Overexpression of mitochondrial histidyl-tRNA synthetase restores mitochondrial dysfunction caused by a deafness-associated tRNAHis mutation

J Biol Chem. 2020 Jan 24;295(4):940-954. doi: 10.1074/jbc.RA119.010998. Epub 2019 Dec 9.


The deafness-associated m.12201T>C mutation affects the A5-U68 base-pairing within the acceptor stem of mitochondrial tRNAHis The primary defect in this mutation is an alteration in tRNAHis aminoacylation. Here, we further investigate the molecular mechanism of the deafness-associated tRNAHis 12201T>C mutation and test whether the overexpression of the human mitochondrial histidyl-tRNA synthetase gene (HARS2) in cytoplasmic hybrid (cybrid) cells carrying the m.12201T>C mutation reverses mitochondrial dysfunctions. Using molecular dynamics simulations, we demonstrate that the m.12201T>C mutation perturbs the tRNAHis structure and function, supported by decreased melting temperature, conformational changes, and instability of mutated tRNA. We show that the m.12201T>C mutation-induced alteration of aminoacylation tRNAHis causes mitochondrial translational defects and respiratory deficiency. We found that the transfer of HARS2 into the cybrids carrying the m.12201T>C mutation raises the levels of aminoacylated tRNAHis from 56.3 to 75.0% but does not change the aminoacylation of other tRNAs. Strikingly, HARS2 overexpression increased the steady-state levels of tRNAHis and of noncognate tRNAs, including tRNAAla, tRNAGln, tRNAGlu, tRNALeu(UUR), tRNALys, and tRNAMet, in cells bearing the m.12201T>C mutation. This improved tRNA metabolism elevated the efficiency of mitochondrial translation, activities of oxidative phosphorylation complexes, and respiration capacity. Furthermore, HARS2 overexpression markedly increased mitochondrial ATP levels and membrane potential and reduced production of reactive oxygen species in cells carrying the m.12201T>C mutation. These results indicate that HARS2 overexpression corrects the mitochondrial dysfunction caused by the tRNAHis mutation. These findings provide critical insights into the pathophysiology of mitochondrial disease and represent a step toward improved therapeutic interventions for mitochondrial disorders.

Keywords: ATP; aminoacyl tRNA synthetase; aminoacylation; deafness; gene transfer; hearing; m.12201T>C mutation; mitochondrial DNA (mtDNA); mitochondrial histidyl-tRNA synthetase; mitochondrial membrane potential; mitochondrial respiratory chain complex; mitochondrial tRNA mutation; oxidative phosphorylation system (OXPHOS); pathophysiology; reactive oxygen species (ROS); rescue; transfer RNA (tRNA); translation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Amino Acyl-tRNA Synthetases / chemistry
  • Amino Acyl-tRNA Synthetases / genetics*
  • Amino Acyl-tRNA Synthetases / metabolism
  • Aminoacylation
  • Cell Line
  • Cell Respiration
  • DNA, Mitochondrial / metabolism
  • Deafness / enzymology*
  • Deafness / genetics*
  • Electron Transport
  • Humans
  • Membrane Potential, Mitochondrial
  • Mitochondria / enzymology*
  • Mitochondria / pathology*
  • Mitochondrial Proteins / metabolism
  • Molecular Dynamics Simulation
  • Mutation / genetics*
  • Nucleic Acid Conformation
  • Nucleic Acid Denaturation
  • RNA Stability / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism
  • RNA, Transfer, His / chemistry
  • RNA, Transfer, His / genetics*
  • RNA, Transfer, His / metabolism
  • Reactive Oxygen Species / metabolism
  • Subcellular Fractions / metabolism


  • DNA, Mitochondrial
  • Mitochondrial Proteins
  • RNA, Messenger
  • RNA, Ribosomal
  • RNA, Transfer, His
  • Reactive Oxygen Species
  • Adenosine Triphosphate
  • Amino Acyl-tRNA Synthetases
  • HARS2 protein, human

Associated data

  • PDB/3TUP