HIV particle morphology and biogenesis

HIV颗粒形态和生物发生

• 批准号: 10772748
• 负责人: JOACHIM D MUELLER
• 金额: $ 71.21万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-07 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10772748
• 关键词: 3-Dimensional; Acquired Immunodeficiency Syndrome; Actins; Address; Anti-Retroviral Agents; Appearance; Basic Science; Behavior; Binding; Biogenesis; Biophysics; Capsid; Capsid Proteins; Cell membrane; Cells; Color; Complex; Cryoelectron Microscopy; Cytoplasm; Development; Environment; Event; Exclusion; Fluorescence; Fluorescence Microscopy; HIV; HIV therapy; HIV-1; HIV-2; Human; Image; Infection; Integrase; Investigation; Knowledge; Lentivirus; Life Cycle Stages; Measurement; Membrane; Microscopy; Modeling; Morphogenesis; Morphology; Nature; Pathway interactions; Peptide Hydrolases; Process; Research; Resolution; Retroviridae; Retrovirology; Reverse Transcriptase Inhibitors; Ribonucleoproteins; Role; SP1 gene; Scanning; Site; Stress; Structure; Techniques; Technology; Therapeutic Intervention; Thick; Viral; Virion; Virus Assembly; antiretroviral therapy; cell type; comparative; experimental study; flexibility; fluorescence imaging; gag Gene Products; genomic RNA; improved; innovation; innovative technologies; insight; meter; next generation; novel; particle; prevent; protein distribution; recruit; superresolution imaging; targeted treatment; tomography; two-photon

Project Summary/Abstract Understanding the nature of the molecular interactions involved in human immunodeficiency virus type 1 (HIV- 1) replication continues to provide important insights into the fundamental nature of retrovirus replication. Beyond the importance of such basic science investigations in addressing crucial knowledge gaps in the field, such studies can inform antiretroviral target identification, and have broad applications towards therapy and cure. One of the key aspects of HIV-1 replication that has remained underexplored has been virus particle assembly. The reasons for this have included the challenges associated with the detailed behavior of Gag translocation to the plasma membrane, the engagement of particle assembly sites, the molecular interactions that drive virus particle assembly, and subsequent particle biogenesis and morphology. Integrative, comparative retrovirology has been particularly informative in gaining a deeper understanding of these steps in retroviral replication. For instance, immature particle morphology and the flexibility of the immature Gag lattice differs among retroviruses, which have been insightful for understanding the role of gaps in the Gag lattice and its relationship to relieving stress of lattice curvature. Furthermore, the recruitment pathways for retroviral Gag punctum formation, as well as the role and nature of the actin cortex on assembly site selection and biogenesis also remain poorly understood aspects of the retrovirus assembly pathway. In this application, we propose to investigate comparative analysis of HIV particle morphology and particle biogenesis through innovative state-of-the-art experimental approaches. In particular, we propose to employ cryo-electron microscopy/tomography (cryo-EM/ET), total internal reflection fluorescence (TIRF) microscopy, photoactivated localization microscopy (PALM), dual-color z-scan imaging and double helix-point spread function (DH-PSF) imaging to perform 3D super-resolution imaging and 3D single particle tracking in living cells to investigate 1) high-resolution comparative structural analysis HIV particle morphology and maturation, 2) investigate Gag puncta biogenesis at particle assembly sites, and 3) investigate the role of the actin cortex as a physical barrier in human retrovirus particle assembly. These novel studies harness innovative technologies in order to provide new insights into a highly significant yet poorly understood aspect of the HIV-1 life cycle, address important knowledge gaps in the field, provide insights into potential targets for therapeutic intervention, and inform efforts toward next-generation HIV therapies.

项目摘要/摘要 了解人类免疫缺陷病毒1型的分子相互作用的性质（HIV- 1）复制继续为逆转录病毒复制的基本本质提供重要的见解。超过 这种基础科学调查在解决该领域的关键知识差距方面的重要性 研究可以为抗逆转录病毒靶标识别提供信息，并在治疗和治疗方面有广泛的应用。一 HIV-1复制的关键方面一直保持不足，这是病毒颗粒组件。这 这样做的原因包括与堵塞易位的详细行为相关的挑战 质膜，颗粒组装位点的参与，驱动病毒颗粒的分子相互作用 组装以及随后的颗粒生物发生和形态。综合，比较逆转录病毒学一直是 在对逆转录病毒复制中的这些步骤更深入地了解这些步骤方面特别有用。例如， 未成熟的粒子形态和未成熟的堵嘴晶格的柔韧性在逆转录病毒之间有所不同，这 对理解差距在插科打晶格中的作用及其与压力的关系很有见解 晶格曲率。此外，逆转录病毒堵嘴形成的招聘途径以及 肌动蛋白皮质在组装位点选择和生物发生上的作用和性质也保持不佳 逆转录病毒组装途径的各个方面。在此应用中，我们建议研究比较分析 通过创新的最先进的实验方法，HIV颗粒形态和粒子生物发生。 特别是，我们建议采用冷冻电子显微镜/断层扫描（Cryo-EM/ET），总内部反射 荧光（TIRF）显微镜，光活化定位显微镜（棕榈），双色Z-SCAN成像和 双螺旋点扩展功能（DH-PSF）成像以执行3D超分辨率成像和3D单一 活细胞中的粒子跟踪研究1）高分辨率比较结构分析HIV粒子 形态学和成熟，2）研究颗粒组装位点的GAG PuncTA生物发生，3）研究 肌动蛋白皮质作为人逆转录病毒颗粒组件中物理屏障的作用。这些新型研究 利用创新技术，以便为一种备受理解的知名度提供新的见解 HIV-1生命周期的方面，解决该领域的重要知识差距，提供有关潜在潜力的见解 治疗干预的目标，并为下一代艾滋病毒疗法提供努力。