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.

The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore

Abstract

A sudden increase in permeability of the inner mitochondrial membrane, the so-called mitochondrial permeability transition, is a common feature of apoptosis and is mediated by the mitochondrial permeability transition pore (mtPTP). It is thought that the mtPTP is a protein complex formed by the voltage-dependent anion channel, members of the pro- and anti-apoptotic BAX-BCL2 protein family, cyclophilin D, and the adenine nucleotide (ADP/ATP) translocators (ANTs)1,2. The latter exchange mitochondrial ATP for cytosolic ADP and have been implicated in cell death. To investigate the role of the ANTs in the mtPTP, we genetically inactivated the two isoforms of ANT3,4,5 in mouse liver and analysed mtPTP activation in isolated mitochondria and the induction of cell death in hepatocytes. Mitochondria lacking ANT could still be induced to undergo permeability transition, resulting in release of cytochrome c. However, more Ca2+ than usual was required to activate the mtPTP, and the pore could no longer be regulated by ANT ligands. Moreover, hepatocytes without ANT remained competent to respond to various initiators of cell death. Therefore, ANTs are non-essential structural components of the mtPTP, although they do contribute to its regulation.

Your institute does not have access to this article

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: Preparation of a CRE-conditional Ant2-null mutant allele in mouse embryonic stem cells.
Figure 2: Inactivation of ANT in liver mitochondria.
Figure 3: Effect of ANT-deficiency on mtPTP activation in liver mitochondria.
Figure 4: Induction of cell death in ANT-deficient mouse hepatocytes.

References

  1. Zoratti, M. & Szabo, I. The mitochondrial permeability transition. Biochim. Biophys. Acta 1241, 139–176 (1995)

    Article  Google Scholar 

  2. Marzo, I. et al. Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. Science 281, 2027–2031 (1998)

    ADS  CAS  Article  Google Scholar 

  3. Levy, S. E., Chen, Y.-S., Graham, B. H. & Wallace, D. C. Expression and sequence analysis of the mouse adenine nucleotide translocase 1 and 2 genes. Gene 254, 57–66 (2000)

    CAS  Article  Google Scholar 

  4. Ellison, J. W., Salido, E. C. & Shapiro, L. J. Genetic mapping of the adenine nucleotide translocase-2 gene (Ant2) to the mouse proximal X chromosome. Genomics 36, 369–371 (1996)

    CAS  Article  Google Scholar 

  5. Graham, B. et al. A mouse model for mitochondrial myopathy and cardiomyopathy resulting from a deficiency in the heart/skeletal muscle isoform of the adenine nucleotide translocator. Nature Genet. 16, 226–234 (1997)

    CAS  Article  Google Scholar 

  6. Stepien, G., Torroni, A., Chung, A. B., Hodge, J. A. & Wallace, D. C. Differential expression of adenine nucleotide translocator isoforms in mammalian tissues and during muscle cell differentiation. J. Biol. Chem. 267, 14592–14597 (1992)

    CAS  PubMed  Google Scholar 

  7. Lunardi, J., Hurko, O., Engel, W. K. & Attardi, G. The multiple ADP/ATP translocase genes are differentially expressed during human muscle development. J. Biol. Chem. 267, 15267–15270 (1992)

    CAS  PubMed  Google Scholar 

  8. Postic, C. & Magnuson, M. A. DNA excision in liver by an albumin-Cre transgene occurs progressively with age. Genesis 26, 149–150 (2000)

    CAS  Article  Google Scholar 

  9. Boss, O., Hagen, T. & Lowell, B. B. Uncoupling proteins 2 and 3: potential regulators of mitochondrial energy metabolism. Diabetes 49, 143–156 (2000)

    CAS  Article  Google Scholar 

  10. Petronilli, V., Nicolli, A., Costantini, P., Colonna, R. & Bernardi, P. Regulation of the permeability transition pore, a voltage-dependent mitochondrial channel inhibited by cyclosporin A. Biochim. Biophys. Acta 1187, 255–259 (1994)

    CAS  Article  Google Scholar 

  11. Bernardi, P. Modulation of the mitochondrial cyclosporin A-sensitive permeability transition pore by the proton electrochemical gradient. Evidence that the pore can be opened by membrane depolarization. J. Biol. Chem. 267, 8834–8839 (1992)

    CAS  PubMed  Google Scholar 

  12. Esposito, L. A. et al. Mitochondrial oxidative stress in mice lacking the glutathione peroxidase-1 gene. Free Radic. Biol. Med. 28, 754–766 (2000)

    CAS  Article  Google Scholar 

  13. Kokoszka, J. E., Coskun, P., Esposito, L. & Wallace, D. C. Increased mitochondrial oxidative stress in the Sod2+/- mouse results in the age-related decline of mitochondrial function culminating in increased apoptosis. Proc. Natl Acad. Sci. USA 98, 2278–2283 (2001)

    ADS  CAS  Article  Google Scholar 

  14. Halestrap, A. P., Woodfield, K. Y. & Connern, C. P. Oxidative stress, thiol reagents, and membrane potential modulate the mitochondrial permeability transition by affecting nucleotide binding to the adenine nucleotide translocase. J. Biol. Chem. 272, 3346–3354 (1997)

    CAS  Article  Google Scholar 

  15. Lapidus, R. G. & Sokolove, P. M. The mitochondrial permeability transition. Interactions of spermine, ADP, and inorganic phosphate. J. Biol. Chem. 269, 18931–18936 (1994)

    CAS  PubMed  Google Scholar 

  16. Novgorodov, S. A., Gudz, T. I., Brierley, G. P. & Pfeiffer, D. R. Magnesium ion modulates the sensitivity of the mitochondrial permeability transition pore to cyclosporin A and ADP. Arch. Biochem. Biophys. 311, 219–228 (1994)

    CAS  Article  Google Scholar 

  17. Qian, T., Herman, B. & Lemasters, J. J. The mitochondrial permeability transition mediates both necrotic and apoptotic death of hepatocytes exposed to Br-A23187. Toxicol. Appl. Pharmacol. 154, 117–125 (1999)

    CAS  Article  Google Scholar 

  18. Hatano, E. et al. The mitochondrial permeability transition augments Fas-induced apoptosis in mouse hepatocytes. J. Biol. Chem. 275, 11814–11823 (2000)

    CAS  Article  Google Scholar 

  19. Trounce, I. A., Kim, Y. L., Jun, A. S. & Wallace, D. C. Assessment of mitochondrial oxidative phosphorylation in patient muscle biopsies, lymphoblasts, and transmitochondrial cell lines. Methods Enzymol. 264, 484–509 (1996)

    CAS  Article  Google Scholar 

  20. Leist, M. et al. Murine hepatocyte apoptosis induced in vitro and in vivo by TNF-α requires transcriptional arrest. J. Immunol. 153, 1778–1788 (1994)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank M. Magnuson for providing the Alb-Cre transgenic mice, L. Hayes for mouse husbandry and genotyping, and H. Yi for the electron microscope analysis. This work was funded by US National Institutes of Health grants awarded to D.C.W., G.R.M. and D.P.J.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Douglas C. Wallace.

Ethics declarations

Competing interests

J.E.K., K.G.W., S.E.L., J.E.S., G.R.M. and D.C.W. hold stock in a mitochondrial disease company, Medergy, Inc. but only D.C.W. holds significant equity as the scientific founder. The financial goals of the company do not conflict with the scientific content of this paper.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kokoszka, J., Waymire, K., Levy, S. et al. The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore. Nature 427, 461–465 (2004). https://doi.org/10.1038/nature02229

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature02229

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing