Center for Structural Biology of HIV RNA

总结/摘要 在本项目中，我们将分析HIV-1 RNA分子的结构和分子转化， 作为基因组RNA（gRNA）被包装到病毒体中。从细胞中的事件开始，我们将阐明 转录起始位点的使用定义了HIV-1 RNA 5 '前导序列的结构，反过来，它决定了是否 RNA将二聚化并作为gRNA起作用，或保持单体并作为信使RNA起作用。我们将 研究二聚体gRNA如何招募病毒Gag蛋白以产生复合物，该复合物使病毒的组装成核。 未成熟病毒体。最后，我们将研究病毒体成熟过程中发生的过程，其中gRNA 浓缩成病毒核糖核蛋白复合物，在成熟衣壳内重新包装， tRNA引物，并准备反转录。我们提出的研究将提供深刻的基础 理解这些过程，这代表了一些最关键的，但不完全阐明 HIV-1复制的分子机制。重要的是，我们现在也准备利用这种理解 致力于鉴定、表征和开发针对HIV-1 RNA的新型抑制剂， RNA复合物。总的来说，我们的调查团队在确定 大RNA和RNA的结构：蛋白质复合物使用国家的最先进的技术;在分析他们的 在体外，在病毒体和细胞中的生物化学和生物物理特性;在观察复合物时， 在细胞中通过其程序化的过程和途径进行;并在应用遗传学和病毒学 用于识别和表征宿主和病毒辅因子、探测关键机制细节和测试小分子的工具 抑制剂的

Summary/Abstract In this Project, we will analyze the structures and molecular transformations of the HIV-1 RNA molecules that become packaged into virions as genomic RNA (gRNA). Beginning with events in the cell, we will elucidate how transcription start site usage defines the structure of the HIV-1 RNA 5’-leader, which, in turn, dictates whether the RNA will dimerize and function as gRNA or remain monomeric and function as messenger RNA. We will study how the dimeric gRNA recruits the viral Gag protein to generate a complex that nucleates assembly of the immature virion. Finally, we will look at processes that occur during virion maturation, in which the gRNA condenses into a viral ribonucleoprotein complex, is re-packaged inside the mature capsid, anneals with the tRNA primer, and prepares for reverse transcription. Our proposed studies will provide deep fundamental understanding of these processes, which represent some of the most critical but incompletely elucidated molecular mechanisms in HIV-1 replication. Importantly, we are now also poised to exploit this understanding towards the identification, characterization and development of novel inhibitors targeted against HIV-1 RNA and RNA complexes. Collectively, our team of investigators have a strong track record and expertise in determining structures of large RNAs and RNA:protein complexes using state-of-the-art techniques; in analyzing their biochemical and biophysical properties in vitro, in virions, and in cells; in visualizing the complexes as they proceed through their programmed processes and pathways in the cell; and in applying genetic and virological tools to identify and characterize host and viral co-factors, probe key mechanistic details, and test small molecule inhibitors.