Project 4 - RNA interactions during HIV-1 assembly and maturation

摘要 虽然Gag-5的前导相互作用对于将HIV-1基因组整合到病毒粒子中是至关重要的，但它绝不是 意味着形成形态准确、具有感染性的唯一RNA蛋白质相互作用 病毒粒子。事实上，CRNA研究人员已经发现了HIV-1蛋白之间的非规范相互作用 (基质、整合酶、蛋白酶)以及宿主和/或病毒来源的RNA分子，这些分子对 规范传染性病毒粒子形成的关键事件的时间和地点。此外，CRNA 研究人员在确定RNA：RNA相互作用(tRNA：U5-PBS)的结构方面取得了关键进展 这也是在病毒粒子形态形成过程中启动的，对于感染性病毒粒子的产生同样至关重要。 项目4将由结构和功能实验组成，旨在阐明 这些事件的分子细节及其在HIV-1病毒粒子发生过程中的重要性。在《目标1：夏天》中，张 Bieniasz将确定基质的结构：似乎调节Gag：膜的tRNA复合体 结合，在目标2 D‘Souza，Smith和Chiu将决定tRNA：U5-PBS：反向的结构和功能 在病毒粒子成熟过程中形成的转录酶复合体，但对启动随后的 逆转录。在目标3中，Kutluay和Smith将确定IN：RNA复合体的结构 似乎对核糖核蛋白复合体在成虫衣壳核心内的正确定位至关重要 病毒粒子，并将确定抑制这种与小分子相互作用的可行性。最后，在目标4中 Swanstrom和Bieniasz将确定在体外激活蛋白酶的RNA：蛋白酶相互作用是否也 病毒粒子以及它们的功能意义，以期最终确定 这种互动的特点。

Summary While the Gag-5' leader interaction is crucial for the incorporation of HIV-1 genomes into virions, it is by no means the only RNA protein interaction that is required for the formation of morphologically accurate, infectious virions. Indeed, CRNA investigators have discovered non-canonical interactions between HIV-1 proteins (matrix, integrase, protease) and RNA molecules of host and/or viral origin that are, or may be, crucial for regulating the timing and location of key events in the formation of infectious virions. Additionally, CRNA investigators have made key advances in determining the structure of an RNA:RNA interaction (tRNA:U5-PBS) that is also initiated during virion morphogenesis and is similarly crucial for the generation of infectious virions. Project 4 will consist of a mixture of structural and functional experiments that will aim to elucidate the molecular details of these events and their importance during HIV-1 virion genesis. In Aim 1 Summers, Zhang and Bieniasz will determine the structures of matrix:tRNA complexes that appear to regulate Gag:membrane binding, in Aim 2 D’Souza, Smith, and Chiu will determine structures and functions of tRNA:U5-PBS:reverse transcriptase complexes that are formed during virion maturation, but are crucial for initiating subsequent reverse transcription. In Aim 3, Kutluay and Smith will determine the structure of IN:RNA complexes that appear crucial for the correct positioning of the ribonucleoprotein complex within the capsid core of mature virions, and will determine the feasibility of inhibiting this interaction with small molecules. Finally, in Aim 4 Swanstrom and Bieniasz will determine whether RNA:protease interactions that activate protease in vitro also occur in virions, as well as their functional significance, with a view to eventually determining the structural features of this interaction.