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

疱疹病毒是一种人类病原体，可引起终身持续性感染，其临床表现从轻微的唇疱疹到癌症不等。单纯疱疹病毒（HSV）是普通人群中最常见的病毒之一。感染1型单纯疱疹病毒通常会导致口唇病，但它也可能导致眼部疾病（角膜炎），从而导致失明或脑炎，这可能是致命的。2型感染的特点是生殖器溃疡和新生儿感染。因此，由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特性对于开发有效治疗这些感染的方法非常重要。HSV-1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。被膜是病毒粒子最复杂和最多样化的结构之一，无论是蛋白质组成还是由该结构成分编码的功能。在这一庞大而多样的病毒粒子蛋白质群中，并非所有的病毒粒子蛋白质都被归类为实验室细胞培养系统中复制所必需的。对于病毒粒子的成熟和输出，所有“非必需”蛋白质的功能是什么？这些“附属”基因产物可能在被感染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。利用基因组装配线结合合成基因组学方法，我们可以通过在突变病毒中创造“合成致死性”来阐明这些蛋白质的功能。我们的工作假设假设，通过对合成致死病毒的表型研究，我们可以开始制定这些“冗余”病毒粒子蛋白参与的途径，以及它们在HSV-1病毒粒子形态发生过程中的空间/时间作用。这些研究的结果有望进一步加深我们对这些多种病毒粒子蛋白的作用及其相互作用对感染性颗粒形成的重要性的理解。具体目标1：保守被膜基因的遗传相互作用网络：pUL21的关键作用。特异性目标2：阐明以丰富的被膜蛋白VP22为中心的“合成致死病毒”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.