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.

疱疹病毒是人类病原体，其引起终身持续感染，并导致从轻度唇疱疹到癌症的临床表现。单纯疱疹病毒（HSV）是最常见的病毒之一。HSV-1感染通常会导致口唇疾病，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疡性疾病和新生儿感染。因此，HSV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非常重要。HSV-1病毒粒子由四个结构元件组成：位于中心核心的大的双链DNA分子;包围基因组的二十面体衣壳;直接包围衣壳的层，称为被膜;以及外膜或包膜，其包围整个结构并在其中嵌入病毒糖蛋白。被膜是病毒体最复杂和多样的结构之一，在蛋白质组成和由该结构的成分编码的功能方面。在这一庞大而多样的病毒体蛋白质组中，并非所有都被归类为在实验室细胞培养系统中复制所必需的。病毒粒子成熟和排出的所有“非必需”蛋白质的功能是什么？这些“附属”基因产物可能在受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。使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建“合成致死性”来阐明这些蛋白质的功能。我们的工作假设假定，通过对合成致死物的表型研究，我们可以开始制定这些“冗余”病毒体蛋白参与的途径，以及它们在HSV-1病毒体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病毒体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：皮层保守基因的遗传相互作用网络：pUL 21的功能。具体目标二：阐明以丰富的被膜蛋白VP 22为中心的“合成致死”的表型，以确定导致病毒成熟和死亡的途径。