In vitro selection of RNA aptamers against the HCV NS3 helicase domain.
In vitro selection of RNA aptamers against the HCV NS3 helicase domain.
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DOI：
10.1089/1545457041526335
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发表时间：
2004-08
期刊：
影响因子：
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通讯作者：
F. Nishikawa;K. Funaji;K. Fukuda;S. Nishikawa
中科院分区：
文献类型：
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作者：
F. Nishikawa;K. Funaji;K. Fukuda;S. Nishikawa
丙型肝炎病毒 (HCV) 的非结构蛋白 3 (NS3) 具有两个不同的结构域：蛋白酶和解旋酶，这两个结构域对于 HCV 增殖至关重要。因此，NS3被认为是抗HCV治疗的靶点。为了研究 NS3 解旋酶结构域的 RNA 适体，我们针对 HCV NS3 解旋酶结构域进行了体外选择。八代后获得的RNA适体具有5'延伸的单链区域和茎环区域的保守序列(5'-GGA(U/C)GGAGCC-3')。适体 5 在体外表现出强烈的解旋酶活性抑制作用。缺失和诱变分析阐明保守的茎环很重要并且整个结构是解旋酶抑制所必需的。我们比较了HCV适体5和3'+-UTR对解旋酶活性的抑制作用。
Nonstructural protein 3 (NS3) of hepatitis C virus (HCV) has two distinct domains, protease and helicase, that are essential for HCV proliferation. Therefore, NS3 is considered a target for anti-HCV treatment. To study RNA aptamers of the NS3 helicase domain, we carried out in vitro selection against the HCV NS3 helicase domain. RNA aptamers obtained after eight generations possessed 5' extended single-stranded regions and the conserved sequence (5'-GGA(U/C)GGAGCC-3') at stem-loop regions. Aptamer 5 showed strong inhibition of helicase activity in vitro. Deletion and mutagenesis analysis clarified that the conserved stem-loop is important and that the whole structure is needed for helicase inhibition. We compared the inhibition of helicase activity between aptamer 5 and 3'+-UTR of HCV.