Mitochondrial calcium uniporter protein MCU is involved in oxidative stress-induced cell death

Abstract

Mitochondrial calcium uniporter (MCU) is a conserved Ca²⁺ transporter at mitochondrial in eukaryotic cells. However, the role of MCU protein in oxidative stressinduced cell death remains unclear. Here, we showed that ectopically expressed MCU is mitochondrial localized in both HeLa and primary cerebellar granule neurons (CGNs). Knockdown of endogenous MCU decreases mitochondrial Ca²⁺ uptake following histamine stimulation and attenuates cell death induced by oxidative stress in both HeLa cells and CGNs. We also found MCU interacts with VDAC1 and mediates VDAC1 overexpression-induced cell death in CGNs. This finding demonstrates that MCU-VDAC1 complex regulates mitochondrial Ca²⁺ uptake and oxidative stress-induced apoptosis, which might represent therapeutic targets for oxidative stress related diseases.

Keywords

MCU / VDAC1 / oxidative stress / calcium uptake / cell death

References

[1] Ahuja M, Muallem S (2014) The gatekeepers of mitochondrial calcium influx: MICU1 and MICU2. EMBO Rep15: 205-206

[2] Alam MR, Groschner LN, Parichatikanond W, Kuo L, Bondarenko AI, Rost R, Waldeck-Weiermair M, Malli R, Graier WF (2012) Mitochondrial Ca²⁺ uptake 1 (MICU1) and mitochondrial Ca²⁺ uniporter (MCU) contribute to metabolism-secretion coupling in clonal pancreatic beta-cells. J Biol Chem287: 34445-34454

[3] Baughman JM, Perocchi F, Girgis HS, Plovanich M, Belcher-Timme CA, Sancak Y, Bao XR, Strittmatter L, Goldberger O, Bogorad RL, Koteliansky V, Mootha VK (2011) Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Nature476: 341-345

[4] Ben-Hail D, Shoshan-Barmatz V (2012) Anion-transport blockers inhibit VDAC1 channel conductance, oligomerization and apoptosis. Febs J279: 429-429

[5] Brustovetsky N, Brustovetsky T, Jemmerson R, Dubinsky JM (2002) Calcium-induced cytochrome c release from CNS mitochondria is associated with the permeability transition and rupture of the outer membrane. J Neurochem80: 207-218

[6] Chu ZL, Pio F, Xie Z, Welsh K, Krajewska M, Krajewski S, Godzik A, Reed JC (2001) A novel enhancer of the Apaf1 apoptosome involved in cytochrome c-dependent caspase activation and apoptosis. J Biol Chem276: 9239-9245

[7] De Stefani D, Raffaello A, Teardo E, Szabo I, Rizzuto R (2011) A forty-kilodalton protein of the inner membrane is the mitochondrial calcium uniporter. Nature476: 336-340

[8] Dong HL, Wang SL, Zhang ZW, Yu AL, Liu Z (2014) The effect of mitochondrial calcium uniporter opener spermine on diazoxide against focal cerebral ischemia–reperfusion injury in rats. J Stroke Cerebrovasc23: 303-309

[9] Fernandez-Echevarria C, Diaz M, Ferrer I, Canerina-Amaro A, Marin R (2014) Abeta promotes VDAC1 channel dephosphorylation in neuronal lipid rafts. Relevance to the mechanisms of neurotoxicity in Alzheimer’s disease. Neuroscience278: 354-366

[10] Gincel D, Zaid H, Shoshan-Barmatz V (2001) Calcium binding and translocation by the voltage-dependent anion channel: a possible regulatory mechanism in mitochondrial function. Biochem J358: 147-155

[11] Halestrap AP, Connern CP, Griffiths EJ, Kerr PM (1997) Cyclosporin A binding to mitochondrial cyclophilin inhibits the permeability transition pore and protects hearts from ischaemia/reperfusion injury. Mol Cell Biochem174: 167-172

[12] Hall DD, Wu YJ, Domann FE, Spitz DR, Anderson ME (2014) Mitochondrial calcium uniporter activity is dispensable for MDAMB- 231 breast carcinoma cell survival. PLoS ONE9: e96866

[13] Keinan N, Pahima H, Ben-Hail D, Shoshan-Barmatz V (2013) The role of calcium in VDAC1 oligomerization and mitochondriamediated apoptosis. Bba-Mol Cell Res1833: 1745-1754

[14] Kinnally KW, Peixoto PM, Ryu SY, Dejean LM (2011) Is mPTP the gatekeeper for necrosis, apoptosis, or both? Biochimica et biophysica acta1813: 616-622

[15] Konishi Y, Bonni A (2003) The E2F–Cdc2 cell-cycle pathway specifically mediates activity deprivation-induced apoptosis of postmitotic neurons. J Neurosci23: 1649-1658

[16] Kumfu S, Chattipakorn S, Fucharoen S, Chattipakorn N (2012) Mitochondrial calcium uniporter blocker prevents cardiac mitochondrial dysfunction induced by iron overload in thalassemic mice. Biometals25: 1167-1175

[17] Madesh M, Hajnoczky G (2001) VDAC-dependent permeabilization of the outer mitochondrial membrane by superoxide induces rapid and massive cytochrome c release. J Cell Biol155: 1003-1015

[18] Mallilankaraman K, Cardenas C, Doonan PJ, Chandramoorthy HC, Irrinki KM, Golenar T, Csordas G, Madireddi P, Yang J, Muller M, Miller R, Kolesar JE, Molgo J, Kaufman B, Hajnoczky G, Foskett JK, Madesh M (2012a) MCUR1 is an essential component of mitochondrial Ca²⁺ uptake that regulates cellular metabolism. Nature Cell Biol14: 1336-1343

[19] Mallilankaraman K, Doonan P, Cardenas C, Chandramoorthy HC, Muller M, Miller R, Hoffman NE, Gandhirajan RK, Molgo J, Birnbaum MJ, Rothberg BS, Mak DO, Foskett JK, Madesh M (2012b) MICU1 is an essential gatekeeper for MCU-mediated mitochondrial Ca(2+) uptake that regulates cell survival. Cell151: 630-644

[20] Naranmandura H, Chen X, Tanaka M, Wang WW, Rehman K, Xu S, Chen Z, Chen SQ, Suzuki N (2012) Release of apoptotic cytochrome C from mitochondria by dimethylarsinous acid occurs through interaction with voltage-dependent anion channel in vitro. Toxicol Sci128: 137-146

[21] Pan X, Liu J, Nguyen T, Liu CY, Sun JH, Teng YJ, Fergusson MM, Rovira II, Allen M, Springer DA, Aponte AM, Gucek M, Balaban RS, Murphy E, Finkel T (2013) The physiological role of mitochondrial calcium revealed by mice lacking the mitochondrial calcium uniporter. Nature Cell Biol15: 1464-1472

[22] Pestana CR, Silva CH, Uyemura SA, Santos AC, Curti C (2010) Impact of adenosine nucleotide translocase (ANT) proline isomerization on Ca²⁺-induced cysteine relative mobility/mitochondrial permeability transition pore. J Bioenerg Biomembr42: 329-335

[23] Qiu J, Tan YW, Hagenston AM, Martel MA, Kneisel N, Skehel PA, Wyllie DJ, Bading H, Hardingham GE (2013) Mitochondrial calcium uniporter MCU controls excitotoxicity and is transcriptionally repressed by neuroprotective nuclear calcium signals. Nat Commun4: 2034

[24] Raffaello A, De Stefani D, Sabbadin D, Teardo E, Merli G, Picard A, Checchetto V, Moro S, Szabo I, Rizzuto R (2013) The mitochondrial calcium uniporter is a multimer that can include a dominantnegative pore-forming subunit. EMBO J32: 2362-2376

[25] Rapizzi E, Pinton P, Szabadkai G, Wieckowski MR, Vandecasteele G, Baird G, Tuft RA, Fogarty KE, Rizzuto R (2002) Recombinant expression of the voltage-dependent anion channel enhances the transfer of Ca2+ microdomains to mitochondria. J Cell Biol159: 613-624

[26] Robb-Gaspers LD, Burnett P, Rutter GA, Denton RM, Rizzuto R, Thomas AP (1998) Integrating cytosolic calcium signals into mitochondrial metabolic responses. Embo J17: 4987-5000

[27] Sancak Y, Markhard AL, Kitami T, Kovacs-Bogdan E, Kamer KJ, Udeshi ND, Carr SA, Chaudhuri D, Clapham DE, Li AA, Calvo SE, Goldberger O, Mootha VK (2013) EMRE is an essential component of the mitochondrial calcium uniporter complex. Science342: 1379-1382

[28] Schwartz J, Holmuhamedov E, Zhang X, Lovelace GL, Smith CD, Lemasters JJ (2013) Minocycline and doxycycline, but not other tetracycline-derived compounds, protect liver cells from chemical hypoxia and ischemia/reperfusion injury by inhibition of the mitochondrial calcium uniporter. Toxicol Appl Pharmacol273: 172-179

[29] Shoshan-Barmatz V, Zakar M, Rosenthal K, Abu-Hamad S (2009) Key regions of VDAC1 functioning in apoptosis induction and regulation by hexokinase. Biochimica et biophysica acta1787: 421-430

[30] Shoshan-Barmatz V, Keinan N, Abu-Hamad S, Tyomkin D, Aram L (2010) Apoptosis is regulated by the VDAC1 N-terminal region and by VDAC oligomerization: release of cytochrome c, AIF and Smac/Diablo. Bba-Bioenergetics1797: 1281-1291

[31] Sripetchwandee J, Kenknight S, Sanit J, Chattipakorn S, Chattipakorn N (2013a) Blocking mitochondrial calcium uniporter completely prevents cardiac mitochondrial dysfunction caused by iron overload. J Am Coll Cardiol61: E705-E705

[32] Sripetchwandee J, Sanit J, Chattipakorn N, Chattipakorn SC (2013b) Mitochondrial calcium uniporter blocker effectively prevents brain mitochondrial dysfunction caused by iron overload. Life Sci92: 298-304

[33] Sripetchwandee J, KenKnight SB, Sanit J, Chattipakorn S, Chattipakorn N (2014) Blockade of mitochondrial calcium uniporter prevents cardiac mitochondrial dysfunction caused by iron overload. Acta Physiol210: 330-341

[34] Xie Q, Hao Y, Tao L, Peng S, Rao C, Chen H, You H, Dong MQ, Yuan Z (2012) Lysine methylation of FOXO3 regulates oxidative stress-induced neuronal cell death. EMBO reports13: 371-377

[35] Xu X, Decker W, Sampson MJ, Craigen WJ, Colombini M (1999) Mouse VDAC isoforms expressed in yeast: channel properties and their roles in mitochondrial outer membrane permeability. J Membr Biol170: 89-102

[36] Zhao Q, Wang SL, Li Y, Wang P, Li SH, Guo YL, Yao RY (2013) The role of the mitochondrial calcium uniporter in cerebral ischemia/ reperfusion injury in rats involves regulation of mitochondrial energy metabolism. Mol Med Rep7: 1073-1080

[37] Zheng Y, Shi Y, Tian C, Jiang C, Jin H, Chen J, Almasan A, Tang H, Chen Q (2004) Essential role of the voltage-dependent anion channel (VDAC) in mitochondrial permeability transition pore opening and cytochrome c release induced by arsenic trioxide. Oncogene23: 1239-1247

RIGHTS & PERMISSIONS

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.