Chapter 1. Methods to study no-go mRNA decay in Saccharomyces cerevisiae

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Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains
12:21

Measurement of mRNA Decay Rates in Saccharomyces cerevisiae Using rpb1-1 Strains

Published on: December 13, 2014

Meenakshi K Doma1

  • 1Division of Biology and Howard Hughes Medical Institute, California Institute of Technology, Pasadena, California, USA.

Methods in Enzymology
|February 14, 2009
PubMed

Insights

This study introduces no-go mRNA decay (NGD), a novel RNA quality control pathway in yeast that degrades aberrant mRNAs with translation elongation defects. Methods are provided to study NGD and identify similar pathways in other organisms.

Area of Science:

  • Molecular Biology
  • Genetics
  • RNA Biology

Background:

  • Eukaryotic cells possess mRNA surveillance systems to degrade aberrant mRNAs, ensuring gene expression quality control.
  • Nonsense-mediated mRNA decay and nonstop mRNA decay are known cytoplasmic pathways targeting translation errors.
  • A novel pathway, no-go mRNA decay (NGD), has been identified in yeast for mRNAs with translation elongation defects.

Purpose of the Study:

  • To describe methods for studying no-go mRNA decay (NGD) in Saccharomyces cerevisiae.
  • To provide procedures for creating translation elongation pauses and assaying NGD substrates.
  • To enable characterization of NGD-dependent endonucleolytic cleavage and identification of novel RNA quality control pathways.

Main Methods:

  • Induction of efficient translation elongation pauses in yeast mRNA.
  • Assay development for evaluating decay characteristics of NGD substrates.
  • Characterization of endonucleolytic cleavage specific to NGD pathways.

Main Results:

  • Established methods for creating and analyzing translation elongation pauses.
  • Developed assays to measure mRNA decay rates influenced by NGD.
  • Characterized the endonucleolytic cleavage mechanism central to NGD.

Conclusions:

  • The described methods facilitate the study of no-go mRNA decay (NGD) in yeast.
  • The principles can be adapted to discover and analyze new RNA quality control mechanisms.
  • This research enhances understanding of eukaryotic mRNA surveillance pathways.

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