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

阐明 HSV-1 病毒体蛋白的遗传网络并发现它们在感染性病毒颗粒形态发生中的功能

• 批准号: 10319969
• 负责人: PRASHANT J DESAI
• 金额: $ 64.67万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-10 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10319969
• 关键词: Biochemical; Biological; Biological Assay; Biology; Blindness; Capsid; Cell Culture System; Cell Culture Techniques; Cells; Clinical; Complex; Coupled; DNA Packaging; Data; Defect; Development; Disease; Dissection; Electrons; Elements; Encephalitis; Engineering; Environment; Family; Gene Deletion; General Population; Genes; Genetic; Genital; Genitalia; Genome; Glycoproteins; Goals; Herpes Labialis; Herpesviridae; Herpesvirus 1; Individual; Infection; Invaded; Investigation; Keratitis; Laboratories; Lead; Life; Malignant Neoplasms; Mediating; Membrane; Membrane Proteins; Methods; Molecular Genetics; Morphogenesis; Mutation; Mutation Analysis; Nuclear; Outcome; Pathway interactions; Phenotype; Play; Proteins; Public Health; Publishing; Role; Side; Simplexvirus; Structure; Testing; Ulcer; Viral; Virion; Virus; base; cellular imaging; chronic infection; combinatorial; ds-DNA; efficacious treatment; experience; gene network; gene product; genome-wide; human pathogen; imaging modality; insight; mutant; neonatal infection; novel; particle; pathogen; polypeptide; protein complex; protein function; synthetic biology; synthetic genomics; tool; viral DNA

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.

疱疹病毒是引起终身持续感染的人类病原体，并导致临床表现，从轻度的冷疮到癌症。单纯疱疹病毒（HSV）是普通人群最常遇到的病毒之一。 HSV-1感染通常会导致口腔疾病，但也可能引起眼部疾病（角膜炎），可能导致失明或脑炎，这可能是致命的。 2型感染的特征是生殖器性溃疡性疾病和新生儿的感染。这是HSV引起的感染是一个主要的公共卫生关注，并且了解该病毒的生物学在开发这些感染的有效治疗方面很重要。 HSV-1病毒完成了四个结构元素：中心核中的大型双链DNA分子；一个包围基因组的二十面体capsid；一个立即周围环境的层被称为tegument；和外膜或包膜，它包围了整个结构，并且嵌入了病毒糖蛋白的嵌入，团队是病毒粒子最复杂和最不同的结构之一，无论是在蛋白质组成方面还是由该结构的构建体编码的功能。在这种大而不同的病毒蛋白蛋白中，并非所有人都被归类为在实验室细胞培养系统中复制至关重要。所有“非必需”蛋白的功能是病毒成熟和出口的功能是什么？这些“附件”基因产品可能在感染的宿主中起重要作用，但是深入发现其功能并未以全面的方式完成。使用与合成基因组方法结合的基因组组装线，我们可以通过在突变病毒中创建“合成的致死性”来阐明这些蛋白质的功能。我们的工作假设表明，通过对合成致命的表型投资，我们可以开始构建这些“冗余”病毒素蛋白参与的途径，以及它们在HSV-1毒素形态发生过程中的空间/时间角色。预期拟议的研究的结果将进一步了解这些众多病毒蛋白蛋白的作用以及它们相互作用对传染性颗粒形成的重要性。提出实现这些目标的具体目的是：特定目标1：配置的TEGUMENT基因的遗传相互作用网络：PUL21的关键作用。具体目的2：阐明围绕绝对tegument蛋白（VP22）的“合成致死性”的表型，以确定导致病毒成熟和环境的途径。