Single Molecule HIV-1 NC/Gad-DNA Interactions

描述（由申请人提供）：本研究的目的是研究HIV-1和其他逆转录病毒Gag和核衣壳蛋白（NC）在调控逆转录病毒系统中核酸结构转变和病毒自组装过程中的作用。单分子DNA拉伸实验可以诱导DNA螺旋线圈转变。对逆转录病毒NC蛋白在这一转变中的作用的研究揭示了核酸伴侣活性的机制。我们将通过检测来自HIV-1和其他逆转录病毒的NC以及HIV-1 Gag蛋白对DNA包装和DNA螺旋螺旋转变的影响来研究这些相互作用。我们将使用这种新技术来探测药物对NC蛋白改变DNA融化能力的影响，也表明DNA和RNA的特定结构可以用力谱来探测。我们将扩展这项技术来研究逆转录病毒NC和Gag蛋白与特定RNA结构的相互作用，这些RNA结构对逆转录病毒复制很重要。然后，我们将开发逆转录病毒NC和Gag蛋白与核酸相互作用的定量模型，这将增加我们对核酸伴侣活性和病毒组装的理解。三个具体目标是：1。探讨逆转录病毒核衣壳蛋白的核酸伴侣活性和Gag蛋白的病毒自组装。2. 探讨NC结合药物改变NC核酸伴侣活性的能力。3. 目的探讨逆转录病毒核衣壳和Gag蛋白与HIV-1 RNA结构的相互作用。

DESCRIPTION (provided by applicant): The objective of this research is to investigate the role of HIV-1 and other retroviral Gag and nucleocapsid proteins (NC) in the regulation of processes involving nucleic acid structural transitions and viral self assembly in retroviral systems. Single molecule DNA stretching experiments can induce the DNA helix-coil transition. Investigation of the effects of retroviral NC proteins on this transition has shed light on the mechanism of nucleic acid chaperone activity. We will investigate these interactions by examining the effects of NC from HIV-1 and other retroviruses as well as HIV-1 Gag proteins on DNA packaging and the DNA helix-coil transition. We will use this novel technique to probe the effects of drugs on the capability of NC proteins to alter DNA melting, it has also been shown that specific structures of DNA and RNA can be probed using force spectroscopy. We will extend this technique to study the interactions of retroviral NC and Gag proteins with specific RNA structures that are important for retroviral replication. We will then develop a quantitative model for the interaction of retroviral NC and Gag proteins with nucleic acids, which will increase our understanding of nucleic acid chaperone activity and viral assembly. The 3 specific aims are: 1. To probe the nucleic acid chaperone activity of retroviral nucleocapsid proteins and the viral self-assembly of Gag proteins. 2. To probe the capabilities of NC binding drugs to alter NC's nucleic acid chaperone activity. 3. To probe the interaction of retroviral nucleocapsid and Gag proteins with HIV-1 RNA structures.