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.

疱疹病毒是引起终生持续感染并导致从轻度唇疱疹到癌症的临床表现的人类病原体，单纯疱疹病毒 (HSV) 是普通人群最常见的 HSV-1 感染病毒之一。导致口唇疾病，但也可引起眼部疾病（角膜炎），从而导致失明或脑炎，2 型感染的特征是生殖器溃疡病和新生儿感染。因此，HSV 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。HSV-1 病毒体由四个结构元件组成：一个大的双链 DNA 分子。在中央核心；一个二十面体衣壳，它包围着基因组；以及一个外膜或包膜，它包围着整个结构，其中嵌入病毒糖蛋白 被膜是病毒体中最复杂和最多样化的结构之一，无论是从蛋白质组成还是由该结构的成分编码的功能来看，在这一庞大而多样化的病毒体蛋白质组中，并非所有病毒体蛋白质都被分类。对于实验室细胞培养系统中的复制至关重要，这些“辅助”基因产物可能在受感染宿主中发挥重要作用，但对病毒颗粒的成熟和排出具有哪些重要作用？发现使用基因组组装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产生“合成致死性”来阐明这些蛋白质的功能，我们的工作假设是通过表型研究来阐明它们的功能。的合成致死，然后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。预计将进一步加深我们对这些众多病毒体蛋白的作用及其相互作用对于感染性颗粒形成的重要性的理解。 为实现这些目标而提出的具体目标是： 具体目标 1：保守外皮基因的遗传相互作用网络：关键作用。 pUL21 的具体目标 2：阐明以丰富的外皮蛋白 VP22 为中心的“合成致死细胞”的表型，以确定导致病毒成熟和传播的途径。包围。