Glutamyl-tRNAGln amidotransferase is essential for mammalian mitochondrial translation in vivo

Biochem J. 2014 May 15;460(1):91-101. doi: 10.1042/BJ20131107.

Abstract

Translational accuracy depends on the correct formation of aminoacyl-tRNAs, which, in the majority of cases, are produced by specific aminoacyl-tRNA synthetases that ligate each amino acid to its cognate isoaceptor tRNA. Aminoacylation of tRNAGln, however, is performed by various mechanisms in different systems. Since no mitochondrial glutaminyl-tRNA synthetase has been identified to date in mammalian mitochondria, Gln-tRNAGln has to be formed by an indirect mechanism in the organelle. It has been demonstrated that human mitochondria contain a non-discriminating glutamyl-tRNA synthetase and the heterotrimeric enzyme GatCAB (where Gat is glutamyl-tRNAGln amidotransferase), which are able to catalyse the formation of Gln-tRNAGln in vitro. In the present paper we demonstrate that mgatA (mouse GatA) interference in mouse cells produces a strong defect in mitochondrial translation without affecting the stability of the newly synthesized proteins. As a result, interfered cells present an impairment of the oxidative phosphorylation system and a significant increase in ROS (reactive oxygen species) levels. MS analysis of mitochondrial proteins revealed no glutamic acid found in the position of glutamines, strongly suggesting that misaminoacylated Glu-tRNAGln is rejected from the translational apparatus to maintain the fidelity of mitochondrial protein synthesis in mammals.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Mice
  • Mitochondria / enzymology*
  • Mitochondria / genetics*
  • Nitrogenous Group Transferases / genetics*
  • Nitrogenous Group Transferases / metabolism*
  • Oxidative Phosphorylation
  • Phenotype
  • Protein Biosynthesis / physiology*
  • Protein Stability
  • Reactive Oxygen Species / metabolism
  • Tandem Mass Spectrometry

Substances

  • Reactive Oxygen Species
  • Nitrogenous Group Transferases
  • glutamyl-tRNA(Gln) amidotransferase