Acetyl-l-carnitine protects yeast cells from apoptosis and aging and inhibits mitochondrial fission
Vanessa Palermo
Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, University of Rome “La Sapienza” Piazzale Aldo Moro, Rome, Italy
Search for more papers by this authorClaudio Falcone
Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, University of Rome “La Sapienza” Piazzale Aldo Moro, Rome, Italy
Search for more papers by this authorMenotti Calvani
Scientific Department, Sigma-Tau, Via Pontina Km. Pomezia, Rome, Italy
Search for more papers by this authorCristina Mazzoni
Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, University of Rome “La Sapienza” Piazzale Aldo Moro, Rome, Italy
Search for more papers by this authorVanessa Palermo
Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, University of Rome “La Sapienza” Piazzale Aldo Moro, Rome, Italy
Search for more papers by this authorClaudio Falcone
Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, University of Rome “La Sapienza” Piazzale Aldo Moro, Rome, Italy
Search for more papers by this authorMenotti Calvani
Scientific Department, Sigma-Tau, Via Pontina Km. Pomezia, Rome, Italy
Search for more papers by this authorCristina Mazzoni
Department of Cell and Developmental Biology, Pasteur Institute-Cenci Bolognetti Foundation, University of Rome “La Sapienza” Piazzale Aldo Moro, Rome, Italy
Search for more papers by this authorSummary
In this work we report that carnitines, in particular acetyl-l-carnitine (ALC), are able to prolong the chronological aging of yeast cells during the stationary phase. Lifespan extension is significantly reduced in yca1 mutants as well in rho0 strains, suggesting that the protective effects pass through the Yca1 caspase and mitochondrial functions. ALC can also prevent apoptosis in pro-apoptotic mutants, pointing to the importance of mitochondrial functions in regulating yeast apoptosis and aging. We also demonstrate that ALC attenuates mitochondrial fission in aged yeast cells, indicating a correlation between its protective effect and this process. Our findings suggest that ALC, used as therapeutic for stroke, myocardial infarction and neurodegenerative diseases, besides the well-known anti-oxidant effects, might exert protective effects also acting on mitochondrial morphology.
Supporting Information
Fig. S1 CML39-11A (A) and MCY4/313Kllsm4Δ1 (lsm4Δ1, B) were grown in 0.2% glucose. Respiration rates are expressed as nanomoles of oxygen consumed per milliliter of solution per minute. Average and standard deviations, obtained from three independent experiments, are also indicated.
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References
- Alirol E, Martinou JC (2006) Mitochondria and cancer: is there a morphological connection? Oncogene 25, 4706–4716.
- Balaban RS, Nemoto S, Finkel T (2005) Mitochondria, oxidants, and aging. Cell 120, 483–495.
- Binienda Z, Przybyla-Zawislak B, Virmani A, Schmued L (2005) L-carnitine and neuroprotection in the animal model of mitochondrial dysfunction. Ann. N Y Acad. Sci. 1053, 174–182.
- Bossy-Wetzel E, Barsoum MJ, Godzik A, Schwarzenbacher R, Lipton SA (2003) Mitochondrial fission in apoptosis, neurodegeneration and aging. Curr. Opin. Cell Biol. 15, 706–716.
-
Brachmann CB,
Davies A,
Cost GJ,
Caputo E,
Li J,
Hieter P,
Boeke JD (1998) Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications.
Yeast
14, 115–132.
10.1002/(SICI)1097-0061(19980130)14:2<115::AID-YEA204>3.0.CO;2-2 CAS PubMed Web of Science® Google Scholar
- Buttner S, Eisenberg T, Carmona-Gutierrez D, Ruli D, Knauer H, Ruckenstuhl C, Sigrist C, Wissing S, Kollroser M, Frohlich KU, Sigrist S, Madeo F (2007) Endonuclease G regulates budding yeast life and death. Mol. Cell 25, 233–246.
- Calabrese V, Cornelius C, Mancuso C, Pennisi G, Calafato S, Bellia F, Bates TE, Giuffrida Stella AM, Schapira T, Dinkova Kostova AT, Rizzarelli E (2008) Cellular stress response: a novel target for chemoprevention and nutritional neuroprotection in aging, neurodegenerative disorders and longevity. Neurochem. Res. 33, 2444–2471.
- Calvani M, Benatti P, Mancinelli A, D’Iddio S, Giordano V, Koverech A, Amato A, Brass EP (2004) Carnitine replacement in end-stage renal disease and hemodialysis. Ann. N Y Acad. Sci. 1033, 52–66.
- Cassidy-Stone A, Chipuk JE, Ingerman E, Song C, Yoo C, Kuwana T, Kurth MJ, Shaw JT, Hinshaw JE, Green DR, Nunnari J (2008) Chemical inhibition of the mitochondrial division dynamin reveals its role in Bax/Bak-dependent mitochondrial outer membrane permeabilization. Dev. Cell. 14, 193–204.
- Dhitavat S, Ortiz D, Shea TB, Rivera ER (2002) Acetyl-L-carnitine protects against amyloid-beta neurotoxicity: roles of oxidative buffering and ATP levels. Neurochem. Res. 27, 501–505.
- Escobar-Henriques M, Westermann B, Langer T (2006) Regulation of mitochondrial fusion by the F-box protein Mdm30 involves proteasome-independent turnover of Fzo1. J. Cell Biol. 173, 645–650.
- Fabrizio P, Pozza F, Pletcher SD, Gendron CM, Longo VD (2001) Regulation of longevity and stress resistance by Sch9 in yeast. Science 292, 288–290.
- Fahrenkrog B, Sauder U, Aebi U (2004) The S. cerevisiae HtrA-like protein Nma111p is a nuclear serine protease that mediates yeast apoptosis. J. Cell Sci. 117, 115–126.
- Frank S, Gaume B, Bergmann-Leitner ES, Leitner WW, Robert EG, Catez F, Smith CL, Youle RJ (2001) The role of dynamin-related protein 1, a mediator of mitochondrial fission, in apoptosis. Dev. Cell. 1, 515–525.
- Franken J, Kroppenstedt S, Swiegers JH, Bauer FF (2008) Carnitine and carnitine acetyltransferases in the yeast Saccharomyces cerevisiae: a role for carnitine in stress protection. Curr. Genet. 53, 347–360.
- Goldberg AA, Bourque SD, Kyryakov P, Gregg C, Boukh-Viner T, Beach A, Burstein MT, Machkalyan G, Richard V, Rampersad S, Cyr D, Milijevic S, Titorenko VI (2009) Effect of calorie restriction on the metabolic history of chronologically aging yeast. Exp. Gerontol. 44, 555–571.
- Gulcin I (2006) Antioxidant and antiradical activities of L-carnitine. Life Sci. 78, 803–811.
- Heath-Engel HM, Shore GC (2006) Mitochondrial membrane dynamics, cristae remodelling and apoptosis. Biochim. Biophys. Acta 1763, 549–560.
- Hoppins S, Lackner L, Nunnari J (2007) The machines that divide and fuse mitochondria. Annu. Rev. Biochem. 76, 751–780.
- Huizing M, Iacobazzi V, Ijlst L, Savelkoul P, Ruitenbeek W, Van DenHeuvelL, Indiveri C, Smeitink J, Trijbels F, Wanders R, Palmieri F (1997) Cloning of the human carnitine-acylcarnitine carrier cDNA and identification of the molecular defect in a patient. Am. J. Hum. Genet. 61, 1239–1245.
- Jagasia R, Grote P, Westermann B, Conradt B (2005) DRP-1-mediated mitochondrial fragmentation during EGL-1-induced cell death in C. elegans. Nature 433, 754–760.
- Lee YJ, Jeong SY, Karbowski M, Smith CL, Youle RJ (2004) Roles of the mammalian mitochondrial fission and fusion mediators Fis1, Drp1, and Opa1 in apoptosis. Mol. Biol. Cell 15, 5001–5011.
- Ligr M, Madeo F, Frohlich E, Hilt W, Frohlich KU, Wolf DH (1998) Mammalian Bax triggers apoptotic changes in yeast. FEBS Lett. 438, 61–65.
- Lin SJ, Kaeberlein M, Andalis AA, Sturtz LA, Defossez PA, Culotta VC, Fink GR, Guarente L (2002) Calorie restriction extends Saccharomyces cerevisiae lifespan by increasing respiration. Nature 418, 344–348.
- Madeo F, Frohlich E, Frohlich KU (1997) A yeast mutant showing diagnostic markers of early and late apoptosis. J. Cell Biol. 139, 729–734.
- Madeo F, Frohlich E, Ligr M, Grey M, Sigrist SJ, Wolf DH, Frohlich KU (1999) Oxygen stress: a regulator of apoptosis in yeast. J. Cell Biol. 145, 757–767.
- Madeo F, Herker E, Maldener C, Wissing S, Lachelt S, Herlan M, Fehr M, Lauber K, Sigrist SJ, Wesselborg S, Frohlich KU (2002) A caspase-related protease regulates apoptosis in yeast. Mol. Cell 9, 911–917.
- Mazzio E, Yoon KJ, Soliman KF (2003) Acetyl-L-carnitine cytoprotection against 1-methyl-4-phenylpyridinium toxicity in neuroblastoma cells. Biochem. Pharmacol. 66, 297–306.
- Mazzoni C, Falcone C (2001) Isolation and study of KlLSM4, a Kluyveromyces lactis gene homologous to the essential gene LSM4 of Saccharomyces cerevisiae. Yeast 18, 1249–1256.
- Mazzoni C, Falcone C (2008) Caspase-dependent apoptosis in yeast. Biochim. Biophys. Acta 1783, 1320–1327.
- Mazzoni C, Mancini P, Verdone L, Madeo F, Serafini A, Herker E, Falcone C (2003) A truncated form of KlLsm4p and the absence of factors involved in mRNA decapping trigger apoptosis in yeast. Mol. Biol. Cell 14, 721–729.
- Mazzoni C, Herker E, Palermo V, Jungwirth H, Eisenberg T, Madeo F, Falcone C (2005a) Yeast caspase 1 links messenger RNA stability to apoptosis in yeast. EMBO Rep. 6, 1076–1081.
- Mazzoni C, Palermo V, Torella M, Falcone C (2005b) HIR1, the co-repressor of histone gene transcription of Saccharomyces cerevisiae, acts as a multicopy suppressor of the apoptotic phenotypes of the LSM4 mRNA degradation mutant. FEMS Yeast Res. 5, 1229–1235.
- Mazzoni C, Torella M, Petrera A, Palermo V, Falcone C (2009) PGK1, the gene encoding the glycolitic enzyme phosphoglycerate kinase, acts as a multicopy suppressor of apoptotic phenotypes in S. cerevisiae. Yeast 26, 31–37.
- Mozdy AD, McCaffery JM, Shaw JM (2000) Dnm1p GTPase-mediated mitochondrial fission is a multi-step process requiring the novel integral membrane component Fis1p. J. Cell Biol. 151, 367–380.
- Neuspiel M, Zunino R, Gangaraju S, Rippstein P, McBride H (2005) Activated mitofusin 2 signals mitochondrial fusion, interferes with Bax activation, and reduces susceptibility to radical induced depolarization. J. Biol. Chem. 280, 25060–25070.
- Olichon A, Baricault L, Gas N, Guillou E, Valette A, Belenguer P, Lenaers G (2003) Loss of OPA1 perturbates the mitochondrial inner membrane structure and integrity, leading to cytochrome c release and apoptosis. J. Biol. Chem. 278, 7743–7746.
- Palermo V, Falcone C, Mazzoni C (2007) Apoptosis and aging in mitochondrial morphology mutants of S. cerevisiae. Folia. Microbiol. (Praha) 52, 479–483.
- Parone PA, Martinou JC (2006) Mitochondrial fission and apoptosis: an ongoing trial. Biochim. Biophys. Acta 1763, 522–530.
- Pillich RT, Scarsella G, Risuleo G (2005) Reduction of apoptosis through the mitochondrial pathway by the administration of acetyl-L-carnitine to mouse fibroblasts in culture. Exp. Cell Res. 306, 1–8.
- Reddy JK, Mannaerts GP (1994) Peroxisomal lipid metabolism. Annu. Rev. Nutr. 14, 343–370.
- Van Roermund CW, Elgersma Y, Singh N, Wanders RJ, Tabak HF (1995) The membrane of peroxisomes in Saccharomyces cerevisiae is impermeable to NAD(H) and acetyl-CoA under in vivo conditions. EMBO J. 14, 3480–3486.
- Schmalix W, Bandlow W (1993) The ethanol-inducible YAT1 gene from yeast encodes a presumptive mitochondrial outer carnitine acetyltransferase. J. Biol. Chem. 268, 27428–27439.
- Scorrano L (2005) Proteins that fuse and fragment mitochondria in apoptosis: con-fissing a deadly con-fusion? J. Bioenerg. Biomembr. 37, 165–170.
- Shirogane T, Fukada T, Muller JM, Shima DT, Hibi M, Hirano T (1999) Synergistic roles for Pim-1 and c-Myc in STAT3-mediated cell cycle progression and antiapoptosis. Immunity 11, 709–719.
- Smith DLJ, McClure JM, Matecic M, Smith JS (2007) Calorie restriction extends the chronological lifespan of Saccharomyces cerevisiae independently of the Sirtuins. Aging Cell 6, 649–662.
- Stemple CJ, Davis MA, Hynes MJ (1998) The facC gene of Aspergillus nidulans encodes an acetate-inducible carnitine acetyltransferase. J. Bacteriol. 180, 6242–6251.
- Sugioka R, Shimizu S, Tsujimoto Y (2004) Fzo1, a protein involved in mitochondrial fusion, inhibits apoptosis. J. Biol. Chem. 279, 52726–52734.
-
Westermann B,
Neupert W (2000) Mitochondria-targeted green fluorescent proteins: convenient tools for the study of organelle biogenesis in Saccharomyces cerevisiae.
Yeast
16, 1421–1427.
10.1002/1097-0061(200011)16:15<1421::AID-YEA624>3.0.CO;2-U CAS PubMed Web of Science® Google Scholar
- Wissing S, Ludovico P, Herker E, Buttner S, Engelhardt SM, Decker T, Link A, Proksch A, Rodrigues F, Corte-Real M, Frohlich KU, Manns J, Cande C, Sigrist SJ, Kroemer G, Madeo F (2004) An AIF orthologue regulates apoptosis in yeast. J. Cell Biol. 166, 969–974.
- Yano T, Itoh Y, Yamada M, Egashira N, Oishi R (2008) Combined treatment with L-carnitine and a pan-caspase inhibitor effectively reverses amiodarone-induced injury in cultured human lung epithelial cells. Apoptosis 13, 543–552.