HTLV-1 Particle Analysis and Gag Interactions

HTLV-1 颗粒分析和堵嘴相互作用

基本信息

批准号：
8371118
负责人：
Louis M Mansky
金额：
$ 33.14万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-28 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8371118
关键词：
Adult Binding Biogenesis Biological Models Caliber Cancer Etiology Caribbean region Cell Culture Techniques Cell membrane Cells Central Africa Cryoelectron Microscopy Cytoplasm Disease Experimental Models Fluorescence Fluorescence Microscopy Gagging General Population Goals HIV-1 Human Human T-Cell Leukemia Viruses Human T-lymphotropic virus 1 Individual Infection Inflammatory Interdisciplinary Study Japan Lead Life Membrane Molecular Nature Particle Size Prevalence Process Retroviridae Scanning Transmission Electron Microscopy Procedures Spectrum Analysis Spinal Cord Diseases Syndrome System T-Cell Leukemia Technology Testing Therapeutic Time Tropical Spastic Paraparesis Viral Virion Virus Virus Assembly Work base insight leukemia/lymphoma monomer novel novel therapeutics particle research study single molecule stoichiometry tissue culture tomography

项目摘要

DESCRIPTION (provided by applicant): Human T-cell leukemia virus (HTLV-1) infects about 20 million individuals worldwide and is the etiological agent of an adult T-cell leukemia/lymphoma (ATLL), and can also result in an inflammatory disease syndrome called HTLV-1-associated myelopathy (HAM)/tropical spastic paraparesis (TSP). Prevalence rates for HTLV-1 infection in the general population are greater than 1% in the Caribbean Basin, Central Africa, and South Japan. HTLV-1 is notorious for being difficult to study in cell culture, which ha 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. Using a tractable model system, state-of-the-art biophysical approaches, and an interdisciplinary research team, we have made novel observations that form the basis for this proposal. In this application, we propose to investigate questions related to HTLV-1 particle size, Gag stoichiometry in particles, and HTLV-1 Gag interactions in living cells using multiple experimental approaches. In particular, we will apply cryo-electron microscopy/tomography (cryo-EM/ET), total internal reflection fluorescence (TIRF) microscopy, and the novel single-molecule technology of fluorescence fluctuation spectroscopy (FFS) to investigate questions related to 1) particle size and Gag stoichiometry, 2) Gag targeting to membrane, and, 3) HTLV-1 particle biogenesis. The results from these proposed studies should provide further insight into fundamental aspects of HTLV-1 and retrovirus particle assembly, which may aid in developing therapeutics. PUBLIC HEALTH RELEVANCE: Human T-cell leukemia virus type 1 (HTLV-1) is a cancer-causing human retrovirus that infects about 20 million individuals worldwide, with prevalence rates greater than 1% in certain regions. 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. Such information may inform new therapeutic strategies.

描述（由申请人提供）：人类 T 细胞白血病病毒 (HTLV-1) 感染全球约 2000 万人，是成人 T 细胞白血病/淋巴瘤 (ATLL) 的病原体，也可导致炎症性疾病称为 HTLV-1 相关脊髓病 (HAM)/热带痉挛性截瘫 (TSP) 的综合征。在加勒比盆地、中非和日本南部，一般人群中 HTLV-1 感染的患病率大于 1%。 HTLV-1 因难以在细胞培养中进行研究而臭名昭著，这阻碍了对这些病毒如何在细胞中复制的严格分析，包括逆转录病毒组装所涉及的步骤。即使对于其他更容易处理的逆转录病毒系统，人们对逆转录病毒颗粒组装如何发生的细节也知之甚少。利用易于处理的模型系统、最先进的生物物理方法和跨学科研究团队，我们做出了新颖的观察，这些观察构成了该提案的基础。在此应用中，我们建议使用多种实验方法研究与 HTLV-1 粒径、颗粒中 Gag 化学计量以及活细胞中 HTLV-1 Gag 相互作用相关的问题。特别是，我们将应用冷冻电子显微镜/断层扫描（cryo-EM/ET）、全内反射荧光（TIRF）显微镜和荧光涨落光谱（FFS）的新型单分子技术来研究与1）相关的问题颗粒大小和 Gag 化学计量，2) Gag 靶向膜，以及 3) HTLV-1 颗粒生物发生。这些拟议研究的结果应该可以进一步深入了解 HTLV-1 和逆转录病毒颗粒组装的基本方面，这可能有助于开发治疗方法。公共卫生相关性：人类 T 细胞白血病病毒 1 型 (HTLV-1) 是一种致癌的人类逆转录病毒，感染全球约 2000 万人，在某些地区的患病率超过 1%。 HTLV-1 组装的基础研究将带来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。这些信息可能会为新的治疗策略提供信息。