HSC 70 AND AGNOPROTEIN IN PAPOVAVIRUS ASSEMBLY

乳腺病毒组装中的 HSC 70 和腺苷蛋白

• 批准号: 2057693
• 负责人: TIMOTHY P CRIPE
• 金额: $ 9.29万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-08-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2057693
• 关键词: 3T3 cells; Escherichia coli; Papovaviridae; Polyomavirus; binding proteins; capsid; gene deletion mutation; human papillomavirus; immunoprecipitation; intermolecular interaction; molecular chaperones; plasmids; protein folding; protein structure function; recombinant proteins; simian virus 40; stress proteins; transfection /expression vector; transport proteins; virus assembly; virus genetics; virus protein; viruslike particle

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.

基因治疗有望成为一种新的治疗方法， 疾病，但基因转移的最佳载体仍然难以捉摸。一种新型 目前受欢迎的基因递送策略的替代方案是体外 使用纯化的将核酸包装成病毒样颗粒 重组病毒衣壳，这可能会规避问题， 传统的基因转移这项建议旨在发展一个这样的模式 系统中使用了研究充分的乳多空病毒衣壳蛋白。纯化 重组VP1，鼠多瘤病毒和猴多瘤病毒的主要外壳蛋白 病毒40（SV40），和纯化的重组L1，主要外壳蛋白的 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，病毒基因组或外源DNA的包装在 使用VP1或L1的体外试验不成功。DNA双链化可能需要 病毒或细胞伴侣蛋白以稳定亚壳粒 组装过程中的中间体。因此，病毒外壳的相互作用 在病毒感染的细胞和体外， 将研究衣壳组装反应。最初的目的是 该项目是生物化学和遗传特性的体外 乳多空病毒外壳蛋白与细胞伴侣hsc 70的相互作用 和病毒伴侣未知蛋白。该项目还将确定在 hsc7O在衣壳蛋白核转运中的体内作用以及 未知蛋白和HSC 70对体外病毒组装的影响。最后如果 感染性病毒样颗粒在体外被成功地重建， 将评估它们的基因转移能力。除了 阐明病毒组装的机制，该项目可能会导致一个新的 在未来基因治疗中潜在有用基因递送系统 协议.