Mouse let-7 miRNA populations exhibit RNA editing that is constrained in the 5′-seed/cleavage/anchor regions and stabilize predicted mmu-let-7a:mRNA duplexes

Jeffrey G. Reid, Ankur K. Nagaraja, Francis C. Lynn, Rafal B. Drabek, Donna M. Muzny, Chad A. Shaw, Michelle K. Weiss, Arash O. Naghavi, Mahjabeen Khan, Huifeng Zhu, Jayantha Tennakoon, Gemunu H. Gunaratne, David Corry, Jonathan Miller, Michael T. McManus, Michael S. German, Richard A. Gibbs, Martin M. Matzuk, Preethi H. Gunaratne

Research output: Contribution to journalArticle

79 Scopus citations

Abstract

Massively parallel sequencing of millions of <30-nt RNAs expressed in mouse ovary, embryonic pancreas (E14.5), and insulin-secreting beta-cells (βTC-3) reveals that ∼50% of the mature miRNAs representing mostly the mmu-let-7 family display internal insertion/deletions and substitutions when compared to precursor miRNA and the mouse genome reference sequences. Approximately, 12%-20% of species associated with mmu-let-7 populations exhibit sequence discrepancies that are dramatically reduced in nucleotides 3-7 (5′-seed) and 10-15 (cleavage and anchor sites). This observation is inconsistent with sequencing error and leads us to propose that the changes arise predominantly from post-transcriptional RNA-editing activity operating on miRNA:target mRNA complexes. Internal nucleotide modifications are most enriched at the ninth nucleotide position. A common ninth base edit of U-to-G results in a significant increase in stability of down-regulated let-7a targets in inhibin-deficient mice (Inha-/-). An excess of U-insertions (14.8%) over U-deletions (1.5%) and the presence of cleaved intermediates suggest that a mammalian TUTase (terminal uridylyl transferase) mediated dUTP-dependent U-insertion/U-deletion cycle may be a possible mechanism. We speculate that mRNA target site-directed editing of mmu-let-7a duplex-bulges stabilizes "loose" miRNA:mRNA target associations and functions to expand the target repertoire and/or enhance mRNA decay over translational repression. Our results also demonstrate that the systematic study of sequence variation within specific RNA classes in a given cell type from millions of sequences generated by next-generation sequencing (NGS) technologies ("intranomics") can be used broadly to infer functional constraints on specific parts of completely uncharacterized RNAs.

Original languageEnglish (US)
Pages (from-to)1571-1581
Number of pages11
JournalGenome Research
Volume18
Issue number10
DOIs
StatePublished - Oct 2008

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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