Andreola M L, Pileur F, Calmels C, Ventura M, Tarrago-Litvak L, Toulmé J J, Litvak S
UMR 5097 CNRS-Université Victor Segalen Bordeaux 2, 146, rue Léo Saignat, 33076 Bordeaux Cedex, France.
Biochemistry. 2001 Aug 28;40(34):10087-94. doi: 10.1021/bi0108599.
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.
人类免疫缺陷病毒1型逆转录酶（HIV-1 RT）的DNA聚合酶是一种广泛用于抑制HIV-1复制的靶点。相比之下，尽管与RT相关的RNase H活性在病毒增殖中起着关键作用，但描述的针对该活性的抑制剂却很少。通过指数富集配体系统进化策略（SELEX），使用带有或不带有RNase H结构域的重组RT，分离出了对HIV-1 RT的RNase H结构域具有高亲和力的DNA配体。所选的寡核苷酸（ODN）能够在体外抑制HIV-1的RNase H活性，而对细胞RNase H没有影响。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对HIV-1 RNase H的抑制作用外，这些ODN还能够显著降低HIV-1在人类感染细胞中的感染性。本文所述的寡核苷酸可作为开发这种关键逆转录病毒酶活性特异性抑制剂的先导化合物。
