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

项目摘要 人类免疫缺陷病毒1型(HIV-1)已经进化出复杂的策略来限制呈现 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的数量就证明了这一点。 暴露在病毒表面的包膜蛋白，以牺牲病毒的传染性为代价。HIV-1是如何调节 病毒包膜蛋白在组装颗粒中的掺入仍然非常不清楚。我们假设 HIV-1囊膜胞浆结构域和宿主细胞膜脂质中发现的特定基序 规范环境病毒的获取，使其组装成病毒颗粒。在我们这项提案的第一个目标中，我们将量化 HIV-1囊膜及其胞浆结构域突变体的单分子纳米动力学研究 生产细胞的表面。在第二个目标中，我们将测量三维(3D)结构 萌发病毒颗粒表面HIV-1包膜的组织和角分布对两个人都是 目的，我们建议确定关键的病毒和宿主细胞膜决定因素负责时空 HIV-1包膜与新生病毒组装部位的关联。这两个目标都将利用数十纳米 三维超分辨显微镜解释动力学的分辨率和分子特异性 和HIV-1外膜的组织。我们的量化方法将提供必要的基础 关于构建HIV-1病毒包膜获取到病毒组装部位的准确模型的知识。我们 相信这些模型将有助于未来开发旨在抑制可获得性的新战略 HIV-1包膜到病毒组装部位。

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.