HSC 70 AND AGNOPROTEIN IN PAPOVAVIRUS ASSEMBLY

基因疗法有望成为一种新的治疗方法 疾病，尽管基因转移的最佳载体仍然难以捉摸。一本小说 目前受欢迎的基因传递策略的替代方案是体外试验 用纯化的核酸包装成病毒样颗粒 重组病毒衣壳，这可能会绕过 传统的基因转移。这项提议旨在开发这样一种模式 使用研究得很好的乳头病毒衣壳蛋白的系统。纯净的 重组VP1，小鼠多瘤病毒和猿猴的主要外壳蛋白 病毒40(SV40)，以及纯化的重组L1，其主要外壳蛋白 人乳头瘤病毒(HPV)在体外自组装成病毒样 粒子。然而，到目前为止，将病毒基因组或外源DNA包装在 使用VP1或L1进行体外培养并不成功。DNA封装可能需要 稳定亚囊膜的病毒或细胞伴侣蛋白 装配过程中的中间体。因此，病毒外壳的相互作用 病毒感染细胞和体外含伴侣蛋白的蛋白质 将研究衣壳组装反应。这样做的最初目的是 项目是在体外进行生物化学和遗传学表征 乳头状病毒外壳蛋白与细胞伴侣hsc70的相互作用 和病毒伴侣不可知蛋白。该项目还将决定在 Hsc7O在衣壳蛋白核转运中的活体作用及对HSC7O的影响 不可知蛋白和hsc70对病毒的体外组装。最后，如果 在体外成功地重建了具有感染性的病毒样颗粒， 他们的基因转移能力将得到评估。除了……之外 阐明病毒组装的机制，这一项目可能导致一种新的 一种在未来基因治疗中可能有用的基因传递系统 协议。

Gene therapy holds promise as a new treatment approach for a variety of diseases, though optimal vehicles for gene transfer remain elusive. A novel alternative to currently favored gene delivery strategies is in vitro packaging of nucleic acid into virus-like particles using purified recombinant viral capsids, which might circumvent problems with conventional gene transfer. This proposal seeks to develop a such a model system using the well studied papovavirus capsid proteins. Purified recombinant VP1, the major coat protein of murine polyomavirus and simian virus 40 (SV40), and purified recombinant L1, the major coat protein of human papillomaviruses (HPV), self-assemble in vitro into virus-like particles. As yet, however, packaging of viral genomes or foreign DNA in vitro using VP1 or L1 has been unsuccessful. DNA encapsidation may require viral or cellular chaperone proteins to stabilize sub-capsomer intermediates during assembly. Therefore, the interaction of viral coat proteins with chaperone proteins in both virus-infected cells and in vitro capsid assembly reactions will be investigated. The initial aim of this project is to biochemically and genetically characterize the in vitro interactions of papovavirus coat proteins with the cellular chaperone hsc70 and viral chaperone agnoprotein. The project will also determine the in vivo role of hsc7O in capsid protein nuclear transport and the effect of the agnoprotein and hsc70 on in vitro viral assembly. Finally, if infectious virus-like particles are successfully reconstructed in vitro, their capacity for gene transfer will be assessed. In addition to elucidating mechanisms of viral assembly, this project may lead to a new system of gene delivery potentially useful in future gene therapy protocols.