Identification of RNA Oligonucleotides Binding to Several Proteins from Potential G-Quadruplex Forming Regions in Transcribed Pre-mRNA.
Identification of RNA Oligonucleotides Binding to Several Proteins from Potential G-Quadruplex Forming Regions in Transcribed Pre-mRNA.
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DOI：
10.3390/molecules201119733
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发表时间：
2015-11-23
期刊：
影响因子：
--
通讯作者：
Ikebukuro K
中科院分区：
文献类型：
--
作者：
Saito T;Yoshida W;Yokoyama T;Abe K;Ikebukuro K
G-四链体 (G4s) 是由富含鸟嘌呤的序列形成的非典型 DNA/RNA 结构。最近，G4不仅在适体中被发现，还在基因组DNA和转录RNA中被发现。在这项研究中，我们通过关注位于前体 mRNA 内的 G4 形成 RNA，鉴定出了充当适体的新 RNA 寡核苷酸。我们发现，VEGFA 5' UTR 和第一个内含子中的 G4 以高亲和力与 VEGFA 基因编码的蛋白质 VEGF165 结合。此外，位于PDGFA和PDGFB内含子内的G4形成RNA分别与PDGF-AA和PDGF-BB结合，表明前体mRNA中的G4可能是适体。据报道，假定的 G4 形成 RNA 序列位于大多数基因的某些部分，因此我们的适体鉴定策略可适用于其他蛋白质。据报道，5'UTR 中的一些形成 G4 的 RNA 参与翻译控制；然而，G4 形成切除的内含子 RNA 功能之前尚未被揭示。因此，这些发现不仅有助于RNA适体的鉴定，而且还为位于内含子RNA序列内的G4形成RNA的生物学功能提供了新的见解。
G-quadruplexes (G4s) are noncanonical DNA/RNA structures formed by guanine-rich sequences. Recently, G4s have been found not only in aptamers but also in the genomic DNA and transcribed RNA. In this study, we identified new RNA oligonucleotides working as aptamers by focusing on G4-forming RNAs located within the pre-mRNA. We showed that the G4 in the 5′ UTR and first intron of VEGFA bound to the protein encoded in VEGFA gene, VEGF165, with high affinity. Moreover, G4-forming RNAs located within the PDGFA and the PDGFB introns bound to PDGF-AA and PDGF-BB, respectively, indicating that G4 in the pre-mRNA could be an aptamer. It had been reported that the putative G4-forming RNA sequences are located in some parts of most genes, thus our strategy for aptamer identification could be applicable to other proteins. It has been reported that some G4-forming RNAs in 5′ UTRs are involved in translation control; however, G4-forming excised intronic RNA function has not been revealed previously. Therefore, these findings could not only contribute to the identification of RNA aptamers but also provide new insights into the biological functioning of G4-forming RNAs located within intronic RNA sequences.
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