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A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues
08:56

A Method for Measuring RNA N6-methyladenosine Modifications in Cells and Tissues

Published on: December 5, 2016

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N(6)-methyladenosine in mRNA disrupts tRNA selection and translation-elongation dynamics

Junhong Choi1,2, Ka-Weng Ieong3, Hasan Demirci4,5

  • 1Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA.

Nature Structural & Molecular Biology
|January 12, 2016
PubMed

Insights

The abundant mRNA modification N(6)-methylation of adenosine (m(6)A) hinders translation by impeding tRNA accommodation and elongation, impacting decoding dynamics. This chemical modification alters how ribosomes process mRNA during protein synthesis.

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • N(6)-methylation of adenosine (m(6)A) is the most prevalent mRNA modification.
  • Its functional role during the translation process remains largely unexplored.

Purpose of the Study:

  • To investigate the impact of m(6)A on mRNA decoding and translation dynamics.
  • To elucidate the mechanistic basis of m(6)A's effect on protein synthesis.

Main Methods:

  • Utilized bulk kinetic and single-molecule assays to study translation in an Escherichia coli system.
  • Employed X-ray crystallography of Thermus thermophilus ribosomal complexes to analyze m(6)A-U base pairing.

Main Results:

  • m(6)A modification on mRNA acts as a barrier to tRNA accommodation and translation elongation.
  • The m(6)A-codon and cognate tRNA interaction mimics that of a near-cognate codon and tRNA.
  • The effect of m(6)A is context-dependent, influenced by its position and the translation accuracy level.

Conclusions:

  • mRNA chemical modifications, specifically m(6)A, significantly alter translational dynamics.
  • m(6)A influences the efficiency and accuracy of protein synthesis by modulating ribosome-mediated decoding.

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