HTLV-1 Particle Analysis and Gag Interactions

HTLV-1 颗粒分析和堵嘴相互作用

• 批准号: 8537954
• 负责人: Louis M Mansky
• 金额: $ 27.28万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 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/8537954
• 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

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.

描述（由申请人提供）：人类T细胞白血病病毒（HTLV-1）在全球范围内感染约2000万个个体，是成年T细胞白血病/淋巴瘤（ATLL）的病因学药，也可能导致称为HTLV-1相关骨髓病（Ham）骨骼疾病（Ham）/Tropical parapical Paraparaical（Trop）（TRAPARPARAPARAPIAL）/TROPARPARAPILAPILAPILAL（TROPARPARPARAPILAPILAPILAPILAPILAL）（TROP）。在加勒比海盆地，中非和日本，普通人群中HTLV-1感染的患病率大于1％。 HTLV-1因在细胞培养中难以研究而臭名昭著，该细胞培养禁止对这些病毒在细胞中复制的严格分析，包括逆转录病毒组装中涉及的步骤。逆转录病毒颗粒组件的细节即使对于其他更可拖动的逆转录病毒系统也是很差的。使用可拖动的模型系统，最先进的生物物理方法和跨学科研究团队，我们进行了新的观察，构成了该建议的基础。在此应用中，我们建议使用多种实验方法研究与HTLV-1粒径，颗粒中的GAG化学计量和HTLV-1 GAG相互作用有关的问题。特别是，我们将采用冷冻电子显微镜/断层扫描（Cryo-EM/ET），总内反射荧光（TIRF）显微镜以及新型的荧光波动光谱谱（FFS）的单分子技术来研究与1个粒度和gag sToichiChiometry，2）粒子靶标的粒度和gag tosing tombrane and embrane，3），3）。这些拟议的研究的结果应提供对HTLV-1和逆转录病毒颗粒组装基本方面的进一步见解，这可能有助于开发治疗疗法。