Abstract
Cardiolipin (CL) is a mitochondria-specific phospholipid synthesized by CL synthase (CLS). We describe here a human gene for CLS and its analysis via RNAi knockdown on apoptotic progression. Although mitochondrial membrane potential is unchanged in cells containing only 25% of the normal amount of CL, free cytochrome c (cyt. c) is detected in the intermembrane space and the mitochondria exhibit signs of reorganized cristae. However, the release of cyt. c from the mitochondria still requires apoptotic stimulation. Increased sensitivity to apoptotic signals and accelerated rates of apoptosis are observed in CL-deficient cells, followed by elevated levels of secondary necrosis. Apoptosis is thought to progress via binding of truncated Bid (tBid) to mitochondrial CL, followed by CL oxidation which results in cyt. c release. The exaggerated and accelerated apoptosis observed in CL-deficient cells is matched by an accelerated reduction in membrane potential and increased cyt. c release, but not by decreased tBid binding. This study suggests that the CL/cyt. c relationship is important in apoptotic progression and that regulating CL oxidation or/and deacylation could represent a possible therapeutic target.
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Abbreviations
- CDP:
-
cytidine 5′-diphosphate
- Chx:
-
cycloheximide
- CL:
-
cardiolipin
- CLS:
-
cardiolipin synthase
- CMP:
-
cytidine 5′-monophosphate
- cyt. c:
-
cytochrome c
- DHB:
-
2,5-dihydroxybenzoic acid
- MLS:
-
mitochondrial localization sequence
- PA:
-
phosphatidic acid
- PG:
-
phosphatidylglycerol
- PGP:
-
phosphatidylglycerophosphate
- STS:
-
staurosporine
- tBid:
-
truncated Bid
- TLC:
-
thin layer chromatography
- TNF-α:
-
tumor necrosis factor-α
References
Hatch GM . Cell biology of cardiac mitochondrial phospholipids. Biochem Cell Biol 2004; 82: 99–112.
Schlame M, Rua D, Greenberg ML . The biosynthesis and functional role of cardiolipin. Prog Lipid Res 2000; 39: 257–288.
Chang SC, Heacock PN, Mileykovskaya E, Voelker DR, Dowhan W . Isolation and characterization of the gene (CLS1) encoding cardiolipin synthase in Saccharomyces cerevisiae. J Biol Chem 1998; 273: 14933–14941.
Zhong Q, Gohil VM, Ma L, Greenberg ML . Absence of cardiolipin results in temperature sensitivity, respiratory defects, and mitochondrial DNA instability independent of pet56. J Biol Chem 2004; 279: 32294–32300.
Tuominen EK, Wallace CJ, Kinnunen PK . Phospholipid–cytochrome c interaction: evidence for the extended lipid anchorage. J Biol Chem 2002; 277: 8822–8826.
Spooner PJ, Watts A . Cytochrome c interactions with cardiolipin in bilayers: a multinuclear magic-angle spinning NMR study. Biochemistry 1992; 31: 10129–10138.
Zhang M, Mileykovskaya E, Dowhan W . Gluing the respiratory chain together. Cardiolipin is required for supercomplex formation in the inner mitochondrial membrane. J Biol Chem 2002; 277: 43553–43556.
Robinson NC . Functional binding of cardiolipin to cytochrome c oxidase. J Bioenerg Biomembr 1993; 25: 153–163.
Wright MM, Howe AG, Zaremberg V . Cell membranes and apoptosis: role of cardiolipin, phosphatidylcholine, and anticancer lipid analogues. Biochem Cell Biol 2004; 82: 18–26.
Gonzalvez F, Bessoule JJ, Rocchiccioli F, Manon S, Petit PX . Role of cardiolipin on tBid and tBid/Bax synergistic effects on yeast mitochondria. Cell Death Differ 2005; 12: 659–667.
Gonzalvez F, Pariselli F, Dupaigne P, Budihardjo I, Lutter M, Antonsson B et al. tBid interaction with cardiolipin primarily orchestrates mitochondrial dysfunctions and subsequently activates Bax and Bak. Cell Death Differ 2005; 12: 614–626.
Lutter M, Fang M, Luo X, Nishijima M, Xie X, Wang X . Cardiolipin provides specificity for targeting of tBid to mitochondria. Nat Cell Biol 2000; 2: 754–761.
Lutter M, Perkins GA, Wang X . The pro-apoptotic Bcl-2 family member tBid localizes to mitochondrial contact sites. BMC Cell Biol 2001; 2: 22.
Orrenius S, Zhivotovsky B . Cardiolipin oxidation sets cytochrome c free. Nat Chem Biol 2005; 1: 188–189.
Gross A, Yin XM, Wang K, Wei MC, Jockel J, Milliman C et al. Caspase cleaved BID targets mitochondria and is required for cytochrome c release, while BCL-XL prevents this release but not tumor necrosis factor-R1/Fas death. J Biol Chem 1999; 274: 1156–1163.
Epand RF, Martinou JC, Fornallaz-Mulhauser M, Hughes DW, Epand RM . The apoptotic protein tBid promotes leakage by altering membrane curvature. J Biol Chem 2002; 277: 32632–32639.
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.
Williams JG, McMaster CR . Scanning alanine mutagenesis of the CDP-alcohol phosphotransferase motif of Saccharomyces cerevisiae cholinephosphotransferase. J Biol Chem 1998; 273: 13482–13487.
Barth PG, Valianpour F, Bowen VM, Lam J, Duran M, Vaz FM et al. X-linked cardioskeletal myopathy and neutropenia (Barth syndrome): an update. Am J Med Genet A 2004; 126: 349–354.
Ostrander DB, Sparagna GC, Amoscato AA, McMillin JB, Dowhan W . Decreased cardiolipin synthesis corresponds with cytochrome c release in palmitate-induced cardiomyocyte apoptosis. J Biol Chem 2001; 276: 38061–38067.
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.
Iverson SL, Orrenius S . The cardiolipin–cytochrome c interaction and the mitochondrial regulation of apoptosis. Arch Biochem Biophys 2004; 423: 37–46.
Nakagawa Y . Initiation of apoptotic signal by the peroxidation of cardiolipin of mitochondria. Ann N Y Acad Sci 2004; 1011: 177–184.
Wronski R, Golob N, Grygar E, Windisch M . Two-color, fluorescence-based microplate assay for apoptosis detection. Biotechniques 2002; 32: 666–668.
Hu X, Han Z, Wyche JH, Hendrickson EA . Helix 6 of tBid is necessary but not sufficient for mitochondrial binding activity. Apoptosis 2003; 8: 277–289.
Kawasaki K, Kuge O, Chang SC, Heacock PN, Rho M, Suzuki K et al. Isolation of a Chinese hamster ovary (CHO) cDNA encoding phosphatidylglycerophosphate (PGP) synthase, expression of which corrects the mitochondrial abnormalities of a PGP synthase-defective mutant of CHO-K1 cells. J Biol Chem 1999; 274: 1828–1834.
Ohtsuka T, Nishijima M, Suzuki K, Akamatsu Y . Mitochondrial dysfunction of a cultured Chinese hamster ovary cell mutant deficient in cardiolipin. J Biol Chem 1993; 268: 22914–22919.
Kagan VE, Tyurin VA, Jiang J, Tyurina YY, Ritov VB, Amoscato AA et al. Cytochrome c acts as a cardiolipin oxygenase required for release of proapoptotic factors. Nat Chem Biol 2005; 1: 223–232.
Shidoji Y, Hayashi K, Komura S, Ohishi N, Yagi K . Loss of molecular interaction between cytochrome c and cardiolipin due to lipid peroxidation. Biochem Biophys Res Commun 1999; 264: 343–347.
Zhang M, Mileykovskaya E, Dowhan W . Cardiolipin is essential for organization of complexes III and IV into a supercomplex in intact yeast mitochondria. J Biol Chem 2005; 280: 29403–29408.
Valianpour F, Mitsakos V, Schlemmer D, Towbin JA, Taylor JM, Ekert PG et al. Monolysocardiolipins accumulate in Barth syndrome but do not lead to enhanced apoptosis. J Lipid Res 2005; 46: 1182–1195.
Ludovico P, Madeo F, Silva M . Yeast programmed cell death: an intricate puzzle. IUBMB Life 2005; 57: 129–135.
Brummelkamp TR, Bernards R, Agami R . A system for stable expression of short interfering RNAs in mammalian cells. Science 2002; 296: 550–553.
Valianpour F, Wanders RJ, Barth PG, Overmars H, van Gennip AH . Quantitative and compositional study of cardiolipin in platelets by electrospray ionization mass spectrometry: application for the identification of Barth syndrome patients. Clin Chem 2002; 48: 1390–1397.
Chretien D, Benit P, Chol M, Lebon S, Rotig A, Munnich A et al. Assay of mitochondrial respiratory chain complex I in human lymphocytes and cultured skin fibroblasts. Biochem Biophys Res Commun 2003; 301: 222–224.
Darin N, Kadhom N, Briere JJ, Chretien D, Bebear CM, Rotig A et al. Mitochondrial activities in human cultured skin fibroblasts contaminated by Mycoplasma hyorhinis. BMC Biochem 2003; 4: 15.
Acknowledgements
We thank D. Bogenhagen, W. Dowhan, E.Y. Kim, J. Martinou, U. Moll, A. Neiman, R. Sternglanz, A. Sutton, S. Tsirka, R. Houtkooper, and members of the Frohman lab for technical advice and assistance, reagents, and critical discussions. We thank G. Petrosillo and G. Paradies (University of Bari, Italy) for their advice and discussion concerning RP-HPLC determination of CL and cyt. c levels. The work was supported by NIHDDK 64166 and NIHGM71520 to MAF, a fellowship award to SYC provided by the United Mitochondrial Disease Foundation, an NRSA T32 fellowship to GMJ, and by AFM (Association Française contre les Myopathies) grant no.11557 (2005) and support from the CNRS (Centre National de la Recherche Scientifique) to PXP.
While this manuscript was under review, the identification of the hCLS gene product was reported elsewhere: Houtkooper et al. 2006. FEBS Lett. 580: 3059–3064; Lu et al. 2006. J Lipid Res. 47: 1140–1145; Chen et al. (2006). Biochem. J. in press.
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Choi, SY., Gonzalvez, F., Jenkins, G. et al. Cardiolipin deficiency releases cytochrome c from the inner mitochondrial membrane and accelerates stimuli-elicited apoptosis. Cell Death Differ 14, 597–606 (2007). https://doi.org/10.1038/sj.cdd.4402020
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DOI: https://doi.org/10.1038/sj.cdd.4402020
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