Project 2 - Nuclear Export and Translation

项目2：核出口和翻译 摘要 HIV-1 RNA从细胞核转运及其在细胞质中的调节表达是 病毒的生命周期。病毒RNA中存在的结构域已被证明调节这些事件；即，输出 非剪接转录本需要REV反应元件，Rre；5ʹ-Leader的替代构象 区域决定了翻译与包装的命运；以及对遗传信息的重新编码，使得 固定比例的病毒蛋白的产生依赖于移码信号。在阐明这些RNA的同时 在CRNA中进行的研究表明，结构以及调节这些事件的机制更接近于实现 1.0已经确定，核出口和翻译监管都比 之前的想法是。有几个重大发现是以前没有预测或预见到的：细胞质 GAG：RRE相互作用；转录起始点异质性的存在及其对 维护由5个ʹ-Leader结构组成的单独池；以及在 不活跃的茎环和活跃的、允许移码的假结构象。基于这些CRNA 发现，这个项目的目的是获得一个完整的结构和机制上的理解 结构研究与可视化、生化相结合的HIV-1转运和翻译调控 和病毒学实验。我们理解核出口的目的将是确定高分辨率 RRE核输出信号的结构，GAG：RRE相互作用的生物学意义，以及 各种HIV-1核糖核酸的核出口特性。我们理解翻译法规的目标将是 确定5个ʹ-Leader的单体和剪接env mrna形式的结构，以及 参与了程序性核糖体移码的过程。 好了！

Project 2: Nuclear export and translation Summary Transport of HIV-1 RNAs from the nucleus and their regulated expression in the cytoplasm are critical steps of the viral lifecycle. Domains present in viral RNA have been shown to regulate these events; namely, the export of unspliced transcripts requires the Rev responsive element, RRE; alternative conformers of the 5ʹ-leader region dictate translation vs. packaging fates; and the recoding of genetic information that allows the production of fixed ratios of viral proteins relies on the frameshifting signal. While elucidation of these RNA structures, and thus mechanisms regulating these events, are closer to realization, studies carried out in CRNA 1.0 have established that both nuclear export and translational regulation are more multifaceted than previously thought. Several major discoveries were not previously predicted or envisioned: the cytoplasmic Gag:RRE interaction; the presence of transcriptional start site heterogeneity, and its consequence on maintaining separate pools of 5ʹ-leader structures; and the presence of a structural equilibrium between an inactive stem-loop and an active, frameshift-permissive pseudoknot conformation. Based on these CRNA discoveries, this project aims to gain a complete structural and mechanistic understanding of both nuclear transport and translational regulation in HIV-1 by combining structural studies with visualization, biochemical and virologic experiments. Our aims for understanding nuclear export will be to determine the high resolution structures of the RRE nuclear export signal, the biological significance of Gag:RRE interactions, and the nuclear export properties of various HIV-1 RNAs. Our aims for understanding translational regulation will be to determine structures for the monomeric and the spliced env mRNA forms of the 5ʹ-leader, and the structures involved in the process of programmed ribosomal frameshifting. !