结合HCV IRES的RNA适体的结构抑制分析。

Kunio Kikuchi, Takuya Umehara, Kotaro Fukuda, Joonsung Hwang, Atsushi Kuno, Tsunemi Hasegawa, Satoshi Nishikawa
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引用次数: 13

摘要

HCV的翻译始于5'非翻译区的内部核糖体进入位点(IRES), IRES在HCV株中保存良好。我们利用生物素化寡核苷酸探针开发了一种新的选择策略,获得了分别结合HCV IRES结构域II和结构域III-IV的RNA适配体。选择的适体通过RNA-RNA相互作用特异性结合到目标序列。这些适体在体外抑制ires依赖性翻译。特别是结合结构域IIId的3-07适体表现出较强的抑制作用。通过诱变、RNase作图和结合动力学分析了这些适体的结构/功能关系。
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Structure-inhibition analysis of RNA aptamers that bind to HCV IRES.
The translation of HCV starts at the internal ribosomal entry site (IRES) within the 5' untranslated region and IRES is well-conserved in HCV strains. We developed a novel selection strategy using biotinylated oligonucleotide probe and obtained RNA aptamers that bind HCV IRES domain II and domain III-IV, respectively. Selected aptamers specifically bound to target sequence via RNA-RNA interactions. These aptamers inhibited IRES-depend translation in vitro. Especially, 3-07 aptamer, which bound domain IIId, showed strong inhibition. Structure/function relationship of these aptamers was analyzed by mutagenesis, RNase mapping and binding kinetics.
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