HIV Gag lattice morphology and particle biogenesis - Administrative Supplement

HIV Gag 晶格形态和颗粒生物发生 - 行政补充

• 批准号: 9701541
• 负责人: JOACHIM D MUELLER
• 金额: $ 17.04万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9701541
• 关键词: Administrative Supplement; Anti-HIV Therapy; Anti-Retroviral Agents; Antiviral Agents; Behavior; Binding; Biogenesis; Cell membrane; Cells; Cryoelectron Microscopy; Development; Drug Targeting; Fluorescence; Fluorescence Microscopy; HIV; HIV Infections; HIV-1; HIV-2; Human Characteristics; Human T-lymphotropic virus 1; Intervention; Knowledge; Lead; Life Cycle Stages; Microscopy; Molecular; Morphology; Movement; Nature; Pathway interactions; Pharmaceutical Preparations; Phase; Retroviridae; Site; Spectrum Analysis; Structure; Technology; Virion; Virus Replication; base; comparative; drug development; innovation; innovative technologies; insight; novel; particle; single molecule; small molecule; tomography

Project Summary/Abstract A key aspect to antiretroviral target identification is understanding the nature molecular interactions, which when perturbed, can result interfere with human immunodeficiency virus type 1 (HIV-1) replication. An underexplored aspect of the HIV-1 replication cycle for discovery of novel antiviral targets has been the steps involved in virus particle assembly. This is due, in part, to the relatively poor understanding of this phase of virus replication – specifically as it relates to the behavior of Gag movement to the plasma membrane, the engagement of particle budding sites, the molecular Gag-Gag interactions that create the immature Gag lattice, and subsequent particle biogenesis. Detailed comparative analysis of close relatives can be highly informative for the discovery of key antiretroviral targets for drug development. For instance, recent evidence indicates that Gag-Gag interactions differ among retroviruses, which helps explain morphological differences among immature retrovirus particles. Furthermore, we have made key preliminary observations of differences in the pathways for Gag nucleation leading to punctum formation, as well as the nature of particle biogenesis also remain poorly understood aspects of the retrovirus assembly pathway, particularly among human immunodeficiency virus type 1 and its close relatives – i.e., human immunodeficiency virus type 2 (HIV-2) and human T-cell leukemia virus type 1 (HTLV-1). In this application, we propose to investigate retrovirus immature particle structure and particle biogenesis through innovative state-of-the-art experimental approaches. In particular, we will apply cryo-electron microscopy/tomography (cryo-EM/ET), total internal reflection fluorescence (TIRF) microscopy, photoactivated localization microscopy (PALM), and the novel single-molecule technology of fluorescence fluctuation spectroscopy (FFS) in living cells to investigate 1) comparative analysis of immature Gag lattice structures, 2) investigate the nature of human retrovirus particle biogenesis, and 3) investigate the pathways for the nucleation of Gag oligomerization. These novel studies harness innovative technologies in order to provide new insights into a highly significant and poorly understood aspect of the HIV-1 life cycle and lead to the identification of antiretroviral targets for exploiting by intervention using small molecules.

项目总结/摘要 抗逆转录病毒靶点鉴定的一个关键方面是了解分子相互作用的性质， 干扰，可导致干扰人类免疫缺陷病毒1型（HIV-1）的复制。一个探索不足的 用于发现新的抗病毒靶点的HIV-1复制周期的一个方面是涉及病毒复制的步骤。 粒子组装这部分是由于对病毒复制的这一阶段的了解相对较少- 特别是因为它涉及Gag运动到质膜的行为，颗粒的接合， 出芽位点，产生未成熟Gag晶格的分子Gag-Gag相互作用，以及随后的粒子 生物起源对近亲的详细比较分析可以为发现关键基因提供大量信息。 抗逆转录病毒药物开发的目标。例如，最近的证据表明， 不同的逆转录病毒，这有助于解释不成熟的逆转录病毒颗粒之间的形态差异。 此外，我们还对Gag成核途径的差异进行了关键的初步观察 导致泪点形成以及颗粒生物发生的本质仍然是人们知之甚少的方面 逆转录病毒装配途径，特别是在人类免疫缺陷病毒1型及其密切 亲属-即，人免疫缺陷病毒2型（HIV-2）和人T细胞白血病病毒1型（HTLV-1）。 在这个应用中，我们建议研究逆转录病毒未成熟颗粒结构和颗粒生物发生 通过最先进的实验方法。特别是，我们将应用低温电子 显微镜/断层扫描（cryo-EM/ET），全内反射荧光（TIRF）显微镜，光活化 定位显微镜（PALM）和荧光波动的新的单分子技术 在活细胞中使用FFS来研究1）未成熟Gag晶格结构的比较分析，2） 研究人类逆转录病毒颗粒生物发生的性质，以及3）研究成核的途径 Gag寡聚化。这些新的研究利用创新技术，以提供新的见解 艾滋病毒-1生命周期的一个非常重要和知之甚少的方面，并导致识别 抗逆转录病毒靶点，通过使用小分子进行干预来利用。