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Nucleic Acids Research Advance Access published online on October 22, 2009
Nucleic Acids Research, doi:10.1093/nar/gkp870
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© The Author(s) 2009. Published by Oxford University Press.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

RNA

Sequence context outside the target region influences the effectiveness of miR-223 target sites in the RhoB 3'UTR

Guihua Sun1,2, Haitang Li2 and John J. Rossi1,2,*

1Irell & Manella Graduate School of Biological Science and 2Department of Molecular Biology, Beckman Research Institute of the City of Hope, 1500 E. Duarte Road, Duarte, CA 91010-3000, USA

*To whom correspondence should be addressed. Tel: +1 626 301 8360; Fax: +1 626 301 8271; Email: jrossi{at}coh.org

Received August 7, 2009. Revised September 29, 2009. Accepted September 30, 2009.

MicroRNAs (miRNAs) are 21–22 nucleotide regulatory small RNAs that repress message translation via base-pairing with complementary sequences in the 3' untranslated region (3'UTR) of targeted transcripts. To date, it is still difficult to find a true miRNA target due to lack of a clear understanding of how miRNAs functionally interact with their targeted transcripts for efficient repression. Previous studies have shown that nucleotides 2 to 7 at the 5'-end of a mature miRNA, the ‘seed sequence’, can nucleate miRNA/target interactions. In the current study, we have validated that the RhoB mRNA is a bona fide miR-223 target. We have analyzed the functional activities of two miR223-binding sites within the RhoB 3'UTR. We find that the two miR-223 target sites in the RhoB 3'UTR contribute differentially to the total repression of RhoB translation. Moreover, we demonstrate that some AU-rich motifs located upstream of the distal miRNA-binding site enhance miRNA function, independent of the miRNA target sequences being tested. We also demonstrate that the AU-rich sequence elements are polar, and do not affect the activities of miRNAs whose sites lie upstream of these elements. These studies provide further support for the role of sequences outside of miRNA target region influencing miRNA function.


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