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

摘要 HIV-1/AIDS是一种毁灭性的免疫缺陷疾病，已导致3500万人死亡 在全球各地。迫切需要开发新的治疗策略来对抗 抗药性病毒株不断出现。尚未成功地将毒品目标带到 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体， 这启动了病毒粒子组装的过程。加深对生物化学和生物物理学的理解 推动病毒核糖核蛋白复合体形成的相互作用需要进一步的治疗 发展。基于最近的发现，病毒核糖核蛋白复合体经历液-液相 分离形成生物分子凝聚体，这一提议探索了Gag-Virus的分子基础 RNA相互作用。我们已经组建了一支经验丰富的跨学科科学家团队，在 逆转录病毒学、RNA生物学、生物物理学和药理学的十字路口，以阐明我们对 HIV-1感染中的病毒和细胞生物分子凝聚。 我们的初步结果表明，HIV-1核糖核蛋白复合体是在由 这一具有挑衅性的建议旨在调查病毒复制机制与核机构的相互作用 形成生物分子凝聚体。在R21阶段，我们将使用涉及生物物理的创新方法， 遗传学，最先进的活细胞成像，以及有针对性的药物干预，以检查 HIV-1核糖核蛋白复合体在细胞核内组装成生物分子凝聚体。在R33阶段， 我们将扩展这些研究，以探索机制问题，重点是核BMC是否发挥关键作用 在调节HIV-1转录、潜伏期重新激活和基因组RNA包装中的作用。由于高企 HIV-1/AIDS患者中药物使用障碍的发生率，我们还将调查药物是否 滥用会影响HIV-1核生物分子凝聚体。HIV-1基因决定因素的阐明 凝析油的形成可能导致确定治疗艾滋病毒/艾滋病的新药物靶标。

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.