Structural Protein Networks ("Interactome") in Herpesviruses

描述（由申请人提供）：疱疹病毒病毒粒子由四个结构元件组成：中央核心中的大双链DNA分子;二十面体衣壳，其包围基因组;直接包围衣壳的层，称为被膜;以及外膜或包膜，其包围整个结构，其中嵌入病毒糖蛋白。感染性颗粒的组装和成熟是一个复杂且表征不佳的过程。本计画的目的是利用蛋白质晶片的方法来探讨疱疹病毒的病毒粒子组装过程。我们的工作假设是，病毒组装和成熟是一个顺序的过程，是依赖于有序的多蛋白质复合物之间的相互作用。我们将重点分析来自α疱疹病毒（HSV-1），β疱疹病毒（HCMV和HHV-7）和γ疱疹病毒（KSHV和EBV）家族的23个保守结构蛋白之间的蛋白质相互作用，以及HSV-1特异性的两个额外蛋白质，共117个结构蛋白。我们将构建包含117种蛋白质的疱疹病毒蛋白质芯片，并应用创新的方法在多个水平上鉴定蛋白质相互作用。此外，我们计划使用上述蛋白芯片鉴定疱疹病毒编码的保守蛋白激酶（UL 13、ORF 36、BGLF 4、UL 97和U69）以及α疱疹病毒保守蛋白激酶US 3的特异性底物，并确定所鉴定底物的磷酸化在多大程度上影响它们与其他蛋白/蛋白复合物相互作用的能力。所有数据的整合将使我们能够建立一个复杂的蛋白质相互作用网络，这将为疱疹病毒中病毒体组装的序列和顺序提供见解。疱疹病毒是主要的人类病原体，其引起终身持续感染，并导致从轻度唇疱疹到眼角膜炎甚至癌症的临床表现。因此，由于这些病毒引起的感染是一个主要的公共卫生问题，了解疱疹病毒的生物学对于开发这些感染的有效治疗是重要的。上述研究的主要实际意义是鉴定可用于开发病毒特异性抗病毒药物的基本相互作用。

DESCRIPTION (provided by applicant): The herpesvirus virion is comprised of four structural elements: a large double-stranded DNA molecule in the central core; an icosahedral capsid, which encloses the genome; a layer that immediately surrounds the capsid termed the tegument; and an outer membrane or envelope, which encloses the whole structure and in which are embedded the viral glycoproteins. The assembly and maturation of the infectious particle is a complex and poorly characterized process. The goal of this project is to elucidate the process of virion assembly in herpesviruses using a newly developed protein chip approach. Our working hypothesis is that virus assembly and maturation is a sequential process and is dependent on ordered interactions between multi-protein complexes. We will focus on analyzing protein interactions among 23 conserved structural proteins from each of alphaherpesvirus (HSV-1), betaherpesvirus (HCMV and HHV-7), and gammaherpesvirus (KSHV and EBV) families as well as two additional proteins specific for HSV-1, a total of 117 structural proteins. We will construct herpesvirus protein chips containing the 117 proteins and apply an innovative approach for identifying protein interactions at multiple levels. Further, we plan to identify the specific substrates of the conserved herpesvirus-encoded protein kinase (UL13, ORF36, BGLF4, UL97, and U69) as well as those of the alphaherpesvirus conserved protein kinase US3 using the above protein chips, and determine to what extent the phosphorylation of the identified substrates will affect their abilities to interact with other proteins/protein complexes. The integration of all the data will allow us to build a complex protein interaction network that will provide insights into the sequence and order of the virion assembly in herpesviruses. Herpesviruses are major human pathogens that cause life-long persistent infections and result in clinical manifestations that range from a mild cold sore, to ocular keratitis and even cancer. Thus, infections due to these viruses are a major public health concern and understanding the biology of herpesviruses is important in the development of efficacious treatments of these infections. The major practical significance that will result from the above studies is the identification of essential interactions that can be used to develop virus- specific antivirals.