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: 10245058
• 负责人: Louis M Mansky
• 金额: $ 19.38万
• 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 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10245058
• 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; Lentivirus; 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; 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 未成熟颗粒结构的关键分子接触，并告知 HIV- 1 蛋白水解裂解和 Gag 晶格拆卸。为了对逆转录病毒有一个基本的了解 需要正反转录病毒亚科的未成熟颗粒的组装和成熟、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）引导单粒子重建 （SPR），实现逆转录病毒未成熟颗粒的原子分辨率重建图。我们将利用人类 2 型免疫缺陷病毒 (HIV-2) 作为实施该图像处理策略的基础。我们 然后将该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），并对关键分子的保守性进行比较研究 HIV-1、HIV-2 和 HTLV-1 Gag 晶格结构中的相互作用。我们也将应用这种方法进行研究 膜锚定的 HIV-1 包膜蛋白结构。这种提出的混合图像处理方法 利用冷冻电镜成像方法（即冷冻 ET 和 SPR）的优点，并克服了 处理逆转录病毒未成熟颗粒图像时每种方法所面临的计算挑战。

项目成果

HIV-2 Immature Particle Morphology Provides Insights into Gag Lattice Stability and Virus Maturation.

HIV-2 未成熟颗粒形态学提供了对 Gag 晶格稳定性和病毒成熟的见解。

• 发表时间: 2023-08-01
• 影响因子: 5.6
• 作者: Talledge, Nathaniel;Yang, Huixin;Shi, Ke;Coray, Raffaele;Yu, Guichuan;Arndt, William G;Meng, Shuyu;Baxter, Gloria C;Mendonça, Luiza M;Castaño;Aihara, Hideki;Mansky, Louis M;Zhang, Wei
• 通讯作者: Zhang, Wei