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: 6701819
• 负责人: EDWARD M. JOHNSON
• 金额: $ 37.59万
• 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/6701819
• 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：（由申请人提供）JC病毒（JCV）是 神经退行性疾病，进行性多灶性白血病病 （PML）。 JCV通常在非免疫力低下的人中潜在 在有艾滋病的个体的大脑中激活。由于很高的发生率 PML在艾滋病中，我们假设JCV在胶质细胞中的激活 大脑受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 由细胞蛋白绑定的元素，周围。 tat和pur0关于形式 高度特定的复合 HW-1 TAR RNA元素和JCV晚期启动子均两者均。这是一个综合体 与T-抗原相互作用以增强JCV DNA复制。 purc是一个 Cyclin/CDK复合物的常见伴侣，另一种结合蛋白也是如此 细胞周期蛋白T1。该建议旨在继续利用两个的专业知识 独立实验室，以协调HIV-1，JCV和细胞的研究 蛋白质，包括有关PUR0和细胞周期蛋白T1的蛋白质。我们将利用快速 曼哈顿艾滋病毒脑库的尸检程序以进行免疫光 PML脑样品上的电子显微镜与这些细胞共定位 蛋白质。我们将阐明TAT之间相互作用的动态 在JCV感染过程中，T-抗原和Puro大约是少突胶质细胞 并确定转录激活是否涉及细胞周期蛋白的活性 T1/CDK9。我们将表征有关或细胞周期蛋白T1/cdk9的参与 在TAT（+）或TAR（ - ）HIV-1 LTR启动子处转录的TAT激活中。使用 puro关于AA V量中点突变体的围绕点突变体将有助于剖析贡献 在不同的分子非观点组中复制HIV-1 小胶质细胞/单核细胞。我们将追求我们发现的外源性TAT 低浓度由kg-1少突胶质细胞纳入并刺激DNA 在JCV Origin发起的复制。我们将通过 TAT增强了在人类JCV起源的复制 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激活的途径，将 帮助靶向特定的分子相互作用进行治疗。