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分子相互作用，当 扰动，可能导致干扰人类免疫缺陷病毒1型（HIV-1）复制。一个不充分的 HIV-1复制周期用于发现新型抗病毒靶标的已成为病毒的步骤 粒子组件。这部分归因于对病毒复制阶段的相对不良理解 - 具体而言，它与插科打到质膜的行为有关，粒子的参与 萌芽的位点，产生未成熟og晶格的分子堵嘴相互作用，随后的粒子 生物发生。近亲的详细比较分析对于发现关键的比较可能是非常有用的 药物开发的抗逆转录病毒靶标。例如，最近的证据表明GAG-GAG相互作用 逆转录病毒之间的不同，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，我们已经对堵嘴核途径的差异进行了关键的初步观察 导致对刺的形成以及粒子生物发生的性质也仍然知之甚少 逆转录病毒组装途径，特别是在人类免疫缺陷病毒1型及其关闭中 亲戚 - 即2型人类免疫缺陷病毒（HIV-2）和人类T细胞白血病病毒1型（HTLV-1）。 在此应用中，我们建议研究逆转录病毒不成熟的颗粒结构和粒子生物发生 通过创新的最先进的实验方法。特别是，我们将应用冷冻电子 显微镜/断层扫描（冷冻EM/ET），总内反射荧光（TIRF）显微镜，光激活 定位显微镜（棕榈）和荧光波动的新型单分子技术 活细胞中的光谱法（FFS）研究1）未成熟og晶格结构的比较分析，2） 研究人逆转录病毒颗粒生物发生的性质，3）研究核的途径 GAG寡聚。这些新颖的研究利用创新技术来提供新的见解 成为HIV-1生命周期的高度重要且不理解的方面，并导致识别 抗逆转录病毒靶标，用于使用小分子进行干预。