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.

项目摘要 粘膜表面占人类逆转录病毒传播事件的绝大多数--例如人类 免疫缺陷病毒1型(HIV-1)和人类T细胞白血病病毒1型(HTLV-1)。HIV-1和HTLV-1 通过口腔的感染是产后母婴传播的重要途径。 通过细胞间传播的病毒有助于在新感染者中建立病毒感染。 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 仍然知之甚少的细节。例如，目前还不清楚病毒颗粒组装部位的情况。 确定了病毒结构蛋白和遗传物质如何转移到内部的这些位置 质膜的小叶。病毒颗粒组装的细节在以下情况下尤其未知 细胞到细胞的传输。在这个应用中，我建议使用分子病毒学和最先进的成像技术 用比较方法阐明不同人类逆转录病毒细胞间传播机制的方法 分析。众所周知，HTLV-1是通过细胞间接触，即病毒学上的 突触(VS)。艾滋病毒-1也可能是这种情况，但通常被低估了。病毒传播 通过形成VS的细胞-细胞接触可导致极化的病毒颗粒释放到VS中。病毒 颗粒的形成由在病毒组装部位多聚体的Gag结构蛋白驱动(例如， 细胞接触点)，导致颗粒的生物生成和释放。在初步研究中，我们的实验室观察到 非极化细胞中用于颗粒生物发生的GAG池主要从血浆中招募 HTLV-1的Gag是从细胞质中招募的，而HIV-1的Gag是从细胞质招募的。这一基本观察结果 GAG招募到颗粒组装部位的不同模式可能有助于解释细胞对-GAG的不同依赖 细胞传播是HTLV-1与HIV-1病毒传播的一种生产方式。我建议用两行字 这个应用程序的调查。我将首先调查HIV-1和HTLV-1之间的差异 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。第二，我会调查病毒- 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-细胞接触的背景下， 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将是使用小说和 创新技术，低温相关光学和电子显微镜(Cryo-Clem)，以获得更大的 洞察宿主细胞蛋白对病毒组装的重要作用(S)。人类逆转录病毒颗粒组装 是传染性病毒传播的关键步骤，包括在黏膜表面的口腔传播 细胞间的接触。这些研究将有助于提供新的信息，以更好地了解 尚未完全了解的人类逆转录病毒复制，代表着病毒学领域的知识空白 并将提供关键信息，帮助预防通过口腔的逆转录病毒传播。