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Chapter 1: Methods to Study No-Go mRNA Decay in Saccharomyces cerevisiae

Doma, Meenakshi K. (2008) Chapter 1: Methods to Study No-Go mRNA Decay in Saccharomyces cerevisiae. In: RNA Turnover in Eukaryotes: Analysis of Specialized and Quality Control RNA Decay Pathways. Methods in Enzymology. No.449. Academic Press , San Diego, CA, pp. 3-21. ISBN 9780123745842. https://resolver.caltech.edu/CaltechAUTHORS:20090728-094003163

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Abstract

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.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1016/S0076-6879(08)02401-4DOIUNSPECIFIED
Additional Information:© 2008 Elsevier. Available online 10 February 2009. I thank Roy Parker and Kristian Baker for discussions and critical review of the manuscript. All work described in this chapter has been done at the Department of Molecular and Cellular Biology and HHMI, University of Arizona, Tucson, Arizona, and supported by funds from the Howard Hughes Medical Institute and the National Institute of Health.
Funders:
Funding AgencyGrant Number
Howard Hughes Medical InstituteUNSPECIFIED
NIHUNSPECIFIED
Subject Keywords:Genes: PGK1, eRF3, UPF1, UPF2; Compounds: galactose, lithium, nitrogen; Biogroups: cytoplasm, mRNA cleavage, mRNA stability, exonuclease, mRNA transcription, protein stability, RNA interference, signal recognition particle binding
Series Name:Methods in Enzymology
Issue or Number:449
Record Number:CaltechAUTHORS:20090728-094003163
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20090728-094003163
Official Citation:Meenakshi K. Doma, Chapter 1 Methods to Study No[hyphen (true graphic)]Go mRNA Decay in Saccharomyces cerevisiae, In: Lynne E. Maquat and Megerditch Kiledjian, Editor(s), Methods in Enzymology, Academic Press, 2008, Volume 449, RNA Turnover in Eukaryotes: Analysis of Specialized and Quality Control RNA Decay Pathways, Pages 3-21, ISSN 0076-6879, ISBN 9780123745842, DOI: 10.1016/S0076-6879(08)02401-4. (http://www.sciencedirect.com/science/article/B7CV2-4VK64G8-6/2/0581a1326e7539733e355a12d2f2f090)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:14694
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:01 Sep 2009 21:50
Last Modified:03 Oct 2019 00:51

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