Elucidation and characterization of oligonucleotide-accessible sites on HIV-2 leader region RNA.
Elucidation and characterization of oligonucleotide-accessible sites on HIV-2 leader region RNA.
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HIV-2 前导区 RNA 上寡核苷酸可及位点的阐明和表征。
DOI：
10.1089/108729003764097331
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发表时间：
2003
期刊：
影响因子：
--
通讯作者：
Lodmell,JStephen
中科院分区：
文献类型：
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作者：
Deer,EmilyL;Douk,Boramee;Lanchy,Jean-Marc;Lodmell,JStephen
逆转录病毒，包括人类病原体HIV-1和HIV-2，是二倍体，因为它们使其RNA基因组的两个拷贝双化。在双核苷化之前或期间，两个拷贝的全长基因组 RNA在一个称为二聚化的过程中相互识别并稳定结合。病毒基因组内的RNA结构促进HIV-1和HIV-2的二聚化，并位于5′-非翻译区。 leader区域通过这些RNA信号的突变来抑制二聚化已被证明可以显著降低病毒感染性和复制动力学，因此，代表了潜在的靶点 进行抗逆转录病毒治疗在这项研究中，我们确定了HIV-2前导区RNA中的位点，这些位点在功能上可以通过随机逆转录与寡核苷酸（ODNs）杂交， ODN库（RT-ROL）。然后，我们测试了针对这些区域的特异性ODNs在体外抑制RNA二聚化的功效。我们确定了几个杂交能力强的 ODNs，只有两个是非常有效的抑制RNA二聚化。这两种ODN在引物结合位点（PBS）与病毒RNA互补。这些结果确定了具有高可达性的区域 与HIV-2 RNA上的ODN结合，并有助于绘制病毒RNA内二聚化所必需的区域。
The retroviruses, including the human pathogens HIV-1 and HIV-2, are diploid inasmuch as they encapsidate two copies of their RNA genome. Prior to or during encapsidation, two copies of full-length genomic RNA recognize and stably bind each other in a process called dimerization. RNA structures within the viral genome promote dimerization in both HIV-1 and HIV-2 and are located in the 5′-untranslated leader region. Inhibition of dimerization by mutation of these RNA signals has been demonstrated to drastically reduce viral infectivity and replication kinetics and, thus, represents a potential target for antiretroviral therapy. In this study, we identified sites in HIV-2 leader region RNA that are functionally accessible to hybridization with oligonucleotides (ODNs) by reverse transcription with random ODN libraries (RT-ROL). We then tested specific ODNs directed against these regions for their efficacy in inhibiting RNA dimerizationin vitro. We determined that of several hybridization-competent ODNs, only two were very effective in inhibiting RNA dimerization. Both of these ODNs were complementary to viral RNA at the primer binding site (PBS). These results identify regions with high accessibility to ODN binding on HIV-2 RNA and help to map the region(s) essential for dimerization within the viral RNA.