Proteins Mediating Interaction of HIV-1 and JCV in CNS

介导中枢神经系统中 HIV-1 和 JCV 相互作用的蛋白质

• 批准号: 6450231
• 负责人: EDWARD M. JOHNSON
• 金额: $ 37.17万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-02-01 至 2007-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6450231
• 关键词: AIDS; AIDS /HIV neuropathy; Polyomavirus hominis 2; central nervous system; cyclins; electron microscopy; gene expression; genetic transcription; genome; glia; human immunodeficiency virus 1; human tissue; latent virus infection; molecular genetics; neurotropic virus; oligodendroglia; point mutation; postmortem; progressive multifocal leukoencephalopathy; protein structure function; virus RNA; virus genetics; virus infection mechanism; virus protein; virus replication; virus virus interaction

DESCRIPTION: (provided by applicant) JC virus (JCV) is the etiologic agent of the neurodegenerative disease, progressive multifocal leukoencephalopathy (PML). JCV is normally latent in non-immunocompromised people, but it is activated in brains of individuals with AIDS. Because of the high incidence of PML in AIDS, we have hypothesized that activation of JCV in glial cells of the brain is influenced by HIV-1 infection. We have found that JCV late gene transcription is stimulated by the HIV-1 Tat protein through action at sequence elements bound by the cellular protein, Pur about. Tat and Pur0 about form a highly specific complex, and this complex acts to stimulate transcription at both the HW-1 TAR RNA element and the JCV late promoter. The complex also interacts with T-antigen to enhance JCV DNA replication. Purc about is a frequent partner of cyclin/CDK complexes, as is another Tat-binding protein, cyclin T1. This proposal aims to continue to exploit the expertise of two independent laboratories to coordinate studies on HIV-1, JCV and cellular proteins, including Pur0 about and cyclin T1. We shall take advantage of rapid autopsy procedures at the Manhattan HIV Brain Bank to perform immunogold electron microscopy on PML brain samples to colocalize Tat with these cellular proteins. We shall elucidate the dynamics of the interactions between Tat, T-antigen and Puro about during the course of JCV infection of oligodendrocytes and determine whether transcriptional activation involves activity of cyclin T1/CDK9. We shall characterize the involvement of Purc about or cyclin T1/CDK9 in Tat activation of transcription at TAR(+) or TAR(-) HIV-1 LTR promoters. Use of Puro about point mutants in AA V vectors will help dissect the contributions of different molecular unctional groups to HIV-1 replication in microglial/monocytic cells. We shall pursue our finding that exogenous Tat at low concentrations is incorporated by KG-1 oligodendrocytes and stimulates DNA replication initiated at the JCV origin. We shall determine the mechanism by which Tat enhances replication initiated at the JCV origin in human oligodendrocytes both in vivo and using a new in vitro system. Results will help elucidate pathways of activation of HIV-1 and JCV in the brain and will help target particular molecular interactions for therapy.

描述：（由申请人提供）JC 病毒（JCV）是下列疾病的病原体： 神经退行性疾病，进行性多灶性白质脑病 （PML）。 JCV 通常在非免疫功能低下的人群中呈潜伏状态，但 在艾滋病患者的大脑中被激活。由于发病率高 在 AIDS 中的 PML 中，我们假设，JCV 的激活是在 大脑受到 HIV-1 感染的影响。我们发现JCV晚期基因 HIV-1 Tat 蛋白通过序列作用刺激转录 与细胞蛋白质结合的元素，Pur 左右。 Tat 和 Pur0 大约形成一个 高度特异性的复合物，该复合物的作用是刺激转录 HW-1 TAR RNA 元件和 JCV 晚期启动子。该综合体还 与 T 抗原相互作用，增强 JCV DNA 复制。 Purc大约是一个 细胞周期蛋白/CDK 复合物的常见伙伴，也是另一种 Tat 结合蛋白， 细胞周期蛋白T1。该提案旨在继续利用两个人的专业知识 独立实验室协调 HIV-1、JCV 和细胞研究 蛋白质，包括Pur0和细胞周期蛋白T1。我们将利用快速 曼哈顿艾滋病脑库进行尸检程序以进行免疫金治疗 对 PML 大脑样本进行电子显微镜观察，以将 Tat 与这些细胞共定位 蛋白质。我们将阐明 Tat 之间相互作用的动态， JCV感染少突胶质细胞过程中T抗原和Puro的关系 并确定转录激活是否涉及细胞周期蛋白的活性 T1/CDK9。我们将描述 Purc 或细胞周期蛋白 T1/CDK9 的参与 TAR(+) 或 TAR(-) HIV-1 LTR 启动子处转录的 Tat 激活。使用 Puro 关于 A V 载体中点突变的研究将有助于剖析这些贡献 不同分子功能基团对 HIV-1 复制的影响 小胶质细胞/单核细胞。我们将追求我们的发现，即外源 Tat 在 低浓度被 KG-1 少突胶质细胞掺入并刺激 DNA 复制在 JCV 起点开始。我们将通过以下方式确定机制： Tat 增强了人类 JCV 起点起始的复制 少突胶质细胞在体内和使用新的体外系统。结果将 帮助阐明大脑中 HIV-1 和 JCV 的激活途径，并将 帮助针对特定的分子相互作用进行治疗。