Protein Interactions Involved in Orthopoxvirus Envelopment

描述(申请人提供)：痘病毒包括一大类DNA病毒，能够感染人类并导致疾病。虽然最臭名昭著的天花成员天花(天花的病原体)已从自然感染中根除，但仍有人担心在生物攻击期间秘密释放。此外，猴痘和其他家庭成员也提出了对可能导致流行病的流行性感染的担忧。痘病毒产生两种感染形式，细胞内成熟病毒(IMV)和细胞外病毒(EV)。IMV构成了子代病毒粒子的大部分。EV是由IMV的胞内包膜形成的，是细胞间传播和全身感染所必需的。目前只有8种病毒蛋白是EV形式所特有的。该项目的长期目标是了解正痘病毒包膜、运输和释放传染性EV的分子机制。此应用程序的直接目标是了解与三种主要EV糖蛋白以及它们之间的相互作用如何促进EV囊膜中适当的蛋白质含量，以及这如何影响全身感染。我们的具体目标是：1)精确定位EV蛋白质相互作用所需的特定残基，2)确定EV蛋白质相互作用之间的层次结构和空间关系。3)探讨EV蛋白相互作用与EV囊膜蛋白组成、感染性EV产生及体内致病机制的关系。这项研究的结论将提供关于三种主要的正痘病毒糖蛋白(A33、A34和B5)在分子、细胞和生物水平上相互作用的信息。所获得的结果将为深入了解痘病毒产生感染性EV并在其宿主内细胞间传播的分子机制提供更多的信息。此外，当构建疫苗和溶瘤平台的重组痘病毒载体时，它们将为智能设计决策提供信息。

DESCRIPTION (provided by applicant): Poxviruses include a large family of DNA viruses capable of infecting and causing disease in humans. While the most notorious member variola, the causative agent of smallpox, was eradicated from natural infection, there are still concerns about a clandestine release during a biological attack. In addition, monkeypox and other members of the family have raised concern about epizootic infections that are capable of causing epidemic. Poxviruses produce two infectious forms, intracellular mature virus (IMV) and extracellular virus (EV). IMV make up the majority of progeny virions. EV are formed by the intracellular envelopment of IMV and are required for cell-to-cell spread and systemic infection. Only 8 viral proteins are known to be unique to the EV form. The long-term goal of this project is to understand the molecular mechanisms employed by orthopoxoviruses to envelope, transport, and release infectious EV. The immediate goal of this application is to understand how interactions with, and amongst the three major EV glycoproteins facilitate proper protein content in the EV envelop and how this effects systemic infection. Our specific aims are; 1) Fine mapping of specific residues required for EV protein interaction, 2) To determine the hierarchy and spatial relationship between the EV protein interactions. 3) To determine the relationship between EV protein interaction EV envelope protein composition, infectious EV production, and pathogenesis in vivo. The conclusion of this study will provide information about the interaction between the three major orthopoxvirus glycoproteins (A33, A34, and B5) at the molecular, cellular and organismal levels. The results obtained will provide greater insight into the molecular mechanism poxviruses use to produce infectious EV and spread cell-to-cell in its host. In addition they will inform intelligent design decision when constructing recombinant poxvirus vectors for both vaccines and oncolytic platforms.