Analysis of human retrovirus particle assembly sites

人逆转录病毒颗粒组装位点分析

• 批准号: 10326739
• 负责人: Nora Willkomm
• 金额: $ 4.52万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-27 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10326739
• 关键词: Acquired Immunodeficiency Syndrome; Actins; Address; Appearance; Biogenesis; Biological; Cell Communication; Cell membrane; Cells; Child; Communities; Consumption; Cytoplasm; Development; Electron Microscopy; Event; Fluorescence; Fluorescence Microscopy; Generations; Genetic Materials; HIV-1; Human; Human Milk; Human T-lymphotropic virus 1; Imaging Techniques; Individual; Infection; Investigation; Knowledge; Life; Location; Modification; Molecular; Molecular Virology; Mothers; Mucous Membrane; Nature; Neurons; Oral; Oral cavity; Oral mucous membrane structure; Pathway interactions; Patients; Pharmaceutical Preparations; Play; Prevention; Process; Protein Kinase C; Proteins; Recording of previous events; Reporting; Research; Research Personnel; Retroviridae; Role; Site; Source; Structural Protein; Surface; Testing; Ubiquitin; Vertical Disease Transmission; Viral; Viral Proteins; Viral Structural Proteins; Virion; Virus; Virus Assembly; Virus Diseases; casein kinase; comparative; cryogenics; effective therapy; experimental study; fluorescence imaging; gag Gene Products; imaging approach; innovative technologies; insight; light microscopy; new technology; novel therapeutic intervention; novel therapeutics; oral biology; pandemic disease; particle; polarized cell; postnatal; recruit; transmission process; viral transmission; virological synapse; virology

Project Summary Mucosal surfaces account for the vast majority of transmission events for human retroviruses – e.g., human immunodeficiency virus type 1 (HIV-1) and human T-cell leukemia virus type 1 (HTLV-1). HIV-1 and HTLV-1 infection through the oral cavity represents a significant gateway for postnatal transmission from mother to child. Virus spread via cell-to-cell transmission aids in establishment of viral infection in a newly infected individual. While significant advancements in our understanding of retrovirus replication have been made, there are many details that remain poorly understood. For instance, it is still unclear how virus particle assembly sites are determined and how viral structural proteins and genetic material translocate to these locations on the inner leaflet of the plasma membrane. The details of viral particle assembly are particularly unknown in the context of cell-to-cell transmission. In this application, I propose to use molecular virology and state-of-the-art imaging approaches to elucidate mechanisms of cell-to-cell transmission of different human retroviruses by comparative analyses. It is well documented that HTLV-1 is efficiently transmitted via cell-cell contacts, i.e., the virological synapse (VS). This is likely also the case for HIV-1 but has been commonly underappreciated. Virus transmission at cell-cell contacts via the formation of a VS can result in polarized virus particle release into the VS. Virus particle formation is driven by the Gag structural protein, which multimerizes at the virus assembly site (e.g., points of cell contact), resulting in particle biogenesis and release. In preliminary studies, our lab has observed that the pool of Gag utilized in particle biogenesis in non-polarized cells was primarily recruited from the plasma membrane for HTLV-1 whereas for HIV-1 Gag is recruited from the cytoplasm. This fundamental observation of differential modes of Gag recruitment to particle assembly sites may help explain the distinct reliance of cell-to- cell transmission as a productive mode of virus spread for HTLV-1 compared with that of HIV-1. I propose 2 lines of investigation for this application. I will first investigate whether the differences in HIV-1 and HTLV-1 Gag puncta biogenesis observed in non-polarized cells are also conserved in polarized cells. Second, I will investigate virus- host cell interactions that help facilitate human retrovirus assembly, particularly in the context of cell-cell contacts, which is of particular significance in oral biology. An important aspect of this aim will be the use of novel and innovative technology, cryogenic-correlative light and electron microscopy (cryo-CLEM), in order to gain greater insights into the role(s) of host cellular proteins important for virus assembly. Human retrovirus particle assembly is a critical step in infectious virus transmission, including oral transmission at mucosal surfaces in the context of cell-cell contacts. These studies will help contribute new information to better understand key aspects of human retroviral replication that are not fully understood and represent knowledge gaps in the field of virology and will provide critical information to aid in the prevention of retroviral transmission through the oral cavity.

项目概要 粘膜表面是人类逆转录病毒传播事件的绝大多数——例如，人类 1 型免疫缺陷病毒 (HIV-1) 和 1 型人类 T 细胞白血病病毒 (HTLV-1 和 HTLV-1)。 通过口腔感染是产后母婴传播的重要途径。 病毒通过细胞间传播传播有助于在新感染的个体中建立病毒感染。 虽然我们对逆转录病毒复制的理解已经取得了重大进展，但仍有许多问题 一些细节仍知之甚少，例如，目前尚不清楚病毒颗粒的组装位点是如何的。 确定了病毒结构蛋白和遗传物质如何易位到内部的这些位置 质膜的小叶，病毒颗粒组装的细节在背景下尤其未知。 在此应用中，我建议使用分子病毒学和最先进的成像技术。 通过比较阐明不同人类逆转录病毒的细胞间传播机制的方法 有充分证据表明 HTLV-1 通过细胞间接触（即病毒学）有效传播。 HIV-1 的情况也可能如此，但病毒传播通常被低估。 通过形成 VS 的细胞与细胞接触可导致极化病毒颗粒释放到 VS 中。 颗粒形成由 Gag 结构蛋白驱动，该蛋白在病毒组装位点发生多聚化（例如， 在初步研究中，我们的实验室观察到了颗粒的生物发生和释放。 非极化细胞中粒子生物发生中使用的 Gag 库主要是从血浆中招募的 HTLV-1 的 Gag 是从细胞质中招募的，而 HIV-1 的 Gag 是从细胞质中招募的。 Gag 招募到颗粒组装位点的不同模式可能有助于解释细胞与细胞之间的不同依赖性 与 HIV-1 相比，细胞传播是 HTLV-1 病毒传播的有效模式。 我将首先调查 HIV-1 和 HTLV-1 Gag puncta 是否存在差异。 在非极化细胞中观察到的生物发生在极化细胞中也是保守的。 其次，我将研究病毒。 宿主细胞相互作用有助于促进人类逆转录病毒组装，特别是在细胞与细胞接触的情况下， 这在口腔生物学中具有特别重要的意义，这一目标的一个重要方面是使用新颖的和 创新技术，低温相关光学和电子显微镜（cryo-CLEM），以获得更大的 深入了解宿主细胞蛋白对病毒组装的重要作用。 是传染性病毒传播的关键步骤，包括在粘膜表面的口腔传播 这些研究将有助于提供新信息以更好地理解细胞与细胞接触的关键方面。 人类逆转录病毒复制尚未完全理解，代表了病毒学领域的知识差距 并将提供重要信息以帮助预防逆转录病毒通过口腔传播。