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.

项目摘要/摘要 抗逆转录病毒靶标识别的一个关键方面是了解自然的分子相互作用，当 心烦意乱，会导致干扰人类免疫缺陷病毒1型(HIV-1)的复制。未被充分开发的 在HIV-1复制周期中发现新的抗病毒靶点的步骤一直是病毒 粒子集合。这在一定程度上是因为对病毒复制的这一阶段的理解相对较差-- 特别是因为它与Gag运动到质膜的行为有关，粒子的结合 萌芽位置，产生未成熟的Gag晶格的分子Gag-Gag相互作用，以及随后的粒子 生物发生学。对近亲的详细比较分析可以为发现关键 药物开发的抗逆转录病毒靶标。例如，最近的证据表明，恶作剧和恶作剧之间的互动 逆转录病毒之间的差异，这有助于解释未成熟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。 导致斑点形成以及粒子生物发生的本质也仍然知之甚少 逆转录病毒组装途径，特别是在人类免疫缺陷病毒1型及其近亲之间 --即人类免疫缺陷病毒2型(HIV-2)和人类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的结构和颗粒的生物发生 通过创新的最先进的实验方法。特别是，我们将应用低温电子 显微镜/断层摄影术(冷冻-EM/ET)、全内反射荧光显微镜(TIRF)，光激活 定位显微镜(Palm)和荧光涨落的单分子新技术 活细胞的光谱(FFS)研究1)未成熟的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本质，以及3)研究成核的途径 关于Gag齐聚的。这些新颖的研究利用创新技术，以提供新的见解。 进入艾滋病毒-1生命周期中一个非常重要且鲜为人知的方面，并导致确定 通过使用小分子干预来开发抗逆转录病毒靶标。