Chapter 5 Regulated Deadenylation In Vitro

Aaron C. Goldstrohm, Brad A. Hook, Marvin Wickens

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Scopus citations

Abstract

The 3′-poly(A) tail, found on virtually all mRNAs, is enzymatically shortened by a process referred to as "deadenylation." Deadenylation is a widespread means of controlling mRNA stability and translation. The enzymes involved-so-called deadenylases-are surprisingly diverse. They are controlled by RNA sequences commonly found in 3′-untranslated regions (UTRs), which bind regulatory factors. Both RNA-binding proteins and microRNAs accelerate deadenylation of specific mRNAs. In some cases, regulators enhance deadenylation by binding to and recruiting specific deadenylases to the target mRNA. The many hundreds of potential regulators encoded in mammalian genomes (both RNA-binding proteins and microRNAs) and the numerous deadenylases, coupled with the many potential regulatory sites represented in 3′ UTRs of mRNAs, provide fertile ground for regulated deadenylation. Recent global studies of poly(A) regulation support this conclusion. Biochemical and genetic approaches will be essential for exploring regulated deadenylation. The methods we describe focus on the reconstruction in vitro of regulated deadenylation with purified components from yeast. We discuss broadly the strategies, problems, and history of in vitro deadenylation systems. We combine this with a more detailed discussion of the purification, activity, and regulation of the Saccharomyces cerevisiae Ccr4p-Pop2p deadenylase complex and its regulation by PUF (Pumilio and Fem-3 binding factor) RNA-binding proteins.

Original languageEnglish (US)
Title of host publicationRNA Turnover in Eukaryotes
Subtitle of host publicationNucleases, Pathways and Anaylsis of mRNA Decay
EditorsLynne Maquat, Megerditch Kiledjian
Pages77-106
Number of pages30
DOIs
StatePublished - Dec 24 2008

Publication series

NameMethods in Enzymology
Volume448
ISSN (Print)0076-6879

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