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Related Experiment Videos

Mitochondrial tRNA(Trp)s Imply a Eubacterial Origin.

Li Chen1, You-Xin Jin, De-Bao Wang

  • 1State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. yxjin@sunm.shcnc.ac.cn

Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao Acta Biochimica Et Biophysica Sinica
|July 6, 2002
PubMed
Summary

Mitochondrial transfer RNA (tRNA) genes from rice, humans, and yeast were recognized by bacterial enzymes but not rat enzymes. This suggests a eubacterial origin for mitochondrial tRNA(Trp)s.

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Area of Science:

  • Molecular Biology
  • Evolutionary Biology
  • Genetics

Background:

  • Mitochondrial transfer RNAs (tRNAs) play crucial roles in protein synthesis within mitochondria.
  • The evolutionary origins of mitochondrial components, including tRNAs, are of significant scientific interest.
  • Understanding tRNA structure-function relationships provides insights into cellular processes and evolutionary history.

Purpose of the Study:

  • To investigate the substrate specificity of bacterial and mammalian tryptophanyl-tRNA synthetases (TrpRS) towards mitochondrial tRNA(Trp)s.
  • To explore the evolutionary implications of interactions between bacterial TrpRS and mitochondrial tRNAs from different species.
  • To characterize the kinetic parameters of tryptophanylation for various mitochondrial tRNA(Trp)s using Bacillus subtilis TrpRS.

Main Methods:

Related Experiment Videos

  • Construction and in vitro transcription of mitochondrial tRNA(Trp) genes from Oryza sativa, Homo sapiens, and Saccharomyces cerevisiae.
  • Enzymatic assays to assess tryptophanylation activity of Bacillus subtilis and rat liver TrpRS on mitochondrial tRNA(Trp) transcripts.
  • Kinetic analysis (kcat/Km) to determine binding capacity and catalytic efficiency of B. subtilis TrpRS for different tRNA(Trp) substrates.

Main Results:

  • In vitro transcripts of mitochondrial tRNA(Trp)s from O. sativa, H. sapiens, and S. cerevisiae were successfully tryptophanylated by Bacillus subtilis TrpRS.
  • These mitochondrial tRNA(Trp)s were not recognized or catalyzed by rat liver TrpRS.
  • Kinetic assays revealed that B. subtilis TrpRS exhibited half the binding capacity for mitochondrial tRNA(Trp)s compared to wild-type B. subtilis tRNA(Trp).
  • Catalytic efficiency (kcat/Km) was significantly reduced for mitochondrial tRNA(Trp)s from O. sativa (400-fold) and S. cerevisiae (1200-fold) compared to B. subtilis tRNA(Trp).

Conclusions:

  • The substrate specificity of Bacillus subtilis TrpRS towards mitochondrial tRNA(Trp)s suggests functional conservation.
  • The differential recognition by bacterial versus mammalian TrpRS supports the endosymbiotic theory and points to a eubacterial origin of mitochondrial tRNA(Trp)s.
  • These findings contribute to understanding the evolutionary trajectory of mitochondrial gene expression machinery.