Single-molecule imaging reveals translation-dependent destabilization of mRNAs

Mol Cell. 2023 Feb 16;83(4):589-606.e6. doi: 10.1016/j.molcel.2023.01.013. Epub 2023 Feb 1.

Abstract

The relationship between mRNA translation and decay is incompletely understood, with conflicting reports suggesting that translation can either promote decay or stabilize mRNAs. The effect of translation on mRNA decay has mainly been studied using ensemble measurements and global transcription and translation inhibitors, which can have pleiotropic effects. We developed a single-molecule imaging approach to control the translation of a specific transcript that enabled simultaneous measurement of translation and mRNA decay. Our results demonstrate that mRNA translation reduces mRNA stability, and mathematical modeling suggests that this process is dependent on ribosome flux. Furthermore, our results indicate that miRNAs mediate efficient degradation of both translating and non-translating target mRNAs and reveal a predominant role for mRNA degradation in miRNA-mediated regulation. Simultaneous observation of translation and decay of single mRNAs provides a framework to directly study how these processes are interconnected in cells.

Keywords: NMD; mRNA decay; miRNA; single molecule; translation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • MicroRNAs* / genetics
  • MicroRNAs* / metabolism
  • Nonsense Mediated mRNA Decay*
  • Protein Biosynthesis
  • RNA Stability / genetics
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Ribosomes / genetics
  • Ribosomes / metabolism
  • Single Molecule Imaging

Substances

  • RNA, Messenger
  • MicroRNAs