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.

项目总结