Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Paper
  • Published:

Mitochondrial cytochrome c release is caspase-dependent and does not involve mitochondrial permeability transition in didemnin B-induced apoptosis

Oncogene volume 20, pages 4085–4094 (2001)Cite this article

Abstract

Permeability transition, and a subsequent drop in mitochondrial membrane potential (ΔΨm), have been suggested to be mechanisms by which cytochrome c is released from the mitochondria into the cytosol during apoptosis. Furthermore, a drop in ΔΨm has been suggested to be an obligate early step in the apoptotic pathway. Didemnin B, a branched cyclic peptolide described to have immunosuppressive, anti-tumour, and anti-viral properties, induces rapid apoptosis in a range of mammalian cell lines. Induction of apoptosis by didemnin B in cultured human pro-myeloid HL-60 cells is the fastest and most complete ever described with all cells being apoptotic after 3 h of treatment. By utilizing the system of didemnin B-induced apoptosis in HL-60 cells, and the potent inhibitors of mitochondrial permeability transition, cyclosporin A and bongkrekic acid, we show that permeability transition as determined by changes in ΔΨm and mitochondrial Ca2+ fluxing, is not a requirement for apoptosis or cytochrome c release. In this system, changes in mitochondrial membrane potential and cytochrome c release are shown to be dependent on caspase activation, and to occur concurrently with the release of caspase-9 from mitochondria, genomic DNA fragmentation and apoptotic body formation.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10

Similar content being viewed by others

Abbreviations

AIF:

apoptosis inducing factor

CyA:

cyclosporin A

BA:

bongkrekic acid

JC-1:

5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolcarbocyanine iodide

PT:

permeability transition

PTP:

permeability transition pore

CCCP:

carbonyl cyanide m-chlorophenyl hydrazone

DAPI:

4,6′-diamidino-2-phenylindole

mtDNA:

mitochondrial DNA

z-VAD-fmk:

benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone

Ac-DEVD-cho:

acetyl-Asp-Glu-Val-L-Asp-aldehyde

Ac-YVAD-cmk:

acetyl-Tyr-Val-Ala-Asp-chloromethylketone

References

  • Bradford MM . 1976 Anal. Biochem. 72: 248–254

  • Brenner C, Cadiou H, Vieira HL, Zamzami N, Marzo I, Xie Z, Leber B, Andrews D, Duclohier H, Reed JC, Kroemer G . 2000 Oncogene 19: 329–336

  • Bossy-Wetzel E, Newmeyer DD, Green DR . 1998 EMBO J. 17: 37–49

  • Chen Q, Gong B, Almasan A . 2000 Cell. Death Differ. 7: 227–233

  • Crews CM, Collins JL, Lane WS, Snapper ML, Schreiber SL . 1994 J. Biol. Chem. 269: 15411–15414

  • Crews CM, Lane WS, Schreiber SL . 1996 Proc. Natl. Acad. Sci. USA 93: 4316–4319

  • de Bruijn J, Frost DJ, Nugteren DH, Graudemer A, Lijmbach GWM, Cox HC, Berends W . 1973 Tetrahedron 29: 1541–1547

  • Desagher S, Martinou JC . 2000 Trends Cell. Biol. 10: 369–377

  • Fournier N, Ducet G, Crevat A . 1987 J. Bioenerg. Biomembr. 19: 297–303

  • Gamen S, Anel A, Montoya J, Marzo I, Piñeiro A, Naval J . 1995 FEBS Lett. 376: 15–18

  • Gamen S, Anel A, Perez-Galan P, Lasierra P, Johnson D, Pineiro A, Naval J . 2000 Exp. Cell. Res. 258: 223–235

  • Goldstein JC, Waterhouse NJ, Juin P, Evan GI, Green DR . 2000 Nat. Cell. Biol. 2: 156–162

  • Grubb DR, Wolvetang EJ, Lawen A . 1995 Biochem. Biophys. Res. Commun. 215: 1130–1136

  • Henderson PJ, Lardy HA . 1970 J. Biol. Chem. 245: 1319–1326

  • Herrmann M, Lorenz H-M, Voll R, Grünke M, Woith W, Kalden JR . 1994 Nucl. Acid. Res. 22: 5506–5507

  • Ichas F, Mazat JP . 1998 Biochim. Biophys. Acta 1366: 33–50

  • Jacobson MD, Burne JF, King MP, Miyashita T, Reed JC, Raff MC . 1993 Nature 361: 365–369

  • Jiang S, Cai J, Wallace DC, Jones DP . 1999 J. Biol. Chem. 274: 29905–29911

  • Johnson KL, Vaillant F, Lawen A . 1996 FEBS Lett. 383: 1–5

  • Johnson KL, Grubb DR, Lawen A . 1999 J. Cell. Biochem. 72: 269–278

  • Johnson KL, Lawen A . 1999 Immunol. Cell. Biol. 77: 242–248

  • Kim TH, Zhao Y, Barber MJ, Kuharsky DK, Yin XM . 2000 J. Biol. Chem. 275: 39474–39481

  • Kirsch DG, Doseff A, Chau BN, Lim DS, de Souza-Pinto NC, Hansford R, Kastan MB, Lazebnik YA, Hardwick JM . 1999 J. Biol. Chem. 274: 21155–21161

  • Klingenberg M, Grebe K, Heldt HW . 1970 Biochem. Biophys. Res. Commun. 39: 344–351

  • Kluck RM, Bossy-Wetzel E, Green DR, Newmeyer DD . 1997a Science 275: 1132–1136

  • Kluck RM, Martin SJ, Hoffman BM, Zhou JS, Green DR, Newmeyer DD . 1997b EMBO J. 16: 4639–4649

  • Krajewski S, Krajewski M, Ellerby LM, Welsh K, Xie Z, Deveraux QL, Salvesen GS, Bredesen DE, Rosenthal RE, Fiskum G, Reed JC . 1999 Proc. Natl. Acad. Sci. USA 96: 5752–5757

  • Krohn AJ, Wahlbrink T, Prehn JH . 1999 J. Neurosci. 19: 7394–7404

  • Larm JA, Vaillant F, Linnane AW, Lawen A . 1994 J. Biol. Chem. 269: 30097–30100

  • Li P, Nijhawan D, Budihardjo I, Srinivasula SM, Ahmad M, Alnemri ES, Wang X . 1997 Cell 91: 479–489

  • Lijmbach GWM, Cox HC, Berends W . 1970 Tetrahedron 26: 5993–5999

  • Liu X, Kim CN, Yang J, Jemmerson R, Wang X . 1996 Cell 86: 147–157

  • Marchetti P, Castedo M, Susin SA, Zamzami N, Hirsch T, Macho A, Haeffner A, Hirsch F, Geuskens M, Kroemer G . 1996 J. Exp. Med. 184: 1155–1160

  • Meng L, Sin N, Crews CM . 1998 Biochemistry 37: 10488–10492

  • Montgomery DW, Zukoski CF . 1985 Transplantation 40: 49–56

  • Pastorino JG, Chen S-T, Tafani M, Snyder JW, Farber JL . 1998 J. Biol. Chem. 273: 7770–7775

  • Petronilli V, Cola C, Massari S, Colonna R, Bernardi P . 1993 J. Biol. Chem. 268: 21939–21945

  • Rinehart Jr KL, Gloer JB, Wilson GR, Hughes Jr RG, Li LH, Renis HE, McGoven JP . 1983 Fed. Proc. 42: 87–90

  • Robertson JD, Gogvadze V, Zhivotovsky B, Orrenius S . 2000 J. Biol. Chem. 275: 32438–32443

  • Russel WC, Newman C, Williamson DH . 1975 Nature 253: 4671–462

  • Rytomaa M, Lehmann K, Downward J . 2000 Oncogene 19: 4461–4468

  • Saito M, Korsmeyer SJ, Schlesinger PH . 2000 Nat. Cell. Biol. 2: 553–555

  • Salvioli S, Ardizzoni A, Franceschi C, Cossarizza A . 1997 FEBS Lett. 411: 77–82

  • Scarlett JL, Murphy MP . 1997 FEBS Lett. 418: 282–286

  • Scorrano L, Petronilli V, Di Lisa F, Bernardi P . 1999 J. Biol. Chem. 274: 22581–22585

  • Slee EA, Harte MT, Kluck RM, Wolf BB, Casiano CA, Newmeyer DD, Wang HG, Reed JC, Nicholson DW, Alnemri ES, Green DR, Martin SJ . 1999 J Cell Biol. 144: 281–292

  • Slee EA, Keogh SA, Martin SJ . 2000 Cell Death Differ. 7: 556–565

  • Susin SA, Lorenzo HK, Zamzami N, Marzo I, Snow BE, Brothers GM, Mangion J, Jacotot E, Costantini P, Loeffler M, Larochette N, Goodlett DR, Aebersold R, Siderovski DP, Penninger JM, Kroemer G . 1999 Nature 397: 441–446

  • Vaillant F, Nagley P . 1995 Hum. Mol. Genet. 4,: 903–914

  • Weed SD, Stringfellow DA . 1983 Antiviral Res. 3: 269–274

  • Yang J, Liu X, Bhalla K, Kim CN, Ibrado AM, Cai J, Peng T-I, Jones DP, Wang X . 1997 Science 275: 1129–1132

  • Zamzami N, Marchetti P, Castedo M, Decaudin D, Macho A, Hirsch T, Susin SA, Petit PX, Mignotte B, Kroemer G . 1995a J. Exp. Med. 182: 367–377

  • Zamzami N, Marchetti P, Castedo M, Hirsch T, Susin SA, Masse B, Kroemer G . 1996 FEBS Lett. 384: 53–57

  • Zamzami N, Marchetti P, Castedo M, Zanin C, Vayssiere JL, Petit PX, Kroemer G . 1995b J. Exp. Med. 181: 1661–1672

  • Zhai D, Huang X, Han X, Yang F . 2000 FEBS Lett. 472: 293–296

  • Zhuang J, Dinsdale D, Cohen GM . 1998 Cell Death Differ. 5: 953–962

  • Zou H, Henzel WJ, Liu X, Lutschg A, Wang X . 1997 Cell 90: 405–413

Download references

Acknowledgements

We thank the National Cancer Institute, Drug Synthesis and Chemistry Branch, Bethesda, (USA) for the gift of didemnin B, Prof JA Duine from Delft University of Technology (Netherlands) for the gift of bongkrekic acid, Dr René Traber from Novartis Pharma Ltd (Basel, Switzerland) for the gift of cyclosporin A, Dr Donald Nicholson from Department of Biochemistry and Molecular Biology, Merck Frosst Centre for Therapeutic Research, Quebec, Canada for the gift of anti-caspase-3 antibody. We also thank Mr Barry Veitch for his invaluable assistance with the electron microscopy work.

Author information

Author notes

  1. François Vaillant

    Present address: Department of Veterinary Pathology, University of Glasgow Veterinary School, Bearsden Road, Glasgow, G61 1QH, UK

  2. Jennifer D Ly and François Vaillant: JD Ly and F Vaillant contributed equally to this publication.

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, School of Biomedical Sciences, Monash University, Melbourne, 3800, Australia

    David R Grubb, Jennifer D Ly, Karina L Johnson & Alfons Lawen

Authors
  1. David R Grubb
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jennifer D Ly
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. François Vaillant
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Karina L Johnson
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alfons Lawen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Alfons Lawen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Grubb, D., Ly, J., Vaillant, F. et al. Mitochondrial cytochrome c release is caspase-dependent and does not involve mitochondrial permeability transition in didemnin B-induced apoptosis. Oncogene 20, 4085–4094 (2001). https://doi.org/10.1038/sj.onc.1204545

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1204545

Keywords

This article is cited by

Search

Advanced search

Quick links