Structural Protein Networks ("Interactome") in Herpesviruses

描述(申请人提供)：疱疹病毒病毒粒子由四个结构元素组成：中央核心的一个大的双链DNA分子；包裹基因组的二十面体衣壳；紧邻衣壳的称为被膜的一层；以及包裹整个结构并嵌入病毒糖蛋白的外膜或包膜。感染性颗粒的组装和成熟是一个复杂的过程，也是一个特征不佳的过程。这个项目的目标是利用一种新开发的蛋白质芯片方法来阐明疱疹病毒中病毒粒子的组装过程。我们的工作假设是，病毒的组装和成熟是一个连续的过程，依赖于多蛋白复合体之间的有序相互作用。我们将重点分析来自甲型疱疹病毒(HSV-1)、β疱疹病毒(HCMV和HHV-7)和伽马疱疹病毒(KSHV和EBV)家族的23个保守结构蛋白之间的蛋白质相互作用，以及另外两个HSV-1特有的蛋白质，共计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，并应用一种创新的方法来识别多水平的蛋白质相互作用。此外，我们计划使用上述蛋白质芯片鉴定保守的疱疹病毒编码的蛋白激酶UL13、ORF36、BGLF4、UL97和U69的特定底物以及甲型疱疹病毒保守的蛋白激酶US3的底物，并确定所识别的底物的磷酸化将在多大程度上影响它们与其他蛋白质/蛋白质复合体的相互作用能力。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，这将提供对疱疹病毒中病毒粒子组装的序列和顺序的洞察。疱疹病毒是人类的主要病原体，会导致终生持续感染，并导致从轻微的冻疮到眼角膜炎甚至癌症的各种临床表现。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这些感染的有效治疗方法非常重要。上述研究将产生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.