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素组成：中央核心有一个大的双链DNA分子；二十面体衣壳，包裹着基因组；直接包围衣壳的一层，称为被皮；外膜或包膜，包裹着整个结构，其中嵌入了病毒糖蛋白。感染颗粒的组装和成熟是一个复杂且特征不明确的过程。本项目的目的是利用一种新开发的蛋白质芯片方法阐明疱疹病毒的病毒粒子组装过程。我们的工作假设是病毒的组装和成熟是一个连续的过程，依赖于多蛋白复合物之间有序的相互作用。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个保守结构蛋白以及另外两个HSV-1特异性蛋白之间的蛋白相互作用，共117个结构蛋白。我们将构建包含117种蛋白的疱疹病毒蛋白芯片，并应用一种创新的方法来识别蛋白在多个水平上的相互作用。此外，我们计划使用上述蛋白芯片鉴定保守疱疹病毒编码蛋白激酶（UL13, ORF36, BGLF4， UL97和U69）的特定底物以及α疱疹病毒保守蛋白激酶US3的底物，并确定鉴定的底物磷酸化将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，这将为疱疹病毒病毒粒子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，可引起终身持续性感染，并导致从轻度唇疱疹到眼部角膜炎甚至癌症的临床表现。因此，由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有效治疗这些感染的方法非常重要。上述研究的主要实际意义是确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.