Pathophysiology of the MELAS 3243 transition mutation

J Biol Chem. 1997 Oct 24;272(43):27189-96. doi: 10.1074/jbc.272.43.27189.


Single base substitutions of the mitochondrial genome are associated with a variety of metabolic disorders. The myopathy, encephalopathy, lactic acidosis, stroke-like episodes syndrome, most frequently associated with an A to G transition mutation at position 3243 of the mitochondrial tRNALeu(UUR) gene, is characterized by biochemical and structural alterations of mitochondria. To investigate the pathophysiology of the mutation, we established distinct Epstein-Barr virus-transformed B-cell lines for analyses that harbored 30-70% of the mutated genome. Interestingly, neither an alteration of the processing of primary transcripts nor a general impairment of individual mitochondrial protein subunit synthesis rates could be observed. Nevertheless a marked decrease of cytochrome-c oxidase activity and reduced content of mitochondrial encoded subunits in the assembled respiratory complex IV was recorded on the cell line harboring 70% mutated mtDNA. Quantitative analysis of incorporation rates of the amino acid leucine into newly synthesized mitochondrial proteins, representing the functionality of the tRNALeu(UUR) in protein biosynthesis, revealed a specific decrease of this amino acid in distinct mitochondrial translation products. This observation was supported by a variation in the proteolytic fingerprint pattern. Our results suggest that the malfunctioning mitochondrial tRNALeu(UUR) leads to an alteration of amino acid incorporation into the mitochondrially synthesized subunits of the oxidative phosphorylation system, thus altering it's structure and function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • B-Lymphocytes
  • Base Sequence
  • Cell Line, Transformed
  • Citrate (si)-Synthase / metabolism
  • DNA Primers
  • DNA, Mitochondrial / genetics*
  • Electron Transport Complex IV / genetics
  • Electron Transport Complex IV / metabolism
  • Herpesvirus 4, Human
  • Humans
  • Kinetics
  • MELAS Syndrome / enzymology
  • MELAS Syndrome / genetics*
  • MELAS Syndrome / physiopathology*
  • Macromolecular Substances
  • Mitochondria / enzymology*
  • Mutation*
  • NADH Dehydrogenase / metabolism
  • Oxidative Phosphorylation
  • Polymerase Chain Reaction
  • Protein Biosynthesis
  • RNA, Transfer, Leu / genetics
  • Reference Values
  • Succinate Cytochrome c Oxidoreductase / metabolism
  • Succinate Dehydrogenase / metabolism


  • DNA Primers
  • DNA, Mitochondrial
  • Macromolecular Substances
  • RNA, Transfer, Leu
  • Succinate Cytochrome c Oxidoreductase
  • Succinate Dehydrogenase
  • NADH Dehydrogenase
  • Electron Transport Complex IV
  • Citrate (si)-Synthase