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. 1991 Apr;11(4):2236–2244. doi: 10.1128/mcb.11.4.2236

In vitro genetic transfer of protein synthesis and respiration defects to mitochondrial DNA-less cells with myopathy-patient mitochondria.

A Chomyn 1, G Meola 1, N Bresolin 1, S T Lai 1, G Scarlato 1, G Attardi 1
PMCID: PMC359920  PMID: 1848674

Abstract

A severe mitochondrial protein synthesis defect in myoblasts from a patient with mitochondrial myopathy was transferred with myoblast mitochondria into two genetically unrelated mitochondrial DNA (mtDNA)-less human cell lines, pointing to an mtDNA alteration as being responsible and sufficient for causing the disease. The transfer of the defect correlated with marked deficiencies in respiration and cytochrome c oxidase activity of the transformants and the presence in their mitochondria of mtDNA carrying a tRNA(Lys) mutation. Furthermore, apparently complete segregation of the defective genotype and phenotype was observed in the transformants derived from the heterogeneous proband myoblast population, suggesting that the mtDNA heteroplasmy in this population was to a large extent intercellular. The present work thus establishes a direct link between mtDNA alteration and a biochemical defect.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson S., Bankier A. T., Barrell B. G., de Bruijn M. H., Coulson A. R., Drouin J., Eperon I. C., Nierlich D. P., Roe B. A., Sanger F. Sequence and organization of the human mitochondrial genome. Nature. 1981 Apr 9;290(5806):457–465. doi: 10.1038/290457a0. [DOI] [PubMed] [Google Scholar]
  2. Attardi G., Schatz G. Biogenesis of mitochondria. Annu Rev Cell Biol. 1988;4:289–333. doi: 10.1146/annurev.cb.04.110188.001445. [DOI] [PubMed] [Google Scholar]
  3. Capaldi R. A. Mitochondrial myopathies and respiratory chain proteins. Trends Biochem Sci. 1988 Apr;13(4):144–148. doi: 10.1016/0968-0004(88)90073-4. [DOI] [PubMed] [Google Scholar]
  4. Ching E., Attardi G. High-resolution electrophoretic fractionation and partial characterization of the mitochondrial translation products from HeLa cells. Biochemistry. 1982 Jun 22;21(13):3188–3195. doi: 10.1021/bi00256a024. [DOI] [PubMed] [Google Scholar]
  5. DiMauro S., Bonilla E., Zeviani M., Nakagawa M., DeVivo D. C. Mitochondrial myopathies. Ann Neurol. 1985 Jun;17(6):521–538. doi: 10.1002/ana.410170602. [DOI] [PubMed] [Google Scholar]
  6. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  7. Fung C. H., Khachadurian A. K. Suppression of synthesis and esterification of cholesterol and stimulation of low density lipoprotein receptor activity by polyoxyethylated cholesterol in cultured human fibroblasts. J Biol Chem. 1980 Jan 25;255(2):676–680. [PubMed] [Google Scholar]
  8. Holt I. J., Harding A. E., Morgan-Hughes J. A. Deletions of muscle mitochondrial DNA in patients with mitochondrial myopathies. Nature. 1988 Feb 25;331(6158):717–719. doi: 10.1038/331717a0. [DOI] [PubMed] [Google Scholar]
  9. Holt I. J., Harding A. E., Petty R. K., Morgan-Hughes J. A. A new mitochondrial disease associated with mitochondrial DNA heteroplasmy. Am J Hum Genet. 1990 Mar;46(3):428–433. [PMC free article] [PubMed] [Google Scholar]
  10. Holt I. J., Miller D. H., Harding A. E. Genetic heterogeneity and mitochondrial DNA heteroplasmy in Leber's hereditary optic neuropathy. J Med Genet. 1989 Dec;26(12):739–743. doi: 10.1136/jmg.26.12.739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kim S. H., Suddath F. L., Quigley G. J., McPherson A., Sussman J. L., Wang A. H., Seeman N. C., Rich A. Three-dimensional tertiary structure of yeast phenylalanine transfer RNA. Science. 1974 Aug 2;185(4149):435–440. doi: 10.1126/science.185.4149.435. [DOI] [PubMed] [Google Scholar]
  12. Meola G., Scarpini E., Velicogna M., Mottura A., Baron P. L., Beretta S., Scarlato G. Analysis of fibronectin expression during human muscle differentiation. Basic Appl Histochem. 1986;30(2):153–163. [PubMed] [Google Scholar]
  13. Mitchell C. H., Attardi G. Cytoplasmic transfer of chloramphenicol resistance in a human cell line. Somatic Cell Genet. 1978 Nov;4(6):737–744. doi: 10.1007/BF01543161. [DOI] [PubMed] [Google Scholar]
  14. Moraes C. T., Schon E. A., DiMauro S., Miranda A. F. Heteroplasmy of mitochondrial genomes in clonal cultures from patients with Kearns-Sayre syndrome. Biochem Biophys Res Commun. 1989 Apr 28;160(2):765–771. doi: 10.1016/0006-291x(89)92499-6. [DOI] [PubMed] [Google Scholar]
  15. Normanly J., Abelson J. tRNA identity. Annu Rev Biochem. 1989;58:1029–1049. doi: 10.1146/annurev.bi.58.070189.005121. [DOI] [PubMed] [Google Scholar]
  16. Oliver N. A., Greenberg B. D., Wallace D. C. Assignment of a polymorphic polypeptide to the human mitochondrial DNA unidentified reading frame 3 gene by a new peptide mapping strategy. J Biol Chem. 1983 May 10;258(9):5834–5839. [PubMed] [Google Scholar]
  17. Ozawa T., Yoneda M., Tanaka M., Ohno K., Sato W., Suzuki H., Nishikimi M., Yamamoto M., Nonaka I., Horai S. Maternal inheritance of deleted mitochondrial DNA in a family with mitochondrial myopathy. Biochem Biophys Res Commun. 1988 Aug 15;154(3):1240–1247. doi: 10.1016/0006-291x(88)90272-0. [DOI] [PubMed] [Google Scholar]
  18. Rich A., RajBhandary U. L. Transfer RNA: molecular structure, sequence, and properties. Annu Rev Biochem. 1976;45:805–860. doi: 10.1146/annurev.bi.45.070176.004105. [DOI] [PubMed] [Google Scholar]
  19. Rosing H. S., Hopkins L. C., Wallace D. C., Epstein C. M., Weidenheim K. Maternally inherited mitochondrial myopathy and myoclonic epilepsy. Ann Neurol. 1985 Mar;17(3):228–237. doi: 10.1002/ana.410170303. [DOI] [PubMed] [Google Scholar]
  20. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Shoffner J. M., Lott M. T., Lezza A. M., Seibel P., Ballinger S. W., Wallace D. C. Myoclonic epilepsy and ragged-red fiber disease (MERRF) is associated with a mitochondrial DNA tRNA(Lys) mutation. Cell. 1990 Jun 15;61(6):931–937. doi: 10.1016/0092-8674(90)90059-n. [DOI] [PubMed] [Google Scholar]
  22. Storrie B., Attardi G. Expression of the mitochondrial genome in HeLa cells. 13. Effect of selective inhibition of cytoplasmic or mitochondrial protein synthesis on mitochondrial nucleic acid synthesis. J Mol Biol. 1972 Nov 14;71(2):177–199. doi: 10.1016/0022-2836(72)90345-2. [DOI] [PubMed] [Google Scholar]
  23. Vilkki J., Savontaus M. L., Nikoskelainen E. K. Genetic heterogeneity in Leber hereditary optic neuroretinopathy revealed by mitochondrial DNA polymorphism. Am J Hum Genet. 1989 Aug;45(2):206–211. [PMC free article] [PubMed] [Google Scholar]
  24. Wallace D. C. Mitochondrial DNA mutations and neuromuscular disease. Trends Genet. 1989 Jan;5(1):9–13. doi: 10.1016/0168-9525(89)90005-x. [DOI] [PubMed] [Google Scholar]
  25. Wallace D. C. Mitotic segregation of mitochondrial DNAs in human cell hybrids and expression of chloramphenicol resistance. Somat Cell Mol Genet. 1986 Jan;12(1):41–49. doi: 10.1007/BF01560726. [DOI] [PubMed] [Google Scholar]
  26. Wallace D. C., Singh G., Lott M. T., Hodge J. A., Schurr T. G., Lezza A. M., Elsas L. J., 2nd, Nikoskelainen E. K. Mitochondrial DNA mutation associated with Leber's hereditary optic neuropathy. Science. 1988 Dec 9;242(4884):1427–1430. doi: 10.1126/science.3201231. [DOI] [PubMed] [Google Scholar]
  27. Wiseman A., Attardi G. Cytoplasmically determined human cell mutants defective in mitochondrial ribosome assembly. Mol Gen Genet. 1982;186(3):364–371. doi: 10.1007/BF00729455. [DOI] [PubMed] [Google Scholar]
  28. Zeviani M., Moraes C. T., DiMauro S., Nakase H., Bonilla E., Schon E. A., Rowland L. P. Deletions of mitochondrial DNA in Kearns-Sayre syndrome. Neurology. 1988 Sep;38(9):1339–1346. doi: 10.1212/wnl.38.9.1339. [DOI] [PubMed] [Google Scholar]

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