, Shi Y . Molecular mechanisms of caspase regulation during apoptosis. Nat Rev Mol Cell Biol 2004; 5: 897–907.
Wang X . The expanding role of mitochondria in apoptosis. Genes Dev 2001; 15: 2922–2933.
Green DR, Kroemer G . The pathophysiology of mitochondrial cell death. Science 2004; 305: 626–629.
Frank S, Gaume B, Bergmann-Leitner ES, Leitner WW, Robert EG, Catez F et al. The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev Cell 2001; 1: 515–525.
Youle RJ, Karbowski M . Mitochondrial fission in apoptosis. Nat Rev Mol Cell Biol 2005; 6: 657–663.
Breckenridge DG, Stojanovic M, Marcellus RC, Shore GC . Caspase cleavage product of BAP31 induces mitochondrial fission through endoplasmic reticulum calcium signals, enhancing cytochrome c release to the cytosol. J Cell Biol 2003; 160: 1115–1127.
Karbowski M, Lee YJ, Gaume B, Jeong SY, Frank S, Nechushtan A et al. Spatial and temporal association of Bax with mitochondrial fission sites, Drp1, and Mfn2 during apoptosis. J Cell Biol 2002; 159: 931–938.
Okamoto K, Shaw JM . Mitochondrial morphology and dynamics in yeast and multicellular eukaryotes. Annu Rev Genet 2005; 39: 503–536.
James DI, Parone PA, Mattenberger Y, Martinou JC . hFis1, a novel component of the mammalian mitochondrial fission machinery. J Biol Chem 2003; 278: 36373–36379.
Smirnova E, Shurland DL, Ryazantsev SN, van der Bliek AM . A human dynamin-related protein controls the distribution of mitochondria. J Cell Biol 1998; 143: 351–358.
Smirnova E, Griparic L, Shurland DL, van der Bliek AM . Dynamin-related protein Drp1 is required for mitochondrial division in mammalian cells. Mol Biol Cell 2001; 12: 2245–2256.
Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC . Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J Cell Biol 2003; 160: 189–200.
Olichon A, Baricault L, Gas N, Guillou E, Valette A, Belenguer P et al. Loss of OPA1 perturbates the mitochondrial inner membrane structure and integrity, leading to cytochrome c release and apoptosis. J Biol Chem 2003; 278: 7743–7746.
Arnoult D, Grodet A, Lee YJ, Estaquier J, Blackstone C . Release of OPA1 during apoptosis participates in the rapid and complete release of cytochrome c and subsequent mitochondrial fragmentation. J Biol Chem 2005; 280: 35742–35750.
Delettre C, Griffoin JM, Kaplan J, Dollfus H, Lorenz B, Faivre L et al. Mutation spectrum and splicing variants in the OPA1 gene. Hum Genet 2001; 109: 584–591.
Frezza C, Cipolat S, Martins de Brito O, Micaroni M, Beznoussenko GV, Rudka T et al. OPA1 controls apoptotic cristae remodeling independently from mitochondrial fusion. Cell 2006; 126: 177–189.
Esseiva AC, Chanez AL, Bochud-Allemann N, Martinou JC, Hemphill A, Schneider A . Temporal dissection of Bax-induced events leading to fission of the single mitochondrion in Trypanosoma brucei. EMBO Rep 2004; 5: 268–273.
Gao W, Pu Y, Luo KQ, Chang DC . Temporal relationship between cytochrome c release and mitochondrial swelling during UV-induced apoptosis in living HeLa cells. J Cell Sci 2001; 114: 2855–2862.
Arnoult D . Mitochondrial fragmentation in apoptosis. Trends Cell Biol 2007; 17: 6–12.
Martinou JC, Youle RJ . Which came first, the cytochrome c release or the mitochondrial fission? Cell Death Differ 2006; 13: 1291–1295.
Parone PA, Martinou JC . Mitochondrial fission and apoptosis: an ongoing trial. Biochim Biophys Acta 2006; 1763: 522–530.
Bauer MF, Hofmann S, Neupert W, Brunner M . Protein translocation into mitochondria: the role of TIM complexes. Trends Cell Biol 2000; 10: 25–31.
Arnoult D, Rismanchi N, Grodet A, Roberts RG, Seeburg DP, Estaquier J et al. Bax/Bak-dependent release of DDP/TIMM8a promotes Drp1-mediated mitochondrial fission and mitoptosis during programmed cell death. Curr Biol 2005; 15: 2112–2118.
Single B, Leist M, Nicotera P . Simultaneous release of adenylate kinase and cytochrome c in cell death. Cell Death Differ 1998; 5: 1001–1003.
Germain M, Mathai JP, McBride HM, Shore GC . Endoplasmic reticulum BIK initiates DRP1-regulated remodelling of mitochondrial cristae during apoptosis. EMBO J 2005; 24: 1546–1556.
Lee YJ, Jeong SY, Karbowski M, Smith CL, Youle RJ . Roles of the mammalian mitochondrial fission and fusion mediators Fis1, Drp1, and Opa1 in apoptosis. Mol Biol Cell 2004; 15: 5001–5011.
Sugioka R, Shimizu S, Tsujimoto Y . Fzo1, a protein involved in mitochondrial fusion, inhibits apoptosis. J Biol Chem 2004; 279: 52726–52734.
Parone PA, James DI, Da Cruz S, Mattenberger Y, Donze O, Barja F et al. Inhibiting the mitochondrial fission machinery does not prevent Bax/Bak-dependent apoptosis. Mol Cell Biol 2006; 26: 7397–7408.
Wei MC, Zong WX, Cheng EH, Lindsten T, Panoutsakopoulou V, Ross AJ et al. Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science 2001; 292: 727–730.
Chen H, Chomyn A, Chan DC . Disruption of fusion results in mitochondrial heterogeneity and dysfunction. J Biol Chem 2005; 280: 26185–26192.
Scorrano L, Ashiya M, Buttle K, Weiler S, Oakes SA, Mannella CA et al. A distinct pathway remodels mitochondrial cristae and mobilizes cytochrome c during apoptosis. Dev Cell 2002; 2: 55–67.
Cipolat S, Rudka T, Hartmann D, Costa V, Serneels L, Craessaerts K et al. Mitochondrial rhomboid PARL regulates cytochrome c release during apoptosis via OPA1-dependent cristae remodeling. Cell 2006; 126: 163–175.
Ishihara N, Fujita Y, Oka T, Mihara K . Regulation of mitochondrial morphology through proteolytic cleavage of OPA1. EMBO J 2006; 25: 2966–2977.
Uren RT, Dewson G, Bonzon C, Lithgow T, Newmeyer DD, Kluck RM . Mitochondrial release of pro-apoptotic proteins: electrostatic interactions can hold cytochrome c but not Smac/DIABLO to mitochondrial membranes. J Biol Chem 2005; 280: 2266–2274.
Ott M, Robertson JD, Gogvadze V, Zhivotovsky B, Orrenius S . Cytochrome c release from mitochondria proceeds by a two-step process. Proc Natl Acad Sci USA 2002; 99: 1259–1263.
Petrosillo G, Ruggiero FM, Pistolese M, Paradies G . Reactive oxygen species generated from the mitochondrial electron transport chain induce cytochrome c dissociation from beef-heart submitochondrial particles via cardiolipin peroxidation. Possible role in the apoptosis. FEBS Lett 2001; 509: 435–438.
Delivani P, Adrain C, Taylor RC, Duriez PJ, Martin SJ . Role for CED-9 and Egl-1 as regulators of mitochondrial fission and fusion dynamics. Mol Cell 2006; 21: 761–773.
Brustugun OT, Fladmark KE, Doskeland SO, Orrenius S, Zhivotovsky B . Apoptosis induced by microinjection of cytochrome c is caspase-dependent and is inhibited by Bcl-2. Cell Death Differ 1998; 5: 660–668.
Arnoult D, Gaume B, Karbowski M, Sharpe JC, Cecconi F, Youle RJ . Mitochondrial release of AIF and EndoG requires caspase activation downstream of Bax/Bak-mediated permeabilization. EMBO J 2003; 22: 4385–4399.
Acknowledgements
We are very grateful to Dr. Anne Lombès and for her valuable assistance in oxygen consumption studies and to Dr. Craig blackstone for critical reviews of our manuscript, respectively. We thank Dr. Richard Youle for the gift or control RNAi and Drp1 RNAi constructs and for discussions. Special thanks to Dr. Philippe Parone and Dr. Jean Claude Martinou for stimulating scientific discussions and for sharing unpublished data. We apologize to the authors of relevant articles that are not cited because of space limitations. This work was funded by grants from the Agence Nationale de Recherche sur le Sida (ANRS).
Author information
Authors and Affiliations
Corresponding author
Additional information
Edited by DR Green
Supplementary Information accompanies the paper on Cell Death and Differentiation website (http://www.nature.com/cdd)
Supplementary information
Rights and permissions
About this article
Cite this article
Estaquier, J., Arnoult, D. Inhibiting Drp1-mediated mitochondrial fission selectively prevents the release of cytochrome c during apoptosis. Cell Death Differ 14, 1086–1094 (2007). https://doi.org/10.1038/sj.cdd.4402107
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1038/sj.cdd.4402107
