Defining the three-dimensional organization and dynamics of HIV-1 Envelope using superresolution microscopy

使用超分辨率显微镜定义 HIV-1 包膜的三维组织和动态

• 批准号: 9203213
• 负责人: Schuyler van Engelenburg
• 金额: $ 20.85万
• 依托单位: UNIVERSITY OF DENVER (COLORADO SEMINARY)
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-16 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9203213
• 关键词: Biogenesis; Biological; Cell membrane; Cells; Complex; Cytoplasmic Tail; Data; Data Set; Development; Diffusion; Elements; Epitopes; Future; Glycoproteins; Goals; HIV-1; Image; Immune system; Infection; Knowledge; Label; Life; Lipids; Measurement; Measures; Mediating; Membrane; Membrane Lipids; Methodology; Microscopy; Modeling; Molecular; Monitor; Mutation; Process; Protein Dynamics; Protocols documentation; Recruitment Activity; Resolution; Site; Spatial Distribution; Specificity; Staging; Structure; Surface; System; Techniques; Testing; Viral; Viral Envelope Proteins; Viral Proteins; Virion; Virus; Virus Assembly; Virus Diseases; cellular targeting; combat; env Gene Products; gag Gene Products; insight; mutant; nanometer; nanoscale; novel strategies; novel therapeutic intervention; operation; particle; prevent; single molecule; spatiotemporal; targeted treatment

Project Summary Human Immunodeficiency Virus 1 (HIV-1) has evolved sophisticated strategies to limit the presentation of viral epitopes to the immune system. This is evidenced by the ability of the virus to minimize the number of exposed Envelope proteins on the virus surface at the expense of virus infectivity. How HIV-1 regulates the incorporation of viral Envelope proteins into assembling particles is still very unclear. We hypothesize that specific motifs found within the HIV-1 Envelope cytoplasmic domain as well as host cell membrane lipids regulate the acquisition of Env into assembling virus particles. In our first aim of this proposal, we will quantify the single molecule nanoscale dynamics of HIV-1 Envelope and mutants of the cytoplasmic domain on the surface of producing cells. In our second aim, we will measure the three-dimensional (3D) structural organization and angular distribution of HIV-1 Envelope on the surface of budding virus particles. For both aims, we propose to identify key viral and host cell membrane determinants responsible for the spatiotemporal association of HIV-1 Envelope with nascent virus assembly sites. Both aims will utilize the tens of nanometer resolution and molecular specificity of three-dimensional superresolution microscopy to illuminate the dynamics and organization of HIV-1 Envelope. Our quantitative approach will provide the necessary foundational knowledge for constructing accurate models of HIV-1 Envelope acquisition into virus assembly sites. We believe that these models will aid in the future development of novel strategies aiming to inhibit accessibility of HIV-1 Envelope to virus assembly sites.

项目概要 人类免疫缺陷病毒 1 (HIV-1) 已进化出复杂的策略来限制其表现 病毒表位对免疫系统的影响。病毒能够最大限度地减少病毒数量就证明了这一点。 以牺牲病毒感染性为代价，将包膜蛋白暴露在病毒表面。 HIV-1如何调节 将病毒包膜蛋白掺入组装颗粒中仍然非常不清楚。我们假设 HIV-1 包膜胞质结构域以及宿主细胞膜脂质中发现的特定基序 调节 Env 的获取以组装病毒颗粒。在本提案的第一个目标中，我们将量化 HIV-1包膜和胞质结构域突变体的单分子纳米级动力学 生产细胞的表面。在我们的第二个目标中，我们将测量三维 (3D) 结构 出芽病毒颗粒表面 HIV-1 包膜的组织和角度分布。对于两者 目标，我们建议确定负责时空的关键病毒和宿主细胞膜决定因素 HIV-1 包膜与新生病毒组装位点的关联。这两个目标都将利用数十纳米 三维超分辨率显微镜的分辨率和分子特异性来阐明动力学 HIV-1 包膜的组织和组织。我们的定量方法将提供必要的基础 构建将 HIV-1 包膜采集到病毒组装位点的准确模型的知识。我们 相信这些模型将有助于未来开发旨在抑制可访问性的新策略 HIV-1 病毒组装位点的包膜。