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Translatome profiling: methods for genome-scale analysis of mRNA translation

King, HA and Gerber, AP (2014) Translatome profiling: methods for genome-scale analysis of mRNA translation Brief Funct Genomics.

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Abstract

During the past decade, there has been a rapidly increased appreciation of the role of translation as a key regulatory node in gene expression. Thereby, the development of methods to infer the translatome, which refers to the entirety of mRNAs associated with ribosomes for protein synthesis, has facilitated the discovery of new principles and mechanisms of translation and expanded our view of the underlying logic of protein synthesis. Here, we review the three main methodologies for translatome analysis, and we highlight some of the recent discoveries made using each technique. We first discuss polysomal profiling, a classical technique that involves the separation of mRNAs depending on the number of bound ribosomes using a sucrose gradient, and which has been combined with global analysis tools such as DNA microarrays or high-throughput RNA sequencing to identify the RNAs in polysomal fractions. We then introduce ribosomal profiling, a recently established technique that enables the mapping of ribosomes along mRNAs at near-nucleotide resolution on a global scale. We finally refer to ribosome affinity purification techniques that are based on the cell-type-specific expression of tagged ribosomal proteins, allowing the capture of translatomes from specialized cells in organisms. We discuss the advantages and disadvantages of these three main techniques in the pursuit of defining the translatome, and we speculate about future developments.

Item Type: Article
Divisions : Faculty of Health and Medical Sciences > School of Biosciences and Medicine > Department of Microbial and Cellular Sciences
Authors :
AuthorsEmailORCID
King, HAUNSPECIFIEDUNSPECIFIED
Gerber, APUNSPECIFIEDUNSPECIFIED
Date : 6 November 2014
Identification Number : 10.1093/bfgp/elu045
Additional Information : © The Author 2014. Published by Oxford University Press. All rights reserved.This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Brief Funct Genomics following peer review. The definitive publisher-authenticated version is available online at http://dx.doi.org/10.1093/bfgp/elu045.
Depositing User : Symplectic Elements
Date Deposited : 02 Dec 2014 13:55
Last Modified : 06 Nov 2015 02:08
URI: http://epubs.surrey.ac.uk/id/eprint/806765

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