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GAPDH, a novel regulator of the pro-apoptotic mitochondrial membrane permeabilization

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Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a pleiotropic enzyme that is overexpressed in apoptosis and in several human chronic pathologies. Here, we report that the protein accumulates in mitochondria during apoptosis, and induces the pro-apoptotic mitochondrial membrane permeabilization, a decisive event of the intrinsic pathway of apoptosis. GAPDH was localized by immunogold labeling and identified by matrix-assisted laser desorption/ionization-time of flight and nano liquid chromatography mass spectroscopy/mass spectroscopy in the mitochondrion of various tissues and origins. In isolated mitochondria, GAPDH can be imported and interact with the voltage-dependent anion channel (VDAC1), but not the adenine nucleotide translocase (ANT). The protein mediates a cyclosporin A-inhibitable permeability transition, characterized by a loss of the inner transmembrane potential, matrix swelling, permeabilization of the inner mitochondrial membrane and the release of two pro-apoptotic proteins, cytochrome c and apoptosis-inducing factor (AIF). This novel function of GAPDH might have implications for the understanding of mitochondrial biology, oncogenesis and apoptosis.

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Abbreviations

AIF:

apoptosis-inducing factor

ANT:

adenine nucleotide translocator

Cyt c:

cytochrome c

CsA:

cyclosporin A

DIDS:

4,4′-diisothiocyanatostilbene-2,2′-disulfonate

DTT:

dithiotheritol

m:

mitochondrial transmembrane potential

GAPDH:

glyceraldehyde-3-phosphate dehydrogenase

mClCCP:

carbonylcyanide m-chlorophenylhydrazone

MMP:

mitochondrial membrane permeabilization

MUP:

4-methylumbelliferyl phosphate

nanoLC MS/MS:

nano liquid chromatography mass spectroscopy/mass spectroscopy

IM:

inner membrane

OM:

outer membrane

PI:

propidium iodide

ROS:

reactive oxygen species

PTPC:

permeability transition pore complex

PBR:

peripheral benzodiazepine receptor

VDAC:

voltage-dependent anion channel

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Acknowledgements

We thank C Henry (platform PAPSS, INRA, http://www.jouy.inra.fr/unites/proteines/papss/) and C Longin (platform MIMA2, http://voxel.jouy.inra.fr/mima2) for their professional assistance in mass spectrometry and electron microsocopy, respectively. This work was supported by grants funded by ARC, CNRS and French Ministry of Research. (to CB), by a special grant of the League against Cancer and by the European Union (Trans-Death, RIGHT) (to GK). MLB received a postdoctoral fellowship of CNRS. AT and AD received a doctoral and post doctoral fellowships from the Ligue Contre le Cancer.

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Authors and Affiliations

  1. CNRS UMR 8159, Université de Versailles/SQY, Versailles, France

    A Tarze, A Deniaud, M Le Bras, E Maillier & C Brenner

  2. INRA UMR STLO, Rennes, France

    D Molle & G Jan

  3. IGR, CNRS UMR 8125, Villejuif, France

    N Larochette, N Zamzami & G Kroemer

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  1. A Tarze
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  2. A Deniaud
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  3. M Le Bras
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  4. E Maillier
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  5. D Molle
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  6. N Larochette
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  7. N Zamzami
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  8. G Jan
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  9. G Kroemer
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  10. C Brenner
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Correspondence to C Brenner.

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Tarze, A., Deniaud, A., Le Bras, M. et al. GAPDH, a novel regulator of the pro-apoptotic mitochondrial membrane permeabilization. Oncogene 26, 2606–2620 (2007). https://doi.org/10.1038/sj.onc.1210074

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