Send to

Choose Destination
See comment in PubMed Commons below
Methods Enzymol. 2008;449:3-21. doi: 10.1016/S0076-6879(08)02401-4.

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

Author information

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


In eukaryotic cells, conserved mRNA surveillance systems target and degrade aberrant mRNAs, eliminating translation errors that occur during protein synthesis and thereby imposing quality control of gene expression. Two such cytoplasmic quality control systems, nonsense-mediated mRNA decay and nonstop mRNA decay, have evolved to target mRNAs with aberrancies in translation. A third novel quality control system has been identified for yeast mRNAs with defects in translation elongation due to strong translation pause sites. This subset of mRNAs with ribosome pause sites is recognized and targeted for degradation by an endonucleolytic cleavage in a process referred to as no-go mRNA decay (NGD). The methods described herein are designed to aid in the study of NGD in Saccharomyces cerevisiae. They include procedures to create an efficient translation elongation pause, assay decay characteristics of NGD substrates, and characterize NGD-dependent endonucleolytic cleavage of mRNA. The logic of the design and methods described can be modulated and used for the identification and analysis of novel RNA quality control pathways in other organisms.

[Indexed for MEDLINE]
PubMed Commons home

PubMed Commons

How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Support Center