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

Kenta Akama, Brooke E. Christian, Christie N. Jones, Takuya Ueda, Nono Takeuchi, Linda L. Spremulli

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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-Tsmt) and elongation factor Tu (EF-Tumt). EF-Tumt delivers the aminoacyl-tRNA (aa-tRNA) to the ribosome during the elongation phase of protein synthesis. EF-Tsmt regenerates EF-Tumt:GTP from EF-Tumt:GDP. A mutation of EF-Tsmt (R325W) leads to a two-fold reduction in its ability to stimulate the activity of EF-Tumt in poly(U)-directed polypeptide chain elongation. This loss of activity is caused by a significant reduction in the ability of EF-Tsmt R325W to bind EF-Tumt, leading to a defect in nucleotide exchange. A mutation of Arg336 to Gln in EF-Tumt causes infantile encephalopathy caused by defects in mitochondrial translation. EF-Tumt R336Q is as active as the wild-type protein in polymerization using Escherichia coli 70S ribosomes and E. coli [14C]Phe-tRNA but is inactive in polymerization with mitochondrial [14C]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-Tumt for binding to mitochondrial aa-tRNA. Clearly the R336Q mutation in EF-Tumt has a far more drastic effect on its interaction with mitochondrial aa-tRNAs than bacterial aa-tRNAs.

Original languageEnglish
Pages (from-to)692-698
Number of pages7
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1802
Issue number7-8
DOIs
Publication statusPublished - 2010 Jul 1
Externally publishedYes

Fingerprint

Peptide Elongation Factors
Transfer RNA
Mutation
RNA, Transfer, Phe
Escherichia coli
Ribosomes
Polymerization
Proteins
Peptide Elongation Factor Tu
Poly U
Oxidative Phosphorylation
Brain Diseases
Guanosine Triphosphate
Genes
Mitochondria
Nucleotides
Genome
Peptides

Keywords

  • Disease
  • EF-Ts
  • EF-Tu
  • Elongation
  • Mitochondria
  • Translation

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine

Cite this

Analysis of the functional consequences of lethal mutations in mitochondrial translational elongation factors. / Akama, Kenta; Christian, Brooke E.; Jones, Christie N.; Ueda, Takuya; Takeuchi, Nono; Spremulli, Linda L.

In: Biochimica et Biophysica Acta - Molecular Basis of Disease, Vol. 1802, No. 7-8, 01.07.2010, p. 692-698.

Research output: Contribution to journalArticle

Akama, Kenta ; Christian, Brooke E. ; Jones, Christie N. ; Ueda, Takuya ; Takeuchi, Nono ; Spremulli, Linda L. / Analysis of the functional consequences of lethal mutations in mitochondrial translational elongation factors. In: Biochimica et Biophysica Acta - Molecular Basis of Disease. 2010 ; Vol. 1802, No. 7-8. pp. 692-698.
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