Elucidation of genetic networks of HSV-1 virion proteins and discovery of their functions in the morphogenesis of the infectious virus particle

疱疹病毒是人类的病原体，会导致终身持续感染，并导致从轻微冻疮到癌症的各种临床表现。单纯疱疹病毒(HSV)是普通人群中最常见的病毒之一。感染HSV-1通常会导致口腔疾病，但也会导致眼部疾病(角膜炎)，导致失明或脑炎，这可能是致命的。2型感染的特征是生殖器溃烂疾病和新生儿感染。因此，单纯疱疹病毒引起的感染是一个主要的公共卫生问题，了解这种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。单纯疱疹病毒1型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被膜。就蛋白质组成和该结构成分所编码的功能而言，被膜是病毒粒子最复杂和最多样化的结构之一。在这一大群不同的病毒粒子中，并不是所有的蛋白质都被归类为实验室细胞培养系统中复制所必需的。所有的“非必需”蛋白质对病毒粒子成熟和排出有什么作用？这些“附属”基因产物可能在受感染的宿主中发挥重要作用，但对其功能的深入发现尚未全面完成。使用基因组装配线与合成基因组学方法相结合，我们可以通过在突变病毒中创造“合成致命性”来阐明这些蛋白质的功能。我们的工作假设是，通过对合成致死物的表型研究，我们可以开始建立这些“多余的”病毒粒子蛋白参与的途径，以及它们在HSV-1病毒粒子形态形成过程中的空间/时间作用。拟议的研究结果有望进一步加深我们对这些众多病毒粒子蛋白的作用以及它们相互作用对感染性颗粒形成的重要性的理解。为实现这些目标提出的具体目标是：具体目标1：保守的被盖基因的遗传相互作用网络：pUL21的关键作用。具体目标2：阐明以丰富的被膜蛋白VP22为中心的“合成致死”的表型，以确定导致病毒成熟和包膜的途径。

Herpesviruses are human pathogens that cause life-long persistent infections and result in clinical manifestations that range from a mild cold sore to cancer. Herpes simplex virus (HSV) is among the most frequently encountered virus by the general population. Infection with HSV-1 often results in orolabial disease but it can also cause ocular disease (keratitis) that can lead to blindness or encephalitis, which can be fatal. Type-2 infections are characterized by genital ulcerative disease and infections of the newborn. Thus, infections due to HSV are a major public health concern and understanding the biology of this virus is important in the development of efficacious treatments of these infections. The HSV-1 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 tegument is one of the most complex and diverse structures of the virion, both in terms of protein composition and the functions encoded by the constituents of this structure. Of this large and diverse group of virion proteins, not all are classified as being essential for replication in a laboratory cell culture system. What are the functions of all the “non-essential” proteins for virion maturation and egress? These “accessory” gene products likely play important roles in the infected host but an in-depth discovery of their functions has not been accomplished in a comprehensive manner. Using a genome assembly line coupled with synthetic genomics methods, we can elucidate the functions of these proteins by creating “synthetic lethality” in a mutant virus. Our working hypothesis posits that by phenotypic investigation of synthetic lethals, we can then begin to formulate pathways in which these “redundant” virion proteins participate, as well as, their spatial/temporal roles during HSV-1 virion morphogenesis. The outcome of the proposed studies is expected to further our understanding of the roles of these multitude virion proteins and the importance of their interactions for infectious particle formation. The Specific Aims proposed to achieve these goals are: Specific Aim 1: Genetic interaction networks of the conserved tegument genes: Pivotal role of pUL21. Specific Aim 2: Elucidate the phenotypes of “synthetic lethals” that are centered around the abundant tegument protein, VP22 to determine pathways that lead to virus maturation and envelopment.