DNA aptamers selected against the HIV-1 RNase H display in vitro antiviral activity
DNA aptamers selected against the HIV-1 RNase H display in vitro antiviral activity
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DOI：
10.1021/bi0108599
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发表时间：
2001-08-28
期刊：
影响因子：
2.9
通讯作者：
Litvak, S
中科院分区：
文献类型：
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作者：
Andreola, ML;Pileur, F;Litvak, S
人类免疫缺陷病毒 1 型逆转录酶 (HIV-1 RT) 的 DNA 聚合酶是广泛用于抑制 HIV-1 复制的靶标。相比之下，尽管 RNase H 活性在病毒增殖中发挥着至关重要的作用，但与 RT 相关的 RNase H 活性抑制剂却很少被描述。使用具有或不具有 RNase H 结构域的重组 RT，通过指数富集策略 (SELEX) 进行配体系统进化，分离出对 HIV-1 RT RNase H 结构域具有高亲和力的 DNA 配体。所选寡核苷酸 (ODN) 能够在体外抑制 HIV-1 RNase H 活性，而对细胞 RNase H 没有观察到影响。我们将兴趣集中在两种富含 G 的抑制性寡核苷酸上。对这些 ODN 二级结构的模型研究强烈表明它们能够形成 G-四联体。除了在无细胞系统中观察到的 HIV-1 RNase H 抑制作用之外，这些 ODN 还能够强烈降低人类感染细胞中 HIV-1 的感染性。这里描述的寡核苷酸可以作为开发这种关键逆转录病毒酶活性的特异性抑制剂的主要化合物。
The DNA polymerase of the human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is a target widely used to inhibit HIV-1 replication. In contrast, very few inhibitors of the RNase H activity associated with RT have been described, despite the crucial role played by this activity in viral proliferation. DNA ligands with a high affinity for the RNase H domain of HIV-1 RT were isolated by systematic evolution of ligands by an exponential enrichment strategy (SELEX), using recombinant RTs with or without the RNase H domain. The selected oligonucleotides (ODNs) were able to inhibit in vitro the HIV-1 RNase H activity, while no effect was observed on cellular RNase H. We focused our interest on two G-rich inhibitory oligonucleotides. Model studies of the secondary structure of these ODNs strongly suggested that they were able to form G-quartets. In addition to the inhibition of HIV-1 RNase H observed in a cell free system, these ODNs were able to strongly diminish the infectivity of HIV-1 in human infected cells. Oligonucleotides described here may serve as leading compounds for the development of specific inhibitors of this key retroviral enzyme activity.