Role of biomolecular condensates in regulating HIV-1 viral ribonucleoprotein complex formation in the setting of substance use disorder

摘要 HIV-1/艾滋病是一种毁灭性的免疫缺陷疾病，已导致3500多万人死亡 遍布地球仪。迫切需要开发新的治疗策略来对抗 抗药性病毒株的不断出现。一个尚未成功实现的药物靶点 临床是Gag蛋白与其RNA基因组相互作用形成病毒核糖核蛋白复合物， 其启动病毒体装配的过程。更深入地了解生物化学和生物物理 需要驱动病毒核糖核蛋白复合物形成的相互作用来推进进一步的治疗 发展基于最近的发现，病毒核糖核蛋白复合物经历液-液相 分离形成生物分子凝聚物，该提案探讨了GAG-病毒的分子基础 RNA相互作用我们已经组建了一个有成就的跨学科科学家团队， 逆转录病毒学，RNA生物学，生物物理学和药理学的交叉点，以阐明我们对 HIV-1感染中的病毒和细胞生物分子凝聚物。 我们的初步结果表明，HIV-1核糖核蛋白复合物在细胞核中形成， 这项研究病毒复制机制与核体相互作用的挑衅性提议， 形成生物分子凝聚物。在R21阶段，我们将使用涉及生物物理学的创新方法， 遗传学、最先进的活细胞成像和有针对性的药理学干预，以检查 HIV-1核糖核蛋白复合物在细胞核中组装成生物分子缩合物。在R33阶段， 我们将把这些研究扩展到探讨机制问题，重点是核生物导弹是否发挥关键作用， 在调节HIV-1转录、潜伏期再激活和基因组RNA包装中的作用。由于高 艾滋病毒1/艾滋病患者物质使用障碍的发病率，我们还将调查 滥用影响HIV-1核生物分子凝聚物。阐明HIV-1的遗传决定因素 冷凝物的形成可能导致治疗HIV/AIDS的新药物靶点的鉴定。

Abstract HIV-1/AIDS is a devastating immunodeficiency disease that has resulted in over 35 million deaths across the globe. There is a compelling ongoing need to develop novel treatment strategies to combat the continuing emergence of drug-resistant viral strains. A drug target that has not yet been successfully brought to the clinic is the interaction of the Gag protein with its RNA genome to form the viral ribonucleoprotein complex, which initiates the process of virion assembly. A deeper understanding of the biochemical and biophysical interactions that drive viral ribonucleoprotein complex formation is needed to advance further therapeutic development. Based on recent findings that viral ribonucleoprotein complexes undergo liquid-liquid phase separation to form biomolecular condensates, this proposal explores the molecular underpinnings of Gag-viral RNA interactions. We have assembled an accomplished interdisciplinary team of scientists to work at the crossroads of retrovirology, RNA biology, biophysics, and pharmacology to shed light on our understanding of viral and cellular biomolecular condensates in HIV-1 infection. Our preliminary results suggesting that HIV-1 ribonucleoprotein complexes form in the nucleus inspired this provocative proposal to investigate the interplay of virus replication machinery with nuclear bodies that form biomolecular condensates. In the R21 phase, we will use innovative methods involving biophysics, genetics, state-of-the-art live cell imaging, and targeted pharmacological interventions to examine whether HIV-1 ribonucleoprotein complexes assemble into biomolecular condensates in the nucleus. In the R33 phase, we will extend these studies to probe mechanistic questions focusing on whether nuclear BMCs play critical roles in regulating HIV-1 transcription, latency reactivation, and genomic RNA packaging. Due to the high incidence of substance use disorder in people with HIV-1/AIDS, we will also investigate whether drugs of abuse influence HIV-1 nuclear biomolecular condensates. Elucidating the genetic determinants of HIV-1 condensate formation could lead to the identification of novel drug targets to treat HIV/AIDS.