Computational identification of biologically functional non-hairpin GC-helices in human Argonaute mRNA

Simon Dornseifer,Georg Sczakiel

doi:10.1186/1471-2105-14-122

Abstract

BackgroundPerfectly formed duplex elements in RNA occur within folding units, often as a part of hairpin motifs which can be reliably predicted by various RNA folding algorithms. Double helices with consecutive Watson-Crick base-pairing may also be formed between distant RNA segments thereby facilitating long-range interactions of long-chain RNA that may be biologically functional. Here we addressed the potential formation of RNA duplex motifs by long-range RNA-RNA interactions of distantly located matching sequence elements of a single long-chain RNA.ResultsWe generated a Python-based software tool that identifies consecutive RNA duplex elements at any given length and nucleotide content formed by distant sequences. The software tool, dubbed RNAslider, is built on the theoretical RNA structure prediction algorithm Mfold. Source code and sample data sets are available on demand. We found that a small ratio of human genes including the Argonaute (Ago)-like gene family encode mRNAs containing highly GC-rich non-hairpin duplex elements (GC-helix) of equal to or more than 8 base pairs in length and we provide experimental evidence for their biological significance.ConclusionGC-helices are observed preferentially within the 5′-region of mRNAs in an evolutionarily conserved fashion indicating their potential biological role. This view is supported experimentally by post-transcriptional regulation of gene expression of a fusion transcript containing 5′-sequences of human mRNAAgo2 harbouring GC-helices and down-stream coding sequences of Renilla luciferase.

Highlights

Formed duplex elements in RNA occur within folding units, often as a part of hairpin motifs which can be reliably predicted by various RNA folding algorithms
We studied the occurrence of non-hairpin duplex elements including GC-rich or AU-rich helices in a library of human mRNA sequences by using this software tool
The results were screened for duplex elements, i.e. helices according to the definitions described in 2.1

Summary

Introduction

Formed duplex elements in RNA occur within folding units, often as a part of hairpin motifs which can be reliably predicted by various RNA folding algorithms. Double helices with consecutive Watson-Crick base-pairing may be formed between distant RNA segments thereby facilitating long-range interactions of long-chain RNA that may be biologically functional. Preliminary experimental studies in our laboratory indicate specific binding of RNA sequences surrounding the AUG start codon of the Ago mRNA with proteins involved in RNA interference (Sczakiel, unpublished data). This attracted us to study the 50-UTR of mRNAAgo with regard to potential RNA cis-elements. Systematic computational and phylogenetic studies as well as experimental evidence in a mammalian cell system support the view that GC-rich duplex motifs formed by distant RNA segments, unlike typical hairpin elements, could bear biological functions

Methods

Results

Conclusion