Specific complexes of protein and RNA carry out many essential biological functions, including RNA processing, RNA turnover, RNA folding, as well as the translation of genetic information from mRNA into protein sequences. Messenger RNA (mRNA) decay is now emerging as an important control point and a major contributor to gene expression. Continuing identification of the protein factors and cofactors, and mRNA instability elements, responsible for mRNA decay allow researchers to build a comprehensive picture of the highly orchestrated processes involved in mRNA decay and its regulation.

Key Features

Covers the difference in processing of mRNA between eukaryotes, bacteria and archea. Benefit: Processing of mRNA differs greatly between eukaryotes, bacteria and archea and this affords researchers readily reproducible techniques to understand and study the molecular pathogenesis of disease. Expert researchers introduce the most advanced technologies and techniques to identify mRNA processing, transport, localization and turnover which are central to the process of gene expression. Benefit: Keeps MIE buyers and online subscribers up-to-date with the latest research Offers step by step lab instructions including necessary equipment and reagents. Benefit: Provides tried and tested techniques which eliminate searching through many different sources. Tested techniques are trustworthy and avoid pitfalls so the same mistakes are not made over and over.


Researchers in biochemistry, cell and molecular biology, genetics

Table of Contents

Part I Bacteria Analysis of RNA decay, including polyadenylation, in mutant E. coli Analyzing the decay of stable RNAs in E. coli Genomic analyses of mRNA decay in E. coli using microarrays Methods to co-immunopurify degradative activities in E. coli Proteomics approach (affinity purification) for analyzing degradosome composition in E. coli Characterizing decay in vitro, including structural studies Defining RNA degradative activities in pathogenic bacteria (e.g., Studying tm-mediated surveillance and nonstop mRNA decay Defining small RNA function in RNA decay Characterizing mRNA destabilization mediated by Hfq-binding noncoding RNAs Part II Archaea Methods for the global analysis of mRNA stability in Archaea In vivo and in vitro studies of degradative activities in Archaea Structural studies of degradative activities using Archaea Part III Eukaryotes Dcp2/Dcp1 single-step kinetics Dcp2/Dcp1 and DCPS activity assays Purification and analyses of decapping activtors in Saccharomyces Reconstitution and analyses of the human Lsm complex Analyses of deadenylation in S. cerevisiae in vitro Deadenylation in mammalian-cell extracts Cell-free deadenylation assays using Drosophila embryos Cytoplasmic deadenylation assays using Xenopus oocytes Activity and structural analyses of the reconstituted eukaryotic RNA exosome Biochemical studies of the eukaryotic exosome Assays of 5?-3? exonucleases in Saccharomyces cerevisiae mRNA half-life measurements in Saccharomyces cerevisiae (all methods, including ts RNA polymerase) Genome-wide analyses of mRNA stability in Sacc


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© 2008
Academic Press
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Print ISBN:

About the editors

Lynne E. Maquat

Affiliations and Expertise

University of Rochester, NY, USA

Cecilia Arraiano

Affiliations and Expertise

Universidade Nova de Lisboa, Portugal


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