HIV-1 Packaging Visualised by In-Gel SHAPE.

HIV-1 Packaging Visualised by In-Gel SHAPE.
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DOI:
10.3390/v13122389
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发表时间:
2021-11-29
期刊:
Viruses
影响因子:
--
通讯作者:
Kenyon JC
Kenyon JC
中科院分区:
其他
文献类型:
--
作者:
D'Souza AR;Jayaraman D;Long Z;Zeng J;Prestwood LJ;Chan C;Kappei D;Lever AML;Kenyon JC

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HIV-1通过与病毒结构蛋白Gag的相互作用将其gRNA的两个拷贝包装成病毒体。这两个拷贝和它们的天然RNA结构对于病毒体的感染性是必不可少的。包装过程的精确逐步性质尚未解决。这在很大程度上是由于之前缺乏跟踪RNA-蛋白质复合物内RNA结构变化的结构技术。在这里,我们应用凝胶内SHAPE(引物延伸分析的选择性2 'OH酰化)技术来研究HIV-1包装的起始,检查包装信号RNA和Gag多蛋白之间的相互作用,并将其与单独的Gag NC结构域进行比较。我们的研究结果意味着在切换到一个二聚化能力的结构的RNA构象的Gag和单体包装信号RNA之间的相互作用,并显示,然后继续稳定的Gag-二聚体复合物。这些数据为HIV-1如何调节其基因组的翻译和包装提供了新的见解。
HIV-1 packages two copies of its gRNA into virions via an interaction with the viral structural protein Gag. Both copies and their native RNA structure are essential for virion infectivity. The precise stepwise nature of the packaging process has not been resolved. This is largely due to a prior lack of structural techniques that follow RNA structural changes within an RNA–protein complex. Here, we apply the in-gel SHAPE (selective 2’OH acylation analysed by primer extension) technique to study the initiation of HIV-1 packaging, examining the interaction between the packaging signal RNA and the Gag polyprotein, and compare it with that of the NC domain of Gag alone. Our results imply interactions between Gag and monomeric packaging signal RNA in switching the RNA conformation into a dimerisation-competent structure, and show that the Gag–dimer complex then continues to stabilise. These data provide a novel insight into how HIV-1 regulates the translation and packaging of its genome.
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