Complementation of mutant and wild-type human mitochondrial DNAs coexisting since the mutation event and lack of complementation of DNAs introduced separately into a cell within distinct organelles (original) (raw)

Abstract

The rules that govern complementation of mutant and wild-type mitochondrial genomes in human cells were investigated under different experimental conditions. Among mitochondrial transformants derived from an individual affected by the MERRF (myoclonus epilepsy associated with ragged red fibers) encephalomyopathy and carrying in heteroplasmic form the mitochondrial tRNA(Lys) mutation associated with that syndrome, normal protein synthesis and respiration was observed when the wild-type mitochondrial DNA exceeded 10% of the total complement. In these transformants, the protective effect of wild-type mitochondrial DNA was shown to involve interactions of the mutant and wild-type gene products. Very different results were obtained in experiments in which two mitochondrial DNAs carrying nonallelic disease-causing mutations were sequentially introduced within distinct organelles into the same human mitochondrial DNA-less (rho 0) cell. In transformants exhibiting different ratios of the two genomes, no evidence of cooperation between their products was observed, even 3 months after the introduction of the second mutation. These results pointed to the phenotypic independence of the two genomes. A similar conclusion was reached in experiments in which mitochondria carrying a chloramphenicol resistance-inducing mitochondrial DNA mutation were introduced into chloramphenicol-sensitive cells. A plausible interpretation of the different results obtained in the latter two sets of experiments, compared with the complementation behavior observed in the heteroplasmic MERRF transformants, is that in the latter, the mutant and wild-type genomes coexisted in the same organelles from the time of the mutation. This would imply that the way in which mitochondrial DNA is sorted among different organelles plays a fundamental role in determining the oxidative-phosphorylation phenotype in mammalian cells. These results have significant implications for mitochondrial genetics and for studies on the transmission and therapy of mitochondrial DNA-linked diseases.

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  1. Adoutte A., Sainsard A., Rossignol M., Beisson J. Aspects génétiques de la biogenèse des mitochondries chez la Paramécie. Biochimie. 1973;55(6):793–799. [PubMed] [Google Scholar]
  2. Ashley M. V., Laipis P. J., Hauswirth W. W. Rapid segregation of heteroplasmic bovine mitochondria. Nucleic Acids Res. 1989 Sep 25;17(18):7325–7331. doi: 10.1093/nar/17.18.7325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Azpiroz R., Butow R. A. Patterns of mitochondrial sorting in yeast zygotes. Mol Biol Cell. 1993 Jan;4(1):21–36. doi: 10.1091/mbc.4.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Birky C. W., Jr On the origin of mitochondrial mutants: evidence for intracellular selection of mitochondria in the origin of antibiotic-resistant cells in yeast. Genetics. 1973 Jul;74(3):421–432. doi: 10.1093/genetics/74.3.421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Boulet L., Karpati G., Shoubridge E. A. Distribution and threshold expression of the tRNA(Lys) mutation in skeletal muscle of patients with myoclonic epilepsy and ragged-red fibers (MERRF). Am J Hum Genet. 1992 Dec;51(6):1187–1200. [PMC free article] [PubMed] [Google Scholar]
  6. Boynton J. E., Harris E. H., Burkhart B. D., Lamerson P. M., Gillham N. W. Transmission of mitochondrial and chloroplast genomes in crosses of Chlamydomonas. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2391–2395. doi: 10.1073/pnas.84.8.2391. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bunn C. L., Mitchell C. H., Lukins H. B., Linnane A. W. Biogenesis of mitochondria. 18. A new class of cytoplasmically determined antibiotic resistant mutants in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1233–1240. doi: 10.1073/pnas.67.3.1233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chomyn A., Martinuzzi A., Yoneda M., Daga A., Hurko O., Johns D., Lai S. T., Nonaka I., Angelini C., Attardi G. MELAS mutation in mtDNA binding site for transcription termination factor causes defects in protein synthesis and in respiration but no change in levels of upstream and downstream mature transcripts. Proc Natl Acad Sci U S A. 1992 May 15;89(10):4221–4225. doi: 10.1073/pnas.89.10.4221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chomyn A., Meola G., Bresolin N., Lai S. T., Scarlato G., Attardi G. In vitro genetic transfer of protein synthesis and respiration defects to mitochondrial DNA-less cells with myopathy-patient mitochondria. Mol Cell Biol. 1991 Apr;11(4):2236–2244. doi: 10.1128/mcb.11.4.2236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clark-Walker G. D., Miklos G. L. Complementation in cytoplasmic petite mutants of yeast to form respiratory competent cells. Proc Natl Acad Sci U S A. 1975 Jan;72(1):372–375. doi: 10.1073/pnas.72.1.372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Clayton D. A. Replication of animal mitochondrial DNA. Cell. 1982 Apr;28(4):693–705. doi: 10.1016/0092-8674(82)90049-6. [DOI] [PubMed] [Google Scholar]
  12. Corral-Debrinski M., Horton T., Lott M. T., Shoffner J. M., Beal M. F., Wallace D. C. Mitochondrial DNA deletions in human brain: regional variability and increase with advanced age. Nat Genet. 1992 Dec;2(4):324–329. doi: 10.1038/ng1292-324. [DOI] [PubMed] [Google Scholar]
  13. Cortopassi G. A., Arnheim N. Detection of a specific mitochondrial DNA deletion in tissues of older humans. Nucleic Acids Res. 1990 Dec 11;18(23):6927–6933. doi: 10.1093/nar/18.23.6927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Cortopassi G. A., Shibata D., Soong N. W., Arnheim N. A pattern of accumulation of a somatic deletion of mitochondrial DNA in aging human tissues. Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7370–7374. doi: 10.1073/pnas.89.16.7370. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Gonzalez I. L., Gorski J. L., Campen T. J., Dorney D. J., Erickson J. M., Sylvester J. E., Schmickel R. D. Variation among human 28S ribosomal RNA genes. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7666–7670. doi: 10.1073/pnas.82.22.7666. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Goto Y., Nonaka I., Horai S. A mutation in the tRNA(Leu)(UUR) gene associated with the MELAS subgroup of mitochondrial encephalomyopathies. Nature. 1990 Dec 13;348(6302):651–653. doi: 10.1038/348651a0. [DOI] [PubMed] [Google Scholar]
  18. Gurgo C., Apirion D., Schlessinger D. Polyribosome metabolism in Escherichia coli treated with chloramphenicol, neomycin, spectinomycin or tetracycline. J Mol Biol. 1969 Oct 28;45(2):205–220. doi: 10.1016/0022-2836(69)90100-4. [DOI] [PubMed] [Google Scholar]
  19. Hayakawa M., Hattori K., Sugiyama S., Ozawa T. Age-associated oxygen damage and mutations in mitochondrial DNA in human hearts. Biochem Biophys Res Commun. 1992 Dec 15;189(2):979–985. doi: 10.1016/0006-291x(92)92300-m. [DOI] [PubMed] [Google Scholar]
  20. Hayashi J., Ohta S., Kikuchi A., Takemitsu M., Goto Y., Nonaka I. Introduction of disease-related mitochondrial DNA deletions into HeLa cells lacking mitochondrial DNA results in mitochondrial dysfunction. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10614–10618. doi: 10.1073/pnas.88.23.10614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hayashi J., Werbin H., Shay J. W. Effects of normal human fibroblast mitochondrial DNA on segregation of HeLaTG Mitochondrial DNA and on tumorigenicity of HeLaTG cells. Cancer Res. 1986 Aug;46(8):4001–4006. [PubMed] [Google Scholar]
  22. Howell N., Halvorson S., Kubacka I., McCullough D. A., Bindoff L. A., Turnbull D. M. Mitochondrial gene segregation in mammals: is the bottleneck always narrow? Hum Genet. 1992 Sep-Oct;90(1-2):117–120. doi: 10.1007/BF00210753. [DOI] [PubMed] [Google Scholar]
  23. King M. P., Attardi G. Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. Science. 1989 Oct 27;246(4929):500–503. doi: 10.1126/science.2814477. [DOI] [PubMed] [Google Scholar]
  24. King M. P., Attardi G. Injection of mitochondria into human cells leads to a rapid replacement of the endogenous mitochondrial DNA. Cell. 1988 Mar 25;52(6):811–819. doi: 10.1016/0092-8674(88)90423-0. [DOI] [PubMed] [Google Scholar]
  25. Kobayashi Y., Momoi M. Y., Tominaga K., Momoi T., Nihei K., Yanagisawa M., Kagawa Y., Ohta S. A point mutation in the mitochondrial tRNA(Leu)(UUR) gene in MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes). Biochem Biophys Res Commun. 1990 Dec 31;173(3):816–822. doi: 10.1016/s0006-291x(05)80860-5. [DOI] [PubMed] [Google Scholar]
  26. Koehler C. M., Lindberg G. L., Brown D. R., Beitz D. C., Freeman A. E., Mayfield J. E., Myers A. M. Replacement of bovine mitochondrial DNA by a sequence variant within one generation. Genetics. 1991 Sep;129(1):247–255. doi: 10.1093/genetics/129.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Mita S., Schmidt B., Schon E. A., DiMauro S., Bonilla E. Detection of "deleted" mitochondrial genomes in cytochrome-c oxidase-deficient muscle fibers of a patient with Kearns-Sayre syndrome. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9509–9513. doi: 10.1073/pnas.86.23.9509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Mitchell C. H., England J. M., Attardi G. Isolation of chloramphenicol-resistant variants from a human cell line. Somatic Cell Genet. 1975 Jul;1(3):215–234. doi: 10.1007/BF01538447. [DOI] [PubMed] [Google Scholar]
  31. Mounkes L. C., Jones R. S., Liang B. C., Gelbart W., Fuller M. T. A Drosophila model for xeroderma pigmentosum and Cockayne's syndrome: haywire encodes the fly homolog of ERCC3, a human excision repair gene. Cell. 1992 Dec 11;71(6):925–937. doi: 10.1016/0092-8674(92)90389-t. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Oliver N. A., Wallace D. C. Assignment of two mitochondrially synthesized polypeptides to human mitochondrial DNA and their use in the study of intracellular mitochondrial interaction. Mol Cell Biol. 1982 Jan;2(1):30–41. doi: 10.1128/mcb.2.1.30. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Olivo P. D., Van de Walle M. J., Laipis P. J., Hauswirth W. W. Nucleotide sequence evidence for rapid genotypic shifts in the bovine mitochondrial DNA D-loop. Nature. 1983 Nov 24;306(5941):400–402. doi: 10.1038/306400a0. [DOI] [PubMed] [Google Scholar]
  35. 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]
  36. 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]
  37. Shoubridge E. A., Karpati G., Hastings K. E. Deletion mutants are functionally dominant over wild-type mitochondrial genomes in skeletal muscle fiber segments in mitochondrial disease. Cell. 1990 Jul 13;62(1):43–49. doi: 10.1016/0092-8674(90)90238-a. [DOI] [PubMed] [Google Scholar]
  38. Smiley S. T., Reers M., Mottola-Hartshorn C., Lin M., Chen A., Smith T. W., Steele G. D., Jr, Chen L. B. Intracellular heterogeneity in mitochondrial membrane potentials revealed by a J-aggregate-forming lipophilic cation JC-1. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3671–3675. doi: 10.1073/pnas.88.9.3671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Soong N. W., Hinton D. R., Cortopassi G., Arnheim N. Mosaicism for a specific somatic mitochondrial DNA mutation in adult human brain. Nat Genet. 1992 Dec;2(4):318–323. doi: 10.1038/ng1292-318. [DOI] [PubMed] [Google Scholar]
  40. Spolsky C. M., Eisenstadt J. M. Chloramphenicol-resistant mutants of human HeLa cells. FEBS Lett. 1972 Sep 15;25(2):319–324. doi: 10.1016/0014-5793(72)80514-3. [DOI] [PubMed] [Google Scholar]
  41. Tanaka M., Ino H., Ohno K., Ohbayashi T., Ikebe S., Sano T., Ichiki T., Kobayashi M., Wada Y., Ozawa T. Mitochondrial DNA mutations in mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Biochem Biophys Res Commun. 1991 Jan 31;174(2):861–868. doi: 10.1016/0006-291x(91)91497-z. [DOI] [PubMed] [Google Scholar]
  42. Wallace D. C. Assignment of the chloramphenicol resistance gene to mitochondrial deoxyribonucleic acid and analysis of its expression in cultured human cells. Mol Cell Biol. 1981 Aug;1(8):697–710. doi: 10.1128/mcb.1.8.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wallace D. C. Diseases of the mitochondrial DNA. Annu Rev Biochem. 1992;61:1175–1212. doi: 10.1146/annurev.bi.61.070192.005523. [DOI] [PubMed] [Google Scholar]
  44. White F. A., Bunn C. L. Segregation of mitochondrial DNA in human somatic cell hybrids. Mol Gen Genet. 1984;197(3):453–460. doi: 10.1007/BF00329942. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. Yoneda M., Chomyn A., Martinuzzi A., Hurko O., Attardi G. Marked replicative advantage of human mtDNA carrying a point mutation that causes the MELAS encephalomyopathy. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11164–11168. doi: 10.1073/pnas.89.23.11164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yoneda M., Tanno Y., Horai S., Ozawa T., Miyatake T., Tsuji S. A common mitochondrial DNA mutation in the t-RNA(Lys) of patients with myoclonus epilepsy associated with ragged-red fibers. Biochem Int. 1990 Aug;21(5):789–796. [PubMed] [Google Scholar]
  48. Yoneda M., Tanno Y., Nonaka I., Miyatake T., Tsuji S. Simple detection of tRNA(Lys) mutation in myoclonus epilepsy associated with ragged-red fibers (MERRF) by polymerase chain reaction with a mismatched primer. Neurology. 1991 Nov;41(11):1838–1840. doi: 10.1212/wnl.41.11.1838. [DOI] [PubMed] [Google Scholar]