Korolev Sergey, Knyazhanskaya Ekaterina, Anisenko Andrey, Tashlitskii Vadim, Zatsepin Timofei S, Gottikh Marina, Agapkina Julia
Department of Chemistry, Belozersky Institute of Physical and Chemical Biology, Moscow State University, Moscow, Russia.
Nucleosides Nucleotides Nucleic Acids. 2011 Jul-Aug;30(7-8):651-66. doi: 10.1080/15257770.2011.592890.
Integration of the DNA copy of the genomic RNA into an infected cell genome is one of the key steps of the replication cycle of all retroviruses. It is catalyzed by the viral enzyme, integrase. We have shown that conjugates of short single-stranded oligonucleotides with eosin efficiently inhibit the catalytic activity of the HIV-1 integrase. In this article, we have found that the dependence of the integrase catalytic activity on the concentration of oligonucleotides has a bell-shaped pattern. The modulation of HIV-1 integrase activity correlated with the oligonucleotide length and was not associated with specific sequences. Moreover, a similar mode of the oligonucleotide action was found for integrase from the prototype foamy virus. This dual effect of the oligonucleotide and their conjugates with eosin might be explained by their binding with retroviral integrase in two different sites; the oligodeoxynucleotide binding in the first site results in integrase activation, whereas interactions with another one lead to inhibition of the enzyme activity. Eosin coupling to oligonucleotides did not change the mode of their action but enhanced their affinity to both binding sites. The affinity increase was found to be much more important for the site responsible for the integrase inhibition, thus explaining the high inhibitory potency of oligonucleotide-eosin conjugates.
基因组RNA的DNA拷贝整合到受感染细胞基因组中是所有逆转录病毒复制周期的关键步骤之一。它由病毒酶整合酶催化。我们已经表明，短单链寡核苷酸与曙红的缀合物能有效抑制HIV-1整合酶的催化活性。在本文中，我们发现整合酶催化活性对寡核苷酸浓度的依赖性呈钟形模式。HIV-1整合酶活性的调节与寡核苷酸长度相关，且与特定序列无关。此外，对于原型泡沫病毒的整合酶，也发现了类似的寡核苷酸作用模式。寡核苷酸及其与曙红的缀合物的这种双重作用可能是由于它们在两个不同位点与逆转录病毒整合酶结合；寡脱氧核苷酸在第一个位点的结合导致整合酶激活，而与另一个位点的相互作用则导致酶活性受到抑制。曙红与寡核苷酸的偶联并没有改变它们的作用方式，但增强了它们对两个结合位点的亲和力。对于负责整合酶抑制的位点，亲和力的增加更为重要，这就解释了寡核苷酸-曙红缀合物的高抑制效力。
