A novel method for poly(A) fractionation reveals a large population of mRNAs with a short poly(A) tail in mammalian cells

被引：80

作者：

Meijer, Hedda A.

Bushell, Martin

Hill, Kirsti

Gant, Timothy W.

Willis, Anne E.

Jones, Peter

de Moor, Cornelia H. ^{[1
]}

机构：

[1] Univ Nottingham, Sch Pharm, Ctr Biomol Sci, RNA Biol Grp, Nottingham NG7 2RD, England

[2] Queens Med Ctr, Sch Biomed Sci, Ctr Biochem & Cell Biol, Nottingham NG7 2UH, England

[3] Univ Leicester, MRC Toxicol Unit, Leicester LE1 9HN, Leics, England

来源：

NUCLEIC ACIDS RESEARCH | 2007年 / 35卷 / 19期

基金：

英国惠康基金;

关键词：

D O I：

10.1093/nar/gkm830

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The length of the poly(A) tail of an mRNA plays an important role in translational efficiency, mRNA stability and mRNA degradation. Regulated polyadenylation and deadenylation of specific mRNAs is involved in oogenesis, embryonic development, spermatogenesis, cell cycle progression and synaptic plasticity. Here we report a new technique to analyse the length of poly(A) tails and to separate a mixed population of mRNAs into fractions dependent on the length of their poly(A) tails. The method can be performed on crude lysate or total RNA, is fast, highly reproducible and minor changes in poly(A) tail length distribution are easily detected. We validated the method by analysing mRNAs known to undergo cytoplasmic polyadenylation during Xenopus laevis oocyte maturation. We then separated RNA from NIH3T3 cells into two fractions with short and long poly(A) tails and compared them by microarray analysis. In combination with the validation experiments, the results indicate that similar to 25% of the expressed genes have a poly(A) tail of less than 30 residues in a significant percentage of their transcripts.

引用

页数：13

共 51 条

[1] Widespread use of poly(A) tail length control to accentuate expression of the yeast transcriptome [J].