Proteins Mediating Interaction of HIV-1 and JCV in CNS

描述：（由申请人提供）JC病毒（JCV）是 进行性多灶性白质脑病 （PML）。JCV通常潜伏在非免疫功能低下的人群中，但它是 在艾滋病患者的大脑中被激活。由于发病率高， 在艾滋病的PML中，我们假设JCV在胶质细胞中的激活， 大脑受到HIV-1感染的影响。我们发现JCV晚期基因 HIV-1达特蛋白通过作用于 元素结合的细胞蛋白质，Pur约。达特和Pur 0约为a型 高度特异性的复合物，这种复合物的作用是刺激转录， HW-1 TAR RNA元件和JCV晚期启动子两者。该建筑群还 与T抗原相互作用以增强JCV DNA复制。采购大约是一个 细胞周期蛋白/CDK复合物的常见伴侣，如另一种Tat结合蛋白， 细胞周期蛋白T1。这项建议旨在继续利用两个专业知识， 独立实验室协调HIV-1、JCV和细胞 蛋白质，包括Pur 0和细胞周期蛋白T1。我们将利用快速 在曼哈顿艾滋病毒脑库进行免疫金的尸检程序 电子显微镜检查PML脑样品，以共定位达特与这些细胞 proteins.我们将阐明达特， JCV感染少突胶质细胞过程中T抗原和Puro的变化 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/CDK 9的参与， 在TAR（+）或TAR（-）HIV-1 LTR启动子处转录的达特激活中。使用 Puro关于AA V载体中的点突变体的研究将有助于剖析 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。我们将继续我们的发现，外源性达特， 低浓度被KG-1少突胶质细胞掺入并刺激DNA 复制起始于JCV起点。我们将通过以下方式确定机制： 该达特增强了人JCV起始处的复制 体内和使用新的体外系统的少突胶质细胞。结果将 有助于阐明HIV-1和JCV在大脑中的激活途径， 帮助靶向治疗特定的分子相互作用。

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.