Assembly and Dissassembly of Animal Viruses

PI最近发现，可以使用从自然产生的空衣壳(即Proapsids)获得的14S五聚体亚基，在体外组装类似病毒的壳。该系统为设计能够形成脊髓灰质炎病毒的体外反应混合物提供了依据。这项建议的主要目的是建立这样的反应混合物，并将其用作纯化和分离存在于Hela细胞中的病毒特异性“形态生成”因子(S)的实验，该因子负责从体内14S物质中形成Proapsids(和病毒颗粒)。将进行以下实验：1)鉴定其性质；2)确定其作用模式；3)定量测定支持体外病毒组装系统所需的数量。另外两个系列的实验也在计划之中。第一个目的是表征病毒粒子和针对不同病毒衣壳表位的单抗之间相互作用的性质。第二部分涉及脊髓灰质炎病毒RNA中“内部”起始点的重要性。作为一个整体，该项目将大大有助于我们理解脊髓灰质炎病毒和其他小核糖核酸病毒的蛋白质的合成和加工方式。

The PI has recently found that virus-like shells can be assembled in vitro, using 14S pentameric subunits obtained from naturally-occuring empty capsids (i.e. procapsids). This system provides a basis for designing an in vitro reaction mixture capable of forming poliovirions. The main objective of this proposal is to establish such a reaction mixture and to use it as an assay to purify and isolate the viral-specific "morphopoietic" factor(s) present in Hela cells which is responsible for forming procapsids (and virus particles) from 14S material in vivo. Experiments will be conducted to: 1) identify its nature 2) ascertain its mode of action, and 3) quantitate the amount needed to support the in vitro virus assembly system. Two series of other experiments are also planned. The first is aimed at characterizing the nature of the interactions between the virions and monoclonal antibodies to different viral capsid epitopes. The second deals with the significance of the "internal" initiation site in polioviral RNA. The project as a whole will contribute substantially to our understanding of the way proteins of poliovirus and other picornaviruses are synthesized and processed.