Analysis of the functional consequences of lethal mutations in mitochondrial translational elongation factors

Biochim Biophys Acta. Jul-Aug 2010;1802(7-8):692-8. doi: 10.1016/j.bbadis.2010.04.003. Epub 2010 May 6.


Mammalian mitochondria synthesize a set of thirteen proteins that are essential for energy generation via oxidative phosphorylation. The genes for all of the factors required for synthesis of the mitochondrially encoded proteins are located in the nuclear genome. A number of disease-causing mutations have been identified in these genes. In this manuscript, we have elucidated the mechanisms of translational failure for two disease states characterized by lethal mutations in mitochondrial elongation factor Ts (EF-Ts(mt)) and elongation factor Tu (EF-Tu(mt)). EF-Tu(mt) delivers the aminoacyl-tRNA (aa-tRNA) to the ribosome during the elongation phase of protein synthesis. EF-Ts(mt) regenerates EF-Tu(mt):GTP from EF-Tu(mt):GDP. A mutation of EF-Ts(mt) (R325W) leads to a two-fold reduction in its ability to stimulate the activity of EF-Tu(mt) in poly(U)-directed polypeptide chain elongation. This loss of activity is caused by a significant reduction in the ability of EF-Ts(mt) R325W to bind EF-Tu(mt), leading to a defect in nucleotide exchange. A mutation of Arg336 to Gln in EF-Tu(mt) causes infantile encephalopathy caused by defects in mitochondrial translation. EF-Tu(mt) R336Q is as active as the wild-type protein in polymerization using Escherichia coli 70S ribosomes and E. coli [(14)C]Phe-tRNA but is inactive in polymerization with mitochondrial [(14)C]Phe-tRNA and mitochondrial 55S ribosomes. The R336Q mutation causes a two-fold decrease in ternary complex formation with E. coli aa-tRNA but completely inactivates EF-Tu(mt) for binding to mitochondrial aa-tRNA. Clearly the R336Q mutation in EF-Tu(mt) has a far more drastic effect on its interaction with mitochondrial aa-tRNAs than bacterial aa-tRNAs.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution / physiology
  • Animals
  • Cattle
  • Escherichia coli Proteins / genetics
  • Escherichia coli Proteins / metabolism
  • Escherichia coli Proteins / physiology
  • Genes, Lethal* / physiology
  • Mitochondria / genetics
  • Mitochondria / metabolism*
  • Models, Molecular
  • Mutant Proteins / chemistry
  • Mutant Proteins / metabolism
  • Mutant Proteins / physiology
  • Mutation* / physiology
  • Peptide Elongation Factor Tu / chemistry
  • Peptide Elongation Factor Tu / genetics
  • Peptide Elongation Factor Tu / metabolism
  • Peptide Elongation Factor Tu / physiology
  • Peptide Elongation Factors / analysis
  • Peptide Elongation Factors / chemistry
  • Peptide Elongation Factors / genetics*
  • Peptide Elongation Factors / metabolism
  • Peptide Elongation Factors / physiology*
  • Protein Binding
  • Protein Biosynthesis / genetics*
  • Protein Multimerization
  • RNA, Transfer, Amino Acid-Specific / metabolism
  • Structure-Activity Relationship


  • Escherichia coli Proteins
  • Mutant Proteins
  • Peptide Elongation Factors
  • RNA, Transfer, Amino Acid-Specific
  • elongation factor Ts
  • Peptide Elongation Factor Tu