Cryo-ET Guided Single Particle Reconstruction of HIV

冷冻电子断层扫描引导的 HIV 单粒子重建

• 批准号: 10033396
• 负责人: Louis M Mansky
• 金额: $ 23.11万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-21 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10033396
• 关键词: 3-Dimensional; Alpha Particles; Binding; Cleaved cell; Comparative Study; Complex; Computing Methodologies; Crowding; Cryo-electron tomography; Cryoelectron Microscopy; Evaluation; Goals; HIV; HIV-1; HIV-2; Human T-lymphotropic virus 1; Hybrids; Image; Intervention; Ligands; Location; Maps; Membrane; Methods; Microscope; Molecular; Monoclonal Antibodies; Morphology; Positioning Attribute; Radiation induced damage; Research Personnel; Resolution; Retroviridae; Role; SP1 gene; Scheme; Specimen; Structure; Subfamily lentivirinae; Therapeutic Intervention; Tomogram; Ursidae Family; Viral; Virus; Virus Diseases; Virus-like particle; electron radiation; env Gene Products; image processing; imaging modality; particle; prevent; protein structure; reconstruction

ABSTRACT The atomic resolution structures of the human immunodeficiency virus type 1 (HIV-1) Gag lattice have revealed critical molecular contacts that stabilize the HIV-1 immature particle structure and have informed HIV- 1 proteolytic cleavages and Gag lattice disassembly. In order to gain a fundamental understanding of retrovirus assembly and maturation, atomic resolution maps of immature particles for the Orthoretrovirinae are required. Here we propose a strategy, i.e., cryo-electron tomography (cryo-ET) guided single particle reconstruction (SPR), to achieve atomic resolution reconstruction maps of retrovirus immature particles. We will use human immunodeficiency virus type 2 (HIV-2) as the basis for implementation of this image processing strategy. We will then apply the method to the study of other retrovirus immature particle structures, e.g., human T-cell leukemia virus type 1 (HTLV-1), and conduct comparative studies about the conservation of key molecular interactions in HIV-1, HIV-2 and HTLV-1 Gag lattice structures. We will also apply this approach for the study of membrane anchored HIV-1 envelope protein structure. This proposed hybrid image processing method takes advantage of the strengths of both cryo-EM imaging methods (i.e. cryo-ET and SPR) and overcomes the computation challenges that each method bears when processing images of retrovirus immature particles.

抽象的 人类免疫缺陷病毒1型（HIV-1）GAG晶格的原子分辨率结构具有 揭示了稳定HIV-1未成熟粒子结构的临界分子接触，并告知HIV- 1蛋白水解切割和插科打晶格拆卸。为了获得对逆转录病毒的基本了解 需要组装和成熟，原子分辨率图未成熟颗粒的原子分辨率图。 在这里，我们提出了一种策略，即冷冻电子断层扫描（Cryo-ET）指导的单粒子重建 （SPR），以实现逆转录病毒未成熟颗粒的原子分辨率重建图。我们将使用人类 免疫缺陷病毒2型（HIV-2）作为实施此图像处理策略的基础。我们 然后，将把方法应用于其他逆转录病毒未成熟颗粒结构的研究，例如人类T细胞 白血病病毒1型（HTLV-1），并进行有关关键分子保护的比较研究 HIV-1，HIV-2和HTLV-1 GAG晶格结构中的相互作用。我们还将将此方法应用于研究 膜锚定HIV-1包膜蛋白结构的结构。此提出的混合图像处理方法 利用两种低温EM成像方法（即冷冻-ET和SPR）的优势，并克服了 在处理逆转录病毒未成熟颗粒的图像时，每种方法都会挑战。