We have investigated the pathogenetic mechanism of the mitochondrial tRNALys gene mutation (position 8344) associated with MERRF encephalomyopathy in several mitochondrial DMA (mtDNA)–less cell transformants carrying the mutation and in control cells. A decrease of 50–60% in the specific tRNALys aminoacylation capacity per cell was found in mutant cells. Furthermore, several lines of evidence reveal that the severe protein synthesis impairment in MERRF mutation–carrying cells is due to premature termination of translation at each or near each lysine codon, with the deficiency of aminoacylated tRNALys being the most likely cause of this phenomenon.
This is a preview of subscription content
Subscribe to Journal
Get full journal access for 1 year
only $4.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Wallace, D.C. Diseases of the mitochondrial DMA. A. Rev. Biochem. 61, 1175–1212 (1992).
Shoffner, J.M. et al. Myoclonic epilepsy and ragged-red fiber disease (MERRF) is associated with a mitochondrial DMA tRNALys mutation. Cell 61, 931–937 (1990).
Chomyn, A. et al. In vitro genetic transfer of protein synthesis and respiration defects to mitochondrial DNA-less cells with myopathy-patient mitochondria. Molec. cell Biol. 11, 2236–2244 (1991).
Boulet, L., Karpati, G. & Shoubridge, E.A. Distribution and threshold expression of the tRNALys mutation in skeletal muscle of patients with myoclonic epilepsy and ragged-red fibers (MERRF). Am. J. hum. Genet. 51, 1187–1200 (1992).
Yoneda, M., Miyatake, T. & Attardi, G. Complementation of mutant and wild-type human mitochondrial DMAs coexisting since the mutation event and lack of complementation of DMAs introduced separately into a cell within distinct organelles. Molec. cell Biol. 14, 2699–2712 (1994).
Wallace, D.C. et al. Familial mitochondrial encephalomyopathy (MERRF): Genetic, pathophysiological, and biochemical characterization of a mitochondrial DMA disease. Cell 55, 601–610 (1988).
King, M.P. & Attardi, G. Human cells lacking mtDNA: repopulation with exogenous mitochondria by complementation. Science 246, 500–503 (1989).
King, M.P. & Attardi, G. Post-transcriptional regulation of the steady-state levels of mitochondrial transfer RNAs in HeLa cells. J. biol. Chem. 268, 10228–10237 (1993).
Ojala, D., Montoya, J., & Attardi, G. tRNA punctuation model of RNA processing in human mitochondria. Nature 290, 470–474 (1981).
Attardi, G., Doersen, C., Gaines, G., King, M. & Montoya, J. New insights into the mechanisms of RNA synthesis and processing in human mitochondria in Achievements and Perspectives in Mitochondrial Research, vol. II (eds E. Quagliarello etal.) 145–163, (Elsevier, Amsterdam, 1985).
Varshney, U., Lee, C.-P & RajBhandary, U.L. Direct analysis of aminoacylation levels of tRNAs in vivo. J. biol. Chem. 266, 24712–24718 (1991).
Attardi, G. Animal mitochondrial DNA: An extreme example of genetic economy. Int. Rev. Cyt. 93, 93–145 (1985).
Anderson, S. et al. Sequence and organization of the human mitochondrial genome. Nature 290, 427–465 (1981).
Cleveland, D.W., Fischer, S.G., Kirschner, M.W. & Laemmli, U.K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J. biol. Chem. 252, 1102–1106 (1977).
Mariottini, P., Chomyn, A., Riley, M., Cottrell, B., Doolittle, R.F. & Attardi, G. Identification of the polypeptides encoded in the unassigned reading frames 2,4,4L and 5 of human mitochondrial DNA. Proc. natn. Acad. Sci. U.S.A. 83, 1563–1567 (1986).
Attardi, G. & Ojala, D. Mitochondrial ribosomes in HeLa cells. Nature new Biol. 229, 133–136 (1971).
Ojala, D. & Attardi, G. Expression of the mitochondrial genome in HeLa cells. X. Properties of mitochondrial polysomes. J. molec. Biol. 65, 273–289 (1972).
Attardi, B., Cravioto, B. & Attardi, G. Membrane-bound ribosomes in HeLa cells. I. Their proportion to total cell ribosomes and their association with messenger RNA. J. molec. Biol. 44, 47–70 (1969).
Nierhaus, K.H., Franceschi, F., Subramanian, A.R., Erdmann, V.A. & Wittmann-Liebold, B. (eds). The translation apparatus: Structure, function, regulation, evolution. (Plenum Press, New York 1993).
Normanly, J. & Abelson, J. tRNA identity. A Rev. Biochem. 58, 1029–1049 (1989).
Schimmel, P., Giegé, R., Moras, D. & Yokoyama, S. An operational RNA code for amino acids and possible relationship to genetic code. Proc. natn. Acad. Sci. U.S.A. 90, 8763–8768 (1993).
Saks, M.E., Sampson, J.R. & Abelson, J.N. The transfer RNA identity problem: a search for rules. Science 263, 191–197 (1994).
Giegé, R., Puglisi, J.D. & Florentz, C. tRNA structure and aminoacylation efficiency. Progr. nucl. Acid Res. Molec. Biol. 45, 129–206 (1993).
Kurland, C.G. Translational accuracy and the fitness of bacteria. A. Rev. Genet. 26, 29–50 (1992).
Kurland, C.G. & Gallant, J.A. The secret life of the ribosome. In “Accuracy in Molecular Processes”(eds Kirkwood, T.B.L, Rosenberger, R.F. & Galas, D. J.), 127–157 (Chapman and Hall, London, 1986).
Hayashi, J.I. et al. Introduction of disease-related mitochondrial DNA deletions into HeLa cells lacking mitochondrial DNA results in mitochondrial dysfunction. Proc. natn. Acad. Sci. U.S.A. 88, 10614–10618 (1991).
Amaldi, F. & Attardi, G. Partial sequence analysis of ribosomal RNA from HeLa cells. I. Oligonucleotide pattern of 28S and 18S RNA after pancreatic ribonuclease digestion. J. molec. Biol. 33, 737–755 (1968).
Gaines, G. & Attardi, G. Highly efficient RNA-synthesizing system that uses isolated human mitochondria: New initiation events and in vivo-like processing patterns. Molec. cell. Biol. 4, 1605–1617 (1984).
Lynch, D.C. & Attardi, G. Amino acid specificity of the transfer RNA species coded for by HeLa cell mitochondrial DNA. J. molec. Biol. 102, 125–141 (1976).
Chomyn, A. et al. 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. natn. Acad. Sci. U.S.A. 89, 4221–4225.
Feinberg, A.P. & Vogelstein, B.A. technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal. Biochem. 132, 6–13 (1983).
About this article
Cite this article
Enriquez, J., Chomyn, A. & Attardi, G. MtDNA mutation in MERRF syndrome causes defective aminoacylation of tRNALys and premature translation termination. Nat Genet 10, 47–55 (1995). https://doi.org/10.1038/ng0595-47
Patient-specific neural progenitor cells derived from induced pluripotent stem cells offer a promise of good models for mitochondrial disease
Cell and Tissue Research (2020)
Impaired Bioenergetics in Mutant Mitochondrial DNA Determines Cell Fate During Seizure-Like Activity
Molecular Neurobiology (2019)
Nature Reviews Molecular Cell Biology (2018)
Nature Communications (2018)
Structural significance of modified nucleoside 5-taurinomethyl-2-thiouridine, τm5s2U, found at ‘wobble’ position in anticodon loop of human mitochondrial tRNALys
Structural Chemistry (2016)