Center for Structural Biology of HIV RNA

摘要/摘要 在这个项目中，我们将分析HIV-1RNA分子的结构和分子转化 以基因组RNA(GRNA)的形式包装成病毒粒子。从细胞内的事件开始，我们将阐明如何 转录起始位点的使用定义了HIV-1RNA5‘-前导的结构，这反过来又决定了 该RNA将发生二聚化并作为gRNA发挥作用，或保持单体并作为信使RNA发挥作用。我们会 研究二聚体gRNA如何招募病毒Gag蛋白来产生一种复合体，从而使 未成熟的病毒粒子。最后，我们将研究病毒粒子成熟过程，在这个过程中，gRNA 浓缩成病毒核糖核蛋白复合体，重新包装在成熟的衣壳内，与 TRNA引物，并为反转录做准备。我们提议的研究将提供深刻的基础 对这些过程的理解，这些过程代表了一些最关键但不完全阐明的过程 HIV-1复制的分子机制。重要的是，我们现在也准备利用这一理解。 针对HIV-1RNA和HIV-1RNA的新型抑制剂的鉴定、表征和开发 RNA复合体。总体而言，我们的调查团队在确定 大RNA和RNA的结构：使用最先进的技术的蛋白质复合体；在分析它们的 在体外、在病毒粒子和在细胞中的生化和生物物理性质；在可视化这些复合体时 通过它们在细胞中的程序化过程和途径，以及在应用遗传学和病毒学 用于识别和表征宿主和病毒辅助因子、探测关键机制细节和测试小分子的工具 抑制剂。

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.