Cryo-Electron Tomography Analysis of HIV-1 Maturation

我们建议通过以下方式确定在选定的成熟阶段停止的HIV-1 VLP的结构 冷冻电子断层扫描。这种方法将使我们能够确定(1)病毒的形态变化 在成熟过程中经历，(2)未成熟和成熟的HIV-1病毒囊膜蛋白的结构 表达的蛋白(Env)及其在基质(MA)层中的排列方式，以及(3)其他病毒的位置 粒子内的组件。为了实现这些目标，与该大学的研究人员合作 在犹他州，我们计划(1)提纯在几个发育阶段停止的HIV-1变异株，(2)想象HIV-1 通过低温电子断层扫描和通过计算方法计算结构，以及(3) 确定特定病毒蛋白在成熟过程中存在的结构变异性。我们的研究将 提供完整的、完整的冷冻水合HIV-1 VLP的最高分辨率结构 研究人员可能会确定其他蛋白质的位置和可能产生新疗法的数据 旨在抗击或根除艾滋病毒-1感染。

We propose to determine the structure of HIV-1 VLPs halted at selected stages throughout maturation by cryo-electron tomography. This method will enable us to determine (1) the morphological changes the virus undergoes during maturation, (2) the structure of immature and mature HIV-1 VLPs with the Envelope protein (Env) expressed and how it is arranged in the matrix (MA) layer, and (3) the positions of other viral components within the particles. To achieve these aims, in collaboration with researchers at the University of Utah, we plan to (1) purify HIV-1 mutants halted at several 'developmental' stages, (2) image the HIV-1 mutants by cryo-electron tomography and calculate the structures through computational methods, and (3) determine the structural variability present in the specific viral proteins during maturation. Our studies will provide the highest resolution structures of whole, intact frozen-hydrated HIV-1 VLPs from which other researchers may determine the placement of other proteins and data from which new therapies may be designed to combat or eradicate HIV-1 infection.