Analysis of human retrovirus particle assembly sites

人逆转录病毒颗粒组装位点分析

• 批准号: 10662501
• 负责人: Nora Willkomm
• 金额: $ 5.6万
• 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 2025-09-26
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10662501
• 关键词: Acquired Immunodeficiency Syndrome; Actins; Address; Appearance; Biogenesis; Biological; Cell Communication; Cell membrane; Cells; Child; Communities; Consumption; Cytoplasm; Cytoskeleton; Development; Electron Microscopy; Event; Fluorescence; Fluorescence Microscopy; Generations; Genetic Materials; HIV-1; Human; Human Milk; Human T-Cell Leukemia Viruses; Human T-lymphotropic virus 1; Imaging Techniques; Individual; Infection; Investigation; Knowledge; 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; Productivity; 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; Visualization; 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; particle; permissiveness; polarized cell; postnatal; recruit; transmission process; viral pandemic; 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 粘膜表面是人类逆转录病毒的绝大多数传输事件，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 通过口腔的感染是从母亲到儿童的产后传播的重要门户。 病毒通过细胞到细胞传播传播有助于在新感染的个体中建立病毒感染。 尽管我们对逆转录病毒复制的理解取得了重大进步，但有很多 细节仍然很少理解。例如，目前尚不清楚病毒颗粒组装位点如何 确定以及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 细胞向细胞传输。在此应用中，我建议使用分子病毒学和最先进的成像 通过比较阐明不同人逆转录病毒的细胞到细胞传播机制的方法 分析。有充分的文献证明，HTLV-1通过细胞 - 细胞触点有效地传播，即病毒学 突触（VS）。 HIV-1可能也是如此，但通常被低估了。病毒传播 通过形成VS的细胞 - 细胞接触可以导致极化病毒颗粒释放到Vs中。病毒 颗粒形成是由插入结构蛋白驱动的，该蛋白质在病毒组装位点进行了多种形式（例如， 细胞接触点），导致颗粒生物发生和释放。在初步研究中，我们的实验室观察到 主要从等离子体募集了在非极化细胞中使用的GAG池 HTLV-1的膜，而用于HIV-1 GAG的膜是从细胞质中募集的。对 插入颗粒组装位点的插科打nirial模式可能有助于解释细胞至细胞的独特依赖 与HIV-1相比，细胞传播是HTLV-1的病毒扩散模式。我建议2行 该应用的投资。我将首先研究HIV-1和HTLV-1 GAG Puncta的差异是否存在 在非极化细胞中观察到的生物发生也在极化细胞中配置。其次，我将研究病毒 - 有助于促进人逆转录病毒组装的宿主细胞相互作用，尤其是在细胞 - 细胞接触的情况下， 这在口腔生物学中特别重要。这个目标的一个重要方面将是使用小说和 创新技术，低温相关的光和电子显微镜（Cryo-Clem），以获得更大的 对宿主细胞蛋白对病毒组装重要的作用的见解。人逆转录病毒颗粒组件 是感染性病毒传播的关键步骤，包括在粘膜表面的口服传播 细胞电池触点。这些研究将有助于贡献新信息，以更好地了解 人类逆转录病毒复制，这些复制未完全理解并代表病毒学领域的知识差距 并将提供关键信息，以帮助预防口腔逆转录病毒传播。