Ojha Manju, Hudson Lucia, Photenhauer Amanda, Zang Trinity, Lerew Lauren, Ekesan Şölen, Daniels Jason, Nguyen Megan, Paudyal Hardik, York Darrin M, Ohi Melanie D, Marchant Jan, Bieniasz Paul D, Koirala Deepak
Department of Chemistry and Biochemistry, University of Maryland Baltimore County; Baltimore, MD, USA.
Life Sciences Institute, University of Michigan; Ann Arbor, MI, USA.
bioRxiv. 2025 Feb 26:2025.02.20.639096. doi: 10.1101/2025.02.20.639096.
The Rev Response Element (RRE) forms an oligomeric complex with the viral protein Rev to facilitate the nuclear export of intron-retaining viral RNAs during the late phase of HIV-1 infection. However, our structural understanding of this crucial virological process remains limited. In this study, we determined several crystal structures of an intact RRE stem-loop II in two distinct conformations, performed negative-staining electron microscopy and molecular dynamics simulations, and revealed that this three-way junction RNA exhibits remarkable structural plasticity. Through Rev-binding and Rev-activity assays using various stem-loop II mutants designed to favor one of the conformers, we demonstrated that the structural plasticity of stem-loop II modulates Rev binding and oligomerization. Our findings illuminate emerging perspectives on RRE dynamics-based regulation of HIV-1 RNA nuclear export and provide a framework for developing anti-HIV drugs that target specific RRE conformations.
Rev反应元件（RRE）与病毒蛋白Rev形成寡聚复合物，以促进HIV-1感染后期保留内含子的病毒RNA的核输出。然而，我们对这一关键病毒学过程的结构理解仍然有限。在本研究中，我们确定了完整的RRE茎环II的几种处于两种不同构象的晶体结构，进行了负染色电子显微镜和分子动力学模拟，并揭示这种三向接头RNA具有显著的结构可塑性。通过使用设计为有利于其中一种构象体的各种茎环II突变体进行Rev结合和Rev活性测定，我们证明茎环II的结构可塑性调节Rev结合和寡聚化。我们的研究结果阐明了基于RRE动力学的HIV-1 RNA核输出调节的新观点，并为开发靶向特定RRE构象的抗HIV药物提供了框架。
