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Nucl. Acids Res.-2007-Meijer-e132.pdf (2.23 MB)

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

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posted on 2012-10-24, 09:11 authored by H. A. Meijer, M. Bushell, K. Hill, A. E. Willis, de Moor C. H., P. Jones, Timothy W. Gant
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 ∼25% of the expressed genes have a poly(A) tail of less than 30 residues in a significant percentage of their transcripts.

Funding

This work was funded by Wellcome Trust (076179); Biotechnology and Biological Sciences Research Council (42/G14670). Funding to pay the Open Access publication charges for this article was provided by the Wellcome Trust.

History

Citation

Nucleic Acids Research, 2007, 35 (19)

Published in

Nucleic Acids Research

Publisher

Oxford University Press (OUP)

issn

0305-1048

eissn

1362-4962

Copyright date

2007

Available date

2012-10-24

Publisher version

http://nar.oxfordjournals.org/content/35/19/e132

Language

en

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