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：
9333502
负责人：
Louis M Mansky
金额：
$ 47.16万
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 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9333502
关键词：
Adult Adult T-Cell Leukemia/Lymphoma Affect Age Antibodies Biogenesis Cancer Etiology Capsid Cell Culture Techniques Cell membrane Cell surface Cells Cryoelectron Microscopy Disease Etiology Event Female Fluorescence Fluorescence Microscopy Foundations Geographic Locations Goals HIV-1 Human Human Characteristics Human T-Cell Leukemia Viruses Human T-lymphotropic virus 1 Individual Inflammatory Investigation Knowledge Light Mason-Pfizer monkey virus Membrane Methods Microbial Biofilms Microscopy Molecular Molecular Models Morphology Nature Pathway interactions Population Prevalence Process Reporting Resolution Retroviridae Role Rous sarcoma virus Scanning Site Site-Directed Mutagenesis Spectrum Analysis Spinal Cord Diseases Structural Protein Structure Syndrome System Testing Translations Tropical Spastic Paraparesis Viral Virion Virus Virus Assembly Virus Replication aged comparative density experimental study innovation innovative technologies insight molecular modeling new technology novel novel therapeutic intervention particle reconstruction tomography transmission process viral transmission virological synapse virology virus envelope

项目摘要

Abstract Human T-cell leukemia virus (HTLV-1) infects about 15-20 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). HTLV-1 antibody prevalence rates vary among geographical areas, ranging from 0.2 to 10% among adults. This antibody prevalence increases with age, and can affect as much as 20 to 50% of the female population aged 60 and above. 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 like that of human immunodeficiency virus type 1 (HIV-1). For instance, recent evidence indicates that Gag-Gag interactions differ among retroviruses, which helps explain morphological differences that we have documented among immature retrovirus particles. Furthermore, the role for membrane-bound, non-punctate (np) Gag in the biogenesis of Gag puncta, as well as the nature of Gag puncta biogenesis in the context of cell-to-cell contacts also remain poorly understood aspects of the retrovirus assembly pathway. This is particularly for HTLV-1, for which we have found to have fundamentally distinct differences to that of HIV-1 regarding the role of membrane-bound np Gag in Gag punta biogenesis. In this application, we propose to continue our investigations on HTLV-1 immature and mature particle structure and particle biogenesis through innovative state-of-the-art experimental approaches with appropriate comparative analyses with HIV-1. In particular, we will apply cryo-electron microscopy/tomography (cryo-EM/ET), photoactivated localization microscopy (PALM), total internal reflection fluorescence (TIRF) microscopy and the novel technology of z-scan fluorescence fluctuation spectroscopy (FFS) in living cells to investigate 1) analysis of immature HTLV-1 Gag lattice structure, 2) the importance of membrane-bound, np Gag in HTLV-1 particle biogenesis, and 3) HTLV-1 particle biogenesis in the context of cell-cell contacts. Careful comparisons will be done with HIV-1. These novel studies harness innovative technologies in order to provide new insights into a highly significant and poorly understood aspect of the HTLV-1 particle assembly process, which is fundamentally distinct from that of HIV-1 and other retroviruses. Furthermore, our studies represent some of the most detailed studies conducted on the assembly of virus structural proteins, which provides fundamentally important information in virology on the molecular nature of virus particle assembly at the plasma membrane for enveloped viruses.

抽象的人类 T 细胞白血病病毒 (HTLV-1) 感染全球约 15-2000 万人，其病因是成人 T 细胞白血病/淋巴瘤 (ATLL) 的病原体，也可导致炎症性疾病综合征称为 HTLV-1 相关脊髓病 (HAM)/热带痉挛性截瘫 (TSP) 抗体流行率。不同地理区域的患病率有所不同，成人中这种抗体的患病率从 0.2% 到 10% 不等。随着年龄的增长，HTLV-1 可能会影响 20% 至 50% 的 60 岁及以上女性人群。因难以在细胞培养中研究而臭名昭著，这阻碍了对这些细胞如何进行严格分析病毒在细胞中复制，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，例如例如，最近的证据表明，1 型人类免疫缺陷病毒 (HIV-1) 是 Gag-Gag。逆转录病毒之间的相互作用不同，这有助于解释我们记录的形态差异此外，膜结合的非点状 (np) Gag 在未成熟逆转录病毒颗粒中的作用。 Gag puncta 的生物发生，以及细胞间接触背景下 Gag puncta 生物发生的性质还保留了逆转录病毒组装途径的不良方面，这对于 HTLV-1 来说尤其如此。我们发现它与 HIV-1 的作用有根本上的不同 Gag punta 生物发生中的膜结合 np Gag 在此应用中，我们建议继续我们的研究。通过创新研究HTLV-1未成熟和成熟颗粒结构和颗粒生物发生最先进的实验方法与 HIV-1 进行适当的比较分析。将应用冷冻电子显微镜/断层扫描 (cryo-EM/ET)、光激活定位显微镜 (PALM)、全内反射荧光 (TIRF) 显微镜和 z 扫描荧光新技术活细胞中的涨落光谱 (FFS) 研究 1) 未成熟 HTLV-1 Gag 晶格的分析结构，2) 膜结合的 np Gag 在 HTLV-1 颗粒生物发生中的重要性，以及 3) HTLV-1 将与 HIV-1 进行仔细比较。新颖的研究利用创新技术，为非常重要且重要的问题提供新的见解。 HTLV-1 粒子组装过程的一个方面人们知之甚少，该过程与 HTLV-1 粒子组装过程有根本不同 HIV-1 和其他逆转录病毒。此外，我们的研究代表了一些最详细的研究关于病毒结构蛋白的组装，这提供了病毒学的基本重要信息。包膜病毒质膜上病毒颗粒组装的分子性质。