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: 6622545
• 负责人: EDWARD M. JOHNSON
• 金额: $ 36.5万
• 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/6622545
• 关键词: 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：(申请人提供)JC病毒(JCV)是 神经退行性疾病，进行性多灶性白质脑病 (PML)。JCV在非免疫功能低下的人中通常是潜伏的，但它是 在艾滋病患者的大脑中被激活。因为这种疾病的高发病率 PML在AIDS中，我们假设JCV在脑胶质细胞中的激活 大脑受到HIV-1感染的影响。我们已经发现了JCV晚期基因 HIV-1 Tat蛋白通过作用于序列来刺激转录 被细胞蛋白质结合的元素，无处不在。关于表格a的Tat和Pur0 高度特异的复合体，该复合体刺激转录 HW-1 TAR RNA元件和JCV晚期启动子。该建筑群还 与T抗原相互作用，增强JCV DNA复制。Purc About是一种 Cyclin/CDK复合体的频繁伙伴，以及另一种TAT结合蛋白， 细胞周期蛋白T1。这项提议旨在继续利用两家公司的专长 独立实验室协调对艾滋病毒-1、JCV和细胞的研究 蛋白质，包括Pur0约和细胞周期蛋白T1。我们将利用RAPID 曼哈顿艾滋病毒脑库的尸检程序进行免疫金 PML脑组织标本TAT与这些细胞共定位的电子显微镜观察 蛋白质。我们将阐明TAT和TAT之间相互作用的动力学。 少突胶质细胞感染JCV过程中T抗原与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与细胞周期蛋白T1/CDK9的关系 在TAT激活转录的TAR(+)或TAR(-)HIV-1LTR启动子。使用 Puro关于AA V载体中的点突变的研究将有助于剖析其贡献 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。我们将继续寻找外源性纹身 KG-1寡突胶质细胞掺入低浓度KG-1刺激DNA 复制在JCV源处启动。我们将通过以下方式确定该机制 哪种TAT可增强人类JCV起始点的复制 体内和使用一种新的体外系统的少突胶质细胞。结果将 帮助阐明HIV-1和JCV在大脑和意志中的激活途径 帮助针对特定的分子相互作用进行治疗。