Geneticin reduces mRNA stability

PLoS One. 2022 Jul 28;17(7):e0272058. doi: 10.1371/journal.pone.0272058. eCollection 2022.

Abstract

Messenger RNA (mRNA) translation can lead to higher rates of mRNA decay, suggesting the ribosome plays a role in mRNA destruction. Furthermore, mRNA features, such as codon identities, which are directly probed by the ribosome, correlate with mRNA decay rates. Many amino acids are encoded by synonymous codons, some of which are decoded by more abundant tRNAs leading to more optimal translation and increased mRNA stability. Variable translation rates for synonymous codons can lead to ribosomal collisions as ribosomes transit regions with suboptimal codons, and ribosomal collisions can promote mRNA decay. In addition to different translation rates, the presence of certain codons can also lead to higher or lower rates of amino acid misincorporation which could potentially lead to protein misfolding if a substituted amino acid fails to make critical contacts in a structure. Here, we test whether Geneticin-G418, an aminoglycoside antibiotic known to promote amino acid misincorporation-affects mRNA stability. We observe that G418 decreases firefly luciferase mRNA stability in an in vitro translation system and also reduces mRNA stability in mouse embryonic stem cells (mESCs). G418-sensitive mRNAs are enriched for certain optimal codons that contain G or C in the wobble position, arguing that G418 blunts the stabilizing effects of codon optimality.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / genetics
  • Amino Acids / metabolism
  • Animals
  • Codon / genetics
  • Gentamicins
  • Mice
  • Protein Biosynthesis*
  • RNA Stability*
  • RNA, Messenger / metabolism

Substances

  • Amino Acids
  • Codon
  • Gentamicins
  • RNA, Messenger
  • antibiotic G 418

Grants and funding

Our work was funded with financial support from the Thomas F. and Kate Miller Jeffress Memorial Trust. The Jeffress Memorial Trust did not provide salaries to any of the authors for this study. Please note that the funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.