The University of Cambridge, Department of Chemistry, Lensfield Road, Cambridge, CB2 1EW, UK.
Angew Chem Int Ed Engl. 2016 Jul 25;55(31):8958-61. doi: 10.1002/anie.201603562. Epub 2016 Jun 29.
RNA G-quadruplex (rG4) structures are of fundamental importance to biology. A novel approach is introduced to detect and structurally map rG4s at single-nucleotide resolution in RNAs. The approach, denoted SHALiPE, couples selective 2'-hydroxyl acylation with lithium ion-based primer extension, and identifies characteristic structural fingerprints for rG4 mapping. We apply SHALiPE to interrogate the human precursor microRNA 149, and reveal the formation of an rG4 structure in this non-coding RNA. Additional analyses support the SHALiPE results and uncover that this rG4 has a parallel topology, is thermally stable, and is conserved in mammals. An in vitro Dicer assay shows that this rG4 inhibits Dicer processing, supporting the potential role of rG4 structures in microRNA maturation and post-transcriptional regulation of mRNAs.
RNA 四链体 (rG4) 结构对生物学具有重要意义。本文介绍了一种新的方法，可在单核苷酸分辨率下检测和结构映射 RNA 中的 rG4。该方法称为 SHALiPE，它结合了选择性 2'-羟基酰化和基于锂离子的引物延伸，为 rG4 作图识别出特征结构指纹。我们应用 SHALiPE 来研究人类前体 microRNA 149，并揭示了该非编码 RNA 中 rG4 结构的形成。进一步的分析支持了 SHALiPE 的结果，并揭示了该 rG4 具有平行拓扑结构，热稳定，并且在哺乳动物中保守。体外 Dicer 测定表明，该 rG4 抑制 Dicer 加工，支持 rG4 结构在 microRNA 成熟和 mRNA 转录后调控中的潜在作用。
