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3'非翻译区G-四链体调节微小RNA结合。

3' UTR G-quadruplexes regulate miRNA binding.

作者信息

Rouleau Samuel, Glouzon Jean-Pierre Sehi, Brumwell Andrea, Bisaillon Martin, Perreault Jean-Pierre

机构信息

Département de Biochimie, Pavillon de Recherche Appliquée sur le Cancer, Université de Sherbrooke, Sherbrooke (Québec), Canada J1E 4K8.

出版信息

RNA. 2017 Aug;23(8):1172-1179. doi: 10.1261/rna.060962.117. Epub 2017 May 4.

DOI:10.1261/rna.060962.117
PMID:28473452
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5513062/
Abstract

MicroRNAs (miRNAs) are small noncoding RNAs that repress the translation of their target genes. It has previously been shown that a target's availability to miRNA can be affected by its structure. G-quadruplexes (G4) are noncanonical structures adopted by G-rich nucleic acids that have been shown to have multiple biological functions. In this study, whether or not G4 structures' presence in the 3' UTRs of mRNAs can hinder miRNA binding was investigated. Putative G4 overlapping with predicted miRNAs' binding sites was searched for, and 44,294 hits were found in humans. The FADS2 mRNA/mir331-3p pair was selected as a model example. In-line probing and G4-specific fluorescent ligand experiments binding were performed and confirmed the presence of a G4 near the predicted miRNA binding site. Subsequent luciferase assays showed that the presence of the G4 prevents the binding of mir331-3p in cellulo. Together, these results served as proof of concept that a G4 structure present in a 3' UTR sequence should be taken into consideration when predicting miRNA binding sites.

摘要

微小RNA（miRNA）是一类小的非编码RNA，可抑制其靶基因的翻译。此前已有研究表明，靶标对miRNA的可及性会受其结构影响。G-四链体（G4）是富含鸟嘌呤的核酸所形成的非经典结构，已被证明具有多种生物学功能。在本研究中，我们探究了mRNA的3'非翻译区（UTR）中G4结构的存在是否会阻碍miRNA结合。我们搜索了与预测的miRNA结合位点重叠的假定G4，在人类中发现了44294个匹配结果。选择FADS2 mRNA/mir331-3p对作为典型示例。进行了在线探针实验和G4特异性荧光配体结合实验，证实了在预测的miRNA结合位点附近存在G4。随后的荧光素酶检测表明，G4的存在会阻止mir331-3p在细胞内的结合。总之，这些结果证明了在预测miRNA结合位点时，应考虑3'UTR序列中存在的G4结构。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/3963c609ac60/1172f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/2a3c5e1b1074/1172f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/d19a02585127/1172f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/daeaa05193b2/1172f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/3963c609ac60/1172f04.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/2a3c5e1b1074/1172f01.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/d19a02585127/1172f02.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/daeaa05193b2/1172f03.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b293/5513062/3963c609ac60/1172f04.jpg

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