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TMEM70: a mutational hot spot in nuclear ATP synthase deficiency with a pivotal role in complex V biogenesis

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Abstract

Mammalian complex V (F1F0-ATP synthase or ATPase) uses the proton gradient to generate ATP during oxidative phosphorylation and requires several helper proteins, including TMEM70, to form the holoenzyme in a stepwise process in which nuclear DNA is combined with mitochondrial DNA-encoded subunits. We report the clinical and molecular findings in three patients presenting lactic acidosis, 3-methylglutaconic aciduria, and hypertrophic cardiomyopathy. All three showed an isolated defect of fully assembled ATP synthase in association with a “common” (c.317-2A > G) and a new (c.628A > C/p.T210P) variant in TMEM70. Interestingly, one of the patients also showed nitric oxide-responsive pulmonary arterial hypertension, a finding never before associated with TMEM70 deficiency. In addition to widening the clinical and mutational spectrum of defective ATP synthase, our study also suggests that mutant TMEM70 associates in high molecular weight complexes (470–550 kDa) when expressed in Hela cells and exerts a direct action in ATP synthase biogenesis and assembly, mediating the incorporation of F1 moieties.

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References

  1. Collinson IR, Runswick MJ, Buchanan SK, Fearnley IM, Skehel JM, van Raaij MJ, Griffiths DE, Walker JE (1994) Fo membrane domain of ATP synthase from bovine heart mitochondria: purification, subunit composition, and reconstitution with F1-ATPase. Biochemistry 33:7971–1978

    Article  PubMed  CAS  Google Scholar 

  2. Walker JE, Collinson IR, Van Raaij MJ, Runswick MJ (1995) Structural analysis of ATP synthase from bovine heart mitochondria. Methods Enzymol 260:163–190

    Article  PubMed  CAS  Google Scholar 

  3. Stock D, Leslie AG, Walker JE (1999) Molecular architecture of the rotary motor in ATP synthase. Science 286:1700–1705

    Article  PubMed  CAS  Google Scholar 

  4. Pícková A, Potocký M, Houstek J (2005) Assembly factors of F1FO-ATP synthase across genomes. Proteins 59:393–402

    Article  PubMed  Google Scholar 

  5. Wang ZG, White PS, Ackerman SH (2001) Atp11p and Atp12p are assembly factors for the F(1)-ATPase in human mitochondria. J Biol Chem 276:30773–30778

    Article  PubMed  CAS  Google Scholar 

  6. Cízková A, Stránecký V, Mayr JA, Tesarová M, Havlícková V, Paul J, Ivánek R, Kuss AW, Hansíková H, Kaplanová V, Vrbacký M, Hartmannová H, Nosková L, Honzík T, Drahota Z, Magner M, Hejzlarová K, Sperl W, Zeman J, Houstek J, Kmoch S (2008) TMEM70 mutations cause isolated ATP synthase deficiency and neonatal mitochondrial encephalocardiomyopathy. Nat Genet 40:1288–1290

    Article  PubMed  Google Scholar 

  7. Houstek J, Kmoch S, Zeman J (2009) TMEM70 protein—a novel ancillary factor of mammalian ATP synthase. Biochim Biophys Acta 1787:529–532

    Article  PubMed  CAS  Google Scholar 

  8. Jonckheere AI, Huigsloot M, Lammens M, Jansen J, van den Heuvel LP, Spiekerkoetter U, von Kleist-Retzow JC, Forkink M, Koopman WJ, Szklarczyk R, Huynen MA, Fransen JA, Smeitink JA, Rodenburg RJ (2011) Restoration of complex V deficiency caused by a novel deletion in the human TMEM70 gene normalizes mitochondrial morphology. Mitochondrion 11:954–963

    Article  PubMed  CAS  Google Scholar 

  9. Kucharczyk R, Rak M, di Rago JP (2009) Biochemical consequences in yeast of the human mitochondrial DNA 8993 T > C mutation in the ATPase6 gene found in NARP/MILS patients. Biochim Biophys Acta 1793:817–824

    Article  PubMed  CAS  Google Scholar 

  10. Kucharczyk R, Salin B, di Rago JP (2009) Introducing the human Leigh syndrome mutation T9176G into Saccharomyces cerevisiae mitochondrial DNA leads to severe defects in the incorporation of Atp6p into the ATP synthase and in the mitochondrial morphology. Hum Mol Genet 18:2889–2898

    Article  PubMed  CAS  Google Scholar 

  11. Kucharczyk R, Zick M, Bietenhader M et al (2009) Mitochondrial ATP synthase disorders: molecular mechanisms and the quest for curative therapeutic approaches. Biochim Biophys Acta 1793:186–199

    Article  PubMed  CAS  Google Scholar 

  12. Nijtmans LG, Klement P, Houstĕk J, van den Bogert C (1995) Assembly of mitochondrial ATP synthase in cultured human cells: implications for mitochondrial diseases. Biochim Biophys Acta 1272:190–198

    Article  PubMed  Google Scholar 

  13. Rak M, Gokova S, Tzagoloff A (2011) Modular assembly of yeast mitochondrial ATP synthase. EMBO J 30:920–930

    Article  PubMed  CAS  Google Scholar 

  14. Rak M, Tzagoloff A (2009) F1-dependent translation of mitochondrially encoded Atp6p and Atp8p subunits of yeast ATP synthase. Proc Natl Acad Sci USA 106:18509–18514

    Article  PubMed  CAS  Google Scholar 

  15. Schägger H, Pfeiffer K (2000) Supercomplexes in the respiratory chains of yeast and mammalian mitochondria. EMBO J 19:1777–1783

    Article  PubMed  Google Scholar 

  16. Krause F, Reifschneider NH, Goto S, Dencher NA (2005) Active oligomeric ATP synthases in mammalian mitochondria. Biochem Biophys Res Commun 329:583–590

    Article  PubMed  CAS  Google Scholar 

  17. Jonckheere AI, Smeitink JA, Rodenburg RJ (2012) Mitochondrial ATP synthase: architecture, function and pathology. J Inherit Metab Dis 35:211–225

    Article  PubMed  CAS  Google Scholar 

  18. Hejzlarová K, Tesařová M, Vrbacká-Čížková A, Vrbacký M, Hartmannová H, Kaplanová V, Nosková L, Kratochvílová H, Buzková J, Havlíčková V, Zeman J, Kmoch S, Houštěk J (2011) Expression and processing of the TMEM70 protein. Biochim Biophys Acta 1807:144–149

    Article  PubMed  Google Scholar 

  19. Holt IJ, Harding AE, Petty RK, Morgan-Hughes JA (1990) A new mitochondrial disease associated with mitochondrial DNA heteroplasmy. Am J Hum Genet 46:428–433

    PubMed  CAS  Google Scholar 

  20. Rahman S, Blok RB, Dahl HH, Danks DM, Kirby DM, Chow CW, Christodoulou J, Thorburn DR (1996) Leigh syndrome: clinical features and biochemical and DNA abnormalities. Ann Neurol 39:343–351

    Article  PubMed  CAS  Google Scholar 

  21. Jonckheere AI, Hogeveen M, Nijtmans LG, van den Brand MA, Janssen AJ, Diepstra JH, van den Brandt FC, van den Heuvel LP, Hol FA, Hofste TG, Kapusta L, Dillmann U, Shamdeen MG, Smeitink JA, Rodenburg RJ (2008) A novel mitochondrial ATP8 gene mutation in a patient with apical hypertrophic cardiomyopathy and neuropathy. J Med Genet 45:129–133

    Article  PubMed  CAS  Google Scholar 

  22. Houstek J, Klement P, Floryk D, Antonická H, Hermanská J, Kalous M, Hansíková H, Hout'ková H, Chowdhury SK, Rosipal T, Kmoch S, Stratilová L, Zeman J (1999) A novel deficiency of mitochondrial ATPase of nuclear origin. Hum Mol Genet 8:1967–1974

    Article  PubMed  CAS  Google Scholar 

  23. Houstek J, Mrácek T, Vojtísková A, Zeman J (2004) Mitochondrial diseases and ATPase defects of nuclear origin. Biochim Biophys Acta 1658:115–121

    Article  PubMed  CAS  Google Scholar 

  24. Sperl W, Jesina P, Zeman J, Mayr JA, Demeirleir L, VanCoster R, Pícková A, Hansíková H, Houst'ková H, Krejcík Z, Koch J, Smet J, Muss W, Holme E, Houstek J (2006) Deficiency of mitochondrial ATP synthase of nuclear genetic origin. Neuromuscul Disord 16:821–829

    Article  PubMed  CAS  Google Scholar 

  25. De Meirleir L, Seneca S, Lissens W, De Clercq I, Eyskens F, Gerlo E, Smet J, Van Coster R (2004) Respiratory chain complex V deficiency due to a mutation in the assembly gene ATP12. J Med Genet 41:120–124

    Article  PubMed  Google Scholar 

  26. Shchelochkov OA, Li FY, Wang J, Zhan H, Towbin JA, Jefferies JL, Wong LJ, Scaglia F (2010) Milder clinical course of type IV 3-methylglutaconic aciduria due to a novel mutation in TMEM70. Mol Genet Metab 101:282–285

    Article  PubMed  CAS  Google Scholar 

  27. Spiegel R, Khayat M, Shalev SA, Horovitz Y, Mandel H, Hershkovitz E, Barghuti F, Shaag A, Saada A, Korman SH, Elpeleg O, Yatsiv I (2011) TMEM70 mutations are a common cause of nuclear encoded ATP synthase assembly defect: further delineation of a new syndrome. J Med Genet 48:177–182

    Article  PubMed  Google Scholar 

  28. Tort F, Del Toro M, Lissens W, Montoya J, Fernàndez-Burriel M, Font A, Buján N, Navarro-Sastre A, López-Gallardo E, Arranz JA, Riudor E, Briones P, Ribes A (2011) Screening for nuclear genetic defects in the ATP synthase-associated genes TMEM70, ATP12 and ATP5E in patients with 3-methylglutaconic aciduria. Clin Genet 80:297–300

    Article  PubMed  CAS  Google Scholar 

  29. Mayr JA, Havlícková V, Zimmermann F, Magler I, Kaplanová V, Jesina P, Pecinová A, Nusková H, Koch J, Sperl W, Houstek J (2010) Mitochondrial ATP synthase deficiency due to a mutation in the ATP5E gene for the F1 epsilon subunit. Hum Mol Genet 19:3430–3439

    Article  PubMed  CAS  Google Scholar 

  30. Di Rosa G, Deodato F, Loupatty FJ, Rizzo C, Carrozzo R, Santorelli FM, Boenzi S, D'Amico A, Tozzi G, Bertini E, Maiorana A, Wanders RJ, Dionisi-Vici C (2006) Hypertrophic cardiomyopathy, cataract, developmental delay, lactic acidosis: a novel subtype of 3-methylglutaconic aciduria. J Inherit Metab Dis 29:546–550

    Article  PubMed  Google Scholar 

  31. Rizza T, Vazquez-Memije ME, Meschini MC, Bianchi M, Tozzi G, Nesti C, Piemonte F, Bertini E, Santorelli FM, Carrozzo R (2009) Assaying ATP synthesis in cultured cells: a valuable tool for the diagnosis of patients with mitochondrial disorders. Biochem Biophys Res Commun 383:58–62

    Article  PubMed  CAS  Google Scholar 

  32. Vazquez-Memije ME, Shanske S, Santorelli FM, Kranz-Eble P, Davidson E, DeVivo DC, DiMauro S (1996) Comparative biochemical studies in fibroblasts from patients with different forms of Leigh syndrome. J Inherit Metab Dis 19:43–50

    Article  PubMed  CAS  Google Scholar 

  33. Nijtmans LG, Henderson NS, Holt IJ (2002) Blue Native electrophoresis to study mitochondrial and other protein complexes. Methods 26:327–334

    Article  PubMed  CAS  Google Scholar 

  34. Zerbetto E, Vergani L, Dabbeni-Sala F (1997) Quantification of muscle mitochondrial oxidative phosphorylation enzymes via histochemical staining of blue native polyacrylamide gels. Electrophoresis 18:2059–2064

    Article  PubMed  CAS  Google Scholar 

  35. Carrozzo R, Wittig I, Santorelli FM, Bertini E, Hofmann S, Brandt U, Schägger H (2006) Subcomplexes of human ATP synthase mark mitochondrial biosynthesis disorders. Ann Neurol 59:265–275

    Article  PubMed  CAS  Google Scholar 

  36. Wittig I, Meyer B, Heide H, Steger M, Bleier L, Wumaier Z, Karas M, Schägger H (2010) Assembly and oligomerization of human ATP synthase lacking mitochondrial subunits a and A6L. Biochim Biophys Acta 1797:1004–1011

    Article  PubMed  CAS  Google Scholar 

  37. Ueno M, Murakami T, Takeda A, Kubota M (2009) Efficacy of oral sildenafil in a beraprost-treated patient with severe pulmonary hypertension secondary to type I glycogen storage disease. Circ J 73:1965–1968

    Article  PubMed  Google Scholar 

  38. Leal GN, de Paula AC, Leone C, Kim CA (2010) Echocardiographic study of paediatric patients with mucopolysaccharidosis. Cardiol Young 20:254–261

    Article  PubMed  Google Scholar 

  39. Chan D, Li AM, Yam MC, Li CK, Fok TF (2003) Hurler's syndrome with cor pulmonale secondary to obstructive sleep apnoea treated by continuous positive airway pressure. J Paediatr Child Health 39:558–559

    Article  PubMed  CAS  Google Scholar 

  40. Lo SM, Liu J, Chen F, Pastores GM, Knowles J, Boxer M, Aleck K, Mistry PK (2011) Pulmonary vascular disease in Gaucher disease: clinical spectrum, determinants of phenotype and long-term outcomes of therapy. J Inherit Metab Dis 34:643–650

    Article  PubMed  Google Scholar 

  41. Kovacevic A, Schranz D, Meissner T, Pillekamp F, Schmidt KG (2011) Mucolipidosis II complicated by severe pulmonary hypertension. Mol Genet Metab 104:192–193

    Article  PubMed  CAS  Google Scholar 

  42. Cataltepe S, van Marter LJ, Kozakewich H, Wessel DL, Lee PJ, Levy HL (2000) Pulmonary hypertension associated with nonketotic hyperglycinaemia. J Inherit Metab Dis 23:137–144

    Article  PubMed  CAS  Google Scholar 

  43. Venditti CP, Harris MC, Huff D, Peterside I, Munson D, Weber HS, Rome J, Kaye EM, Shanske S, Sacconi S, Tay S, DiMauro S, Berry GT (2004) Congenital cardiomyopathy and pulmonary hypertension: another fatal variant of cytochrome-c oxidase deficiency. J Inherit Metab Dis 27:735–739

    Article  PubMed  CAS  Google Scholar 

  44. Navarro-Sastre A, Tort F, Stehling O, Uzarska MA, Arranz JA, Del Toro M, Labayru MT, Landa J, Font A, Garcia-Villoria J, Merinero B, Ugarte M, Gutierrez-Solana LG, Campistol A, Garcia-Cazorla J, Vaquerizo E, Riudor P, Briones O, Elpeleg A, Ribes R, Lill J (2011) A fatal mitochondrial disease is associated with defective NFU1 function in the maturation of a subset of mitochondrial Fe-S proteins. Am J Hum Genet 89:656–667

    Article  PubMed  CAS  Google Scholar 

  45. Belostotsky R, Ben-Shalom E, Rinat C, Becker-Cohen R, Feinstein S, Zeligson S, Segel R, Elpeleg O, Nassar S, Frishberg Y (2011) Mutations in the mitochondrial seryl-tRNA synthetase cause hyperuricemia, pulmonary hypertension, renal failure in infancy and alkalosis, HUPRA syndrome. Am J Hum Genet 88:193–200

    Article  PubMed  CAS  Google Scholar 

  46. Schermuly RT, Ghofrani HA, Wilkins MR, Grimminger F (2011) Mechanisms of disease: pulmonary arterial hypertension. Nat Rev Cardiol 8:443–445

    Article  PubMed  CAS  Google Scholar 

  47. Vazquez-Memije ME, Rizza T, Meschini MC, Nesti C, Santorelli FM, Carrozzo R (2009) Cellular and functional analysis of four mutations located in the mitochondrial ATPase6 gene. J Cell Biochem 106:878–886

    Article  PubMed  CAS  Google Scholar 

  48. Cameron JM, Levandovskiy V, Mackay N, Ackerley C, Chitayat D, Raiman J, Halliday WH, Schulze A, Robinson BH (2011) Complex V TMEM70 deficiency results in mitochondrial nucleoid disorganization. Mitochondrion 11:191–199

    Article  PubMed  CAS  Google Scholar 

  49. Havlíčková-Karbanová V, Cížková-Vrbacká A, Hejzlarová K, Nůsková H, Stránecký V, Potocká A, Kmoch S, Houštěk J (2012) Compensatory upregulation of respiratory chain complexes III and IV in isolated deficiency of ATP synthase due to TMEM70 mutation. Biochim Biophys Acta 1817(7):1037–1043. doi:10.1016/j.bbabio.2012.03.004

    Article  PubMed  Google Scholar 

  50. Manczak M, Jung Y, Park BS, Partovi D, Reddy PH (2005) Time-course of mitochondrial gene expressions in mice brains: implications for mitochondrial dysfunction, oxidative damage, and cytochrome c in aging. J Neurochem 92:494–504

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors thank Dr. S. Petrini for providing technical assistance with the confocal microscopy for immunocytochemistry of Hela cells. This research was supported by grants from the Telethon-Mitocon Project (Telethon GUP09004) and from the FP7 LeukoTreat European Project; it was also supported by the Italian Health Ministry through “Ricerca Finalizzata” funding to EB and “Ricerca Corrente” funding to RC. Dr. Diego Martinelli is supported by a fellowship from the association “La vita è un Dono”. The authors acknowledge the editorial assistance of Mrs. Catherine J. Wrenn.

Conflict of interest

The authors declare that they have no conflict of interest.

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Author notes

  1. Maria Chiara Meschini

    Present address: UOC Neurogenetica e Malattie Neuromuscolari, IRCCS Fondazione Stella Maris, Pisa, Italy

Authors and Affiliations

  1. Unit of Molecular Medicine for Neuromuscular and Neurodegenerative Diseases, Bambino Gesù Children’s Hospital, IRCCS, Rome, Italy

    Alessandra Torraco, Daniela Verrigni, Teresa Rizza, Maria Chiara Meschini, Marzia Bianchi, Fiorella Piemonte, Enrico Bertini & Rosalba Carrozzo

  2. Unidad de Investigación Médica en Génetica, Centro Médico Nacional S.XXI, Instituto Mexicano del Seguro Social, Mexico City, Mexico

    Martha Elisa Vazquez-Memije

  3. Division of Metabolism, Bambino Gesù Children’s Hospital, IRCCS, Rome, Italy

    Diego Martinelli & Carlo Dionisi-Vici

  4. UOC Neurogenetica e Malattie Neuromuscolari, IRCCS Fondazione Stella Maris, Pisa, Italy

    Filippo Maria Santorelli

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  1. Alessandra Torraco
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Correspondence to Rosalba Carrozzo.

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Alessandra Torraco, Daniela Verrigni, and Teresa Rizza contributed equally to this work.

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Torraco, A., Verrigni, D., Rizza, T. et al. TMEM70: a mutational hot spot in nuclear ATP synthase deficiency with a pivotal role in complex V biogenesis. Neurogenetics 13, 375–386 (2012). https://doi.org/10.1007/s10048-012-0343-8

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