Transcriptome-wide measurement of mRNA polyadenylation state

被引：15

作者：

Beilharz, Traude H. ^{[1
,2
]}

Preiss, Thomas ^{[1
,2
,3
]}

机构：

[1] VCCRI, Div Mol Genet, Sydney, NSW 2010, Australia

[2] Univ New S Wales, St Vincents Clin Sch, Sydney, NSW 2052, Australia

[3] Univ New S Wales, Sch Biotechnol & Biomol Sci, Sydney, NSW 2052, Australia

来源：

METHODS | 2009年 / 48卷 / 03期

基金：

澳大利亚研究理事会; 英国医学研究理事会;

关键词：

Poly(A) tail; Polyadenylation; Deadenylation; Translational control; mRNA decay; Post-transcriptional regulation; Polysome; Microarray; Transcriptome; Proteome; EUKARYOTIC TRANSLATION INITIATION; SACCHAROMYCES-CEREVISIAE; POLY(A) TAIL; DEADENYLASE; DEGRADATION; FRACTIONATION; PROTEINS; REVEALS; COMPLEX; CELLS;

D O I：

10.1016/j.ymeth.2009.02.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The 3' poly(A) tail has important roles throughout the eukaryotic mRNA life cycle. A characteristic aspect of poly(A) tail function is furthermore that it can be modulated by changes in its length. This is in turn a well-recognised cellular means to regulate both, mRNA translation and stability, and a positive correlation has often been found between the efficiency of mRNA translation and the length of its poly(A) tail. Here we describe methodology to measure mRNA polyadenylation state in a transcriptome-wide manner, using separation of cellular mRNA populations on poly(U) sepharose in combination with microarray analysis of the resulting fractions. We further detail methods for bulk and mRNA-specific poly(A) tail length measurements to monitor the efficiency of initial mRNA separation and to verify candidates selected from the microarray data. Although detailed here for the study of yeast mRNAs, these methods are adaptable to the investigation of any cellular context in which poly(A) tail length control is known or suspected to operate. (C) 2009 Elsevier Inc. All rights reserved.

引用

页码：294 / 300

页数：7