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PMID: 17933768 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Validation Study

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

Nucleic acids research ·Vol. 35 ·No. 19 ·2007-00-00 ·Pages e132

Meijer HA, Bushell M, Hill K, Gant TW, Willis AE, Jones P, de Moor CH

Abstract

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 approximately 25% of the expressed genes have a poly(A) tail of less than 30 residues in a significant percentage of their transcripts.

MeSH Terms
3' Untranslated Regions/chemistry Animals Chromatography, Affinity/methods Mice NIH 3T3 Cells Oligonucleotide Array Sequence Analysis Oocytes/metabolism Poly A/analysis Polyadenylation RNA, Messenger/isolation & purification Xenopus laevis
Chemicals
3' Untranslated Regions RNA, Messenger Poly A
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Meijer Hedda A
RNA Biology Group, Centre for Biomolecular Sciences, School of Pharmacy, University of Nottingham, Nottingham, UK.
Bushell Martin
Hill Kirsti
Gant Timothy W
Willis Anne E
Jones Peter
de Moor Cornelia H
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkm830
Published
2007-00-00
Epub
2007-00-11
Pages
e132
Language
English
Region
England
NLM ID
0411011
PMCID
PMC2095794
Subset
IM
Grants
Wellcome Trust · 076179 · United Kingdom
External Links
PubMed source DOI: 10.1093/nar/gkm830 PMC Full Text View journal details
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