HTLV Gag trafficking in living cells

描述（由申请人提供）：人类t细胞白血病病毒1型（HTLV-1）因难以在细胞培养中研究而臭名昭著，这阻碍了对这些病毒如何在细胞中复制的严格分析，包括涉及逆转录病毒组装的步骤。逆转录病毒颗粒组装发生的细节即使对于其他更容易处理的逆转录病毒系统也知之甚少。本提案寻求资金，以研究两个问题相关的Gag贩运活细胞使用一个令人兴奋的新的生物物理方法。我们将在活细胞中应用新颖的单分子荧光波动光谱技术（FFS）来研究1)wt Gag的HTLV-1 Gag寡聚化和病毒出芽结合蛋白缺陷的突变体，以及RNA包装缺陷的Gag突变体，以及2)Gag的迁移性和与膜的结合亲和力。这些拟议研究的结果将为HTLV-1和逆转录病毒颗粒组装的基本方面提供进一步的见解。公共卫生相关性：HTLV-1组装的基础研究将导致有关这些过程的详细信息，这将有助于更好地了解这些病毒如何在细胞中复制。

DESCRIPTION (provided by applicant): Human T-cell leukemia viruses type 1 (HTLV-1) is notorious for being difficult to study in cell culture, which has prohibited a rigorous analysis of how these viruses replicate in cells, including the steps involved in retrovirus assembly. The details for how retrovirus particle assembly occurs are poorly understood even for other more tractable retroviral systems. This proposal seeks funding to investigate two questions related to Gag trafficking in living cells using an exciting new biophysical approach. We will apply the novel single-molecule technology of fluorescence fluctuation spectroscopy (FFS) in living cells to investigate 1) HTLV-1 Gag oligomerization of wt Gag and mutants that have defects in binding proteins involved in virus budding as well as Gag mutants that have RNA packaging defects, and 2) Gag mobility and binding affinity to membrane. The results from these proposed studies should provide further insight into fundamental aspects of HTLV-1 and retrovirus particle assembly. PUBLIC HEALTH RELEVANCE: Fundamental studies of HTLV-1 assembly will lead to detailed information about these processes that will be useful for a better understanding of how these viruses replicate in cells.