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Loss of ETHE1, a mitochondrial dioxygenase, causes fatal sulfide toxicity in ethylmalonic encephalopathy

Nature Medicine volume 15, pages 200–205 (2009)Cite this article

An Erratum to this article was published on 01 February 2009

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Abstract

Ethylmalonic encephalopathy is an autosomal recessive, invariably fatal disorder characterized by early-onset encephalopathy, microangiopathy, chronic diarrhea, defective cytochrome c oxidase (COX) in muscle and brain, high concentrations of C4 and C5 acylcarnitines in blood and high excretion of ethylmalonic acid in urine. ETHE1, a gene encoding a β-lactamase–like, iron-coordinating metalloprotein, is mutated in ethylmalonic encephalopathy. In bacteria, ETHE1-like sequences are in the same operon of, or fused with, orthologs of TST, the gene encoding rhodanese, a sulfurtransferase. In eukaryotes, both ETHE1 and rhodanese are located within the mitochondrial matrix. We created a Ethe1−/− mouse that showed the cardinal features of ethylmalonic encephalopathy. We found that thiosulfate was excreted in massive amounts in urine of both Ethe1−/− mice and humans with ethylmalonic encephalopathy. High thiosulfate and sulfide concentrations were present in Ethe1−/− mouse tissues. Sulfide is a powerful inhibitor of COX and short-chain fatty acid oxidation, with vasoactive and vasotoxic effects that explain the microangiopathy in ethylmalonic encephalopathy patients. Sulfide is detoxified by a mitochondrial pathway that includes a sulfur dioxygenase. Sulfur dioxygenase activity was absent in Ethe1−/− mice, whereas it was markedly increased by ETHE1 overexpression in HeLa cells and Escherichia coli. Therefore, ETHE1 is a mitochondrial sulfur dioxygenase involved in catabolism of sulfide that accumulates to toxic levels in ethylmalonic encephalopathy.

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Figure 1: Genotypic and phenotypic analysis of Ethe1 recombinant mice.
Figure 2: Histochemical and biochemical results of MRC activities in mouse tissues.
Figure 3: Metabolic and structural characterization in ETHE1−/− mice and subjects with ethylmalonic encephalopathy.
Figure 4: Sulfur dioxygenase (SDO) activity in mouse liver, human HeLa cells and affinity-purified recombinant ETHE1His from E. coli.

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  • 20 January 2009

    In the version of this article initially published online, ‘SDO’ should have been ‘SDH’ in Figure 4a,b. The error has been corrected for the print, PDF and HTML versions of this article.

References

  1. Burlina, A. et al. New clinical phenotype of branched-chain acyl-CoA oxidation defect. Lancet 338, 1522–1523 (1991).

    Article  CAS  Google Scholar 

  2. Garcia-Silva, M.T. et al. Encephalopathy, petechiae, and acrocyanosis with ethylmalonic aciduria associated with muscle cytochrome c oxidase deficiency. J. Pediatr. 125, 843–844 (1994).

    Article  CAS  Google Scholar 

  3. Gregersen, N. et al. Identification of four new mutations in the short-chain acyl-CoA dehydrogenase (SCAD) gene in two patients: one of the variant alleles, 511C → T, is present at an unexpectedly high frequency in the general population, as was the case for 625G → A, together conferring susceptibility to ethylmalonic aciduria. Hum. Mol. Genet. 7, 619–627 (1998).

    Article  CAS  Google Scholar 

  4. Koeberl, D.D. et al. Rare disorders of metabolism with elevated butyryl- and isobutyryl-carnitine detected by tandem mass spectrometry newborn screening. Pediatr. Res. 54, 219–223 (2003).

    Article  CAS  Google Scholar 

  5. Sass, J.O. et al. 2-Methylbutyryl-coenzyme A dehydrogenase deficiency: functional and molecular studies on a defect in isoleucine catabolism. Mol. Genet. Metab. 93, 30–35 (2008).

    Article  CAS  Google Scholar 

  6. Vockley, J. & Ensenauer, R. Isovaleric acidemia: new aspects of genetic and phenotypic heterogeneity. Am. J. Med. Genet. C. Semin. Med. Genet. 142C, 95–103 (2006).

    Article  CAS  Google Scholar 

  7. Tiranti, V. et al. Ethylmalonic encephalopathy is caused by mutations in ETHE1, a gene encoding a mitochondrial matrix protein. Am. J. Hum. Genet. 74, 239–252 (2004).

    Article  CAS  Google Scholar 

  8. Tiranti, V. et al. ETHE1 mutations are specific to ethylmalonic encephalopathy. J. Med. Genet. 43, 340–346 (2006).

    Article  CAS  Google Scholar 

  9. Mineri, R. et al. Identification of new mutations in the ETHE1 gene in a cohort of 14 patients presenting with ethylmalonic encephalopathy. J. Med. Genet. 45, 473–478 (2008).

    Article  CAS  Google Scholar 

  10. Daiyasu, H., Osaka, K., Ishino, Y. & Toh, H. Expansion of the zinc metallo-hydrolase family of the β-lactamase fold. FEBS Lett. 503, 1–6 (2001).

    Article  CAS  Google Scholar 

  11. Higashitsuji, H. et al. A novel protein overexpressed in hepatoma accelerates export of NF-κ B from the nucleus and inhibits p53-dependent apoptosis. Cancer Cell 2, 335–346 (2002).

    Article  CAS  Google Scholar 

  12. Higashitsuji, H. et al. Enhanced deacetylation of p53 by the anti-apoptotic protein HSCO in association with histone deacetylase 1. J. Biol. Chem. 282, 13716–13725 (2007).

    Article  CAS  Google Scholar 

  13. Bensaad, K. & Vousden, K.H. p53: new roles in metabolism. Trends Cell Biol. 17, 286–291 (2007).

    Article  CAS  Google Scholar 

  14. Psarra, A.M. & Sekeris, C.E. Nuclear receptors and other nuclear transcription factors in mitochondria: regulatory molecules in a new environment. Biochim. Biophys. Acta 1783, 1–11 (2008).

    Article  CAS  Google Scholar 

  15. McCoy, J.G. et al. Structure of an ETHE1-like protein from Arabidopsis thaliana. Acta Crystallogr. D Biol. Crystallogr. 62, 964–970 (2006).

    Article  Google Scholar 

  16. Szabo, C. Hydrogen sulphide and its therapeutic potential. Nat. Rev. Drug Discov. 6, 917–935 (2007).

    Article  CAS  Google Scholar 

  17. Leschelle, X. et al. Adaptative metabolic response of human colonic epithelial cells to the adverse effects of the luminal compound sulfide. Biochim. Biophys. Acta 1725, 201–212 (2005).

    Article  CAS  Google Scholar 

  18. Dorman, D.C. et al. Cytochrome oxidase inhibition induced by acute hydrogen sulfide inhalation: correlation with tissue sulfide concentrations in the rat brain, liver, lung, and nasal epithelium. Toxicol. Sci. 65, 18–25 (2002).

    Article  CAS  Google Scholar 

  19. Garcia-Silva, M.T., Ribes, A., Campos, Y., Garavaglia, B. & Arenas, J.A. Syndrome of Encephalopathy, Petechiae, and Ethylmalonic Aciduria. Pediatric Neurology. 17, 165–170 (1997).

    Article  CAS  Google Scholar 

  20. Furne, J., Springfield, J., Koenig, T., DeMaster, E. & Levitt, M.D. Oxidation of hydrogen sulfide and methanethiol to thiosulfate by rat tissues: a specialized function of the colonic mucosa. Biochem. Pharmacol. 62, 255–259 (2001).

    Article  CAS  Google Scholar 

  21. Roediger, W.E., Moore, J. & Babidge, W. Colonic sulfide in pathogenesis and treatment of ulcerative colitis. Dig. Dis. Sci. 42, 1571–1579 (1997).

    Article  CAS  Google Scholar 

  22. Hildebrandt, T.M. & Grieshaber, M.K. Three enzymatic activities catalyze the oxidation of sulfide to thiosulfate in mammalian and invertebrate mitochondria. FEBS J. 275, 3352–3361 (2008).

    Article  CAS  Google Scholar 

  23. Goubern, M., Andriamihaja, M., Nubel, T., Blachier, F. & Bouillaud, F. Sulfide, the first inorganic substrate for human cells. FASEB J. 21, 1699–1706 (2007).

    Article  CAS  Google Scholar 

  24. Theissen, U. & Martin, W. Sulfide:quinone oxidoreductase (SQR) from the lugworm Arenicola marina shows cyanide- and thioredoxin-dependent activity. FEBS J. 275, 1131–1139 (2008).

    Article  CAS  Google Scholar 

  25. Bella, D.L. & Stipanuk, M.H. Effects of protein, methionine, or chloride on acid-base balance and on cysteine catabolism. Am. J. Physiol. 269, E910–E917 (1995).

    CAS  PubMed  Google Scholar 

  26. Fernandez-Silva, P., Martinez-Azorin, F., Micol, V. & Attardi, G. The human mitochondrial transcription termination factor (mTERF) is a multizipper protein but binds to DNA as a monomer, with evidence pointing to intramolecular leucine zipper interactions. EMBO J. 16, 1066–1079 (1997).

    Article  CAS  Google Scholar 

  27. Tiranti, V. et al. Identification of the gene encoding the human mitochondrial RNA polymerase (h-mtRPOL) by cyberscreening of the Expressed Sequence Tags database. Hum. Mol. Genet. 6, 615–625 (1997).

    Article  CAS  Google Scholar 

  28. Moroni, I., D'Incerti, L., Farina, L., Rimoldi, M. & Uziel, G. Clinical, biochemical and neuroradiological findings in l-2-hydroxyglutaric aciduria. Neurol. Sci. 21, 103–108 (2000).

    Article  CAS  Google Scholar 

  29. Di Donato, S. et al. Systemic carnitine deficiency due to lack of electron transfer flavoprotein:ubiquinone oxidoreductase. Neurology 36, 957–963 (1986).

    Article  CAS  Google Scholar 

  30. Shih, V.E., Carney, M.M. & Mandell, R. A simple screening test for sulfite oxidase deficiency: detection of urinary thiosulfate by a modification of Sorbo's method. Clin. Chim. Acta 95, 143–145 (1979).

    Article  CAS  Google Scholar 

  31. Linden, D.R. et al. Production of the gaseous signal molecule hydrogen sulfide in mouse tissues. J. Neurochem. 106, 1577–1585 (2008).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Furne, J., Saeed, A. & Levitt, M.D. Whole tissue hydrogen sulfide concentrations are orders of magnitude lower than presently accepted values. Am. J. Physiol. Regul. Integr. Comp. Physiol. 295, R1479–R1485 (2008).

    Article  CAS  Google Scholar 

  33. Bennett, M.J. et al. Glutaric aciduria type II: biochemical investigation and treatment of a child diagnosed prenatally. J. Inherit. Metab. Dis. 7, 57–61 (1984).

    Article  CAS  Google Scholar 

  34. Sciacco, M. & Bonilla, E. Cytochemistry and immunocytochemistry of mitochondria in tissue sections. Methods Enzymol. 264, 509–521 (1996).

    Article  CAS  Google Scholar 

  35. Bugiani, M. et al. Clinical and molecular findings in children with complex I deficiency. Biochim. Biophys. Acta 1659, 136–147 (2004).

    Article  CAS  Google Scholar 

  36. Sottocasa, G.L., Kuylenstierna, B., Ernster, L. & Bergstrand, A. An electron-transport system associated with the outer membrane of liver mitochondria. A biochemical and morphological study. J. Cell Biol. 32, 415–438 (1967).

    Article  CAS  Google Scholar 

  37. Sörbo, B. Sulfate: turbidimetric and nephelometric methods. Methods Enzymol. 143, 3–6 (1987).

    Article  Google Scholar 

  38. Sörbo, B. Rhodanese. Methods Enzymol. 2, 334–337 (1955).

    Article  Google Scholar 

  39. Dell'Agnello, C. et al. Increased longevity and refractoriness to Ca2+-dependent neurodegeneration in Surf1 knockout mice. Hum. Mol. Genet. 16, 431–444 (2007).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank A. Bradley (The Wellcome Trust Sanger Institute) for AB1 mouse embryonic stem cells. We are grateful to M. Bada for skillful technical assistance; to B. Garavaglia, E. Lamantea, F. Forlani and M.K. Grieshaber for valuable discussion; to the Chemical Analysis Laboratory, University of Georgia, for metal analysis; and to Primm for MALDI TOF mass spectometry analysis. This work was supported by the Pierfranco and Luisa Mariani Foundation Italy, Fondazione Telethon-Italy grant number GGP07019, the Italian Ministry of University and Research (FIRB 2003—project RBLA038RMA), MITOCIRCLE and EUMITOCOMBAT network grants from the European Union framework program 6 and by the Deutsche Forschungsgemeinschaft (GR 456/22-1).

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Authors and Affiliations

  1. Pierfranco and Luisa Mariani Center for Research on Children's Mitochondrial Disorders, Unit of Molecular Neurogenetics, Institute of Neurology 'Carlo Besta'—Istituto di Ricovero e Cura a Carattere Scientifico (IRCCS) Foundation, Via Temolo 4, Milan, 20126, Italy

    Valeria Tiranti, Carlo Viscomi, Ivano Di Meo, Rossana Mineri & Massimo Zeviani

  2. Biochemistry Institute, Heinrich-Heine-Universität, Düsseldorf, 40225, Germany

    Tatjana Hildebrandt

  3. Foundation European Brain Research Institute (EBRI)—Rita Levi-Montalcini, Disease Modelling Facility, Via del Fosso di Fiorano 64/65, Rome, 00143, Italy

    Cecilia Tiveron

  4. Research Service, Veterans Affairs Medical Center, 1 Veterans Drive, Minneapolis, 55417, Minnesota, USA

    Michael D Levitt

  5. Division of Neurology, Dino Ferrari Center, Ospedale Maggiore Policlinico, Mangiagalli e Regina Elena-IRCCS Foundation, Via F. Sforza 35, Milan, 20122, Italy

    Alessandro Prelle & Gigliola Fagiolari

  6. Unit of Biochemistry and Genetics, Institute of Neurology 'Carlo Besta'-IRCCS Foundation, Via Celoria 11, Milan, 20133, Italy

    Marco Rimoldi

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  1. Valeria Tiranti
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Contributions

V.T. and M.Z. designed the experimental plan and wrote the manuscript; C.V. and C.T. took care of the creation of the Ethe1-knockout mouse; C.V. characterized the mouse phenotype; I.DM. carried out the measurements of the mitochondrial respiratory chain biochemistry; R.M. performed the experiments on expression and purification of the recombinant ETHE1 protein in E. coli; T.H. performed the sulfur-related enzymology; M.D.L. measured the sulfur compounds in body fluids and tissues; G.F. and A.P. carried out the morphological investigation; and M.R. took care of the metabolite analysis in urine and blood.

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Correspondence to Valeria Tiranti or Massimo Zeviani.

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Tiranti, V., Viscomi, C., Hildebrandt, T. et al. Loss of ETHE1, a mitochondrial dioxygenase, causes fatal sulfide toxicity in ethylmalonic encephalopathy. Nat Med 15, 200–205 (2009). https://doi.org/10.1038/nm.1907

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