Dissecting the mechanism of herpesvirus genome packaging

剖析疱疹病毒基因组包装机制

• 批准号: 10509325
• 负责人: Allison Louise Didychuk
• 金额: $ 50.25万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-25 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10509325
• 关键词: Address; Adopted; Africa South of the Sahara; Antiviral Agents; Area; Award; Bacteriophages; Behavior; Biochemical; Biochemistry; Biological Assay; Biology; Biophysics; Cancer Etiology; Capsid; Charge; Complex; Congenital Abnormality; Coupled; DNA Viruses; Development; Disease; Dissection; Drug Targeting; Evolution; FDA approved; Foundations; Genome; Goals; HIV; Herpes Labialis; Herpes encephalitis; Herpesviridae; High Prevalence; Homologous Gene; Human; Human Herpesvirus 8; Immunocompromised Host; In Vitro; Individual; Infection; Investments; Kaposi Sarcoma; Label; Lead; Life; Malignant Neoplasms; Mass Spectrum Analysis; Methodology; Methods; Microscopy; Molecular; Molecular Motors; Molecular Virology; Monitor; Motor; Mutagenesis; Nucleic Acid Binding; Oncogenic; Pharmaceutical Preparations; Positioning Attribute; Process; Proteins; Proteomics; Regulation; Research Personnel; Resources; Role; Structure; System; Tail; Techniques; Testing; Time; Viral; Viral Genome; Viral Packaging; Viral Proteins; Virus; Virus Replication; Vision; Work; base; biophysical properties; drug discovery; high reward; high risk; human pathogen; in vivo; inhibitor; innovation; insight; interest; mutation screening; neonate; new therapeutic target; novel; pathogen; programs; reconstitution; resistance mutation; scaffold; structural biology; success; terminase; virology

PROJECT SUMMARY Human herpesviruses can cause congenital birth defects in neonates and a wide range of disease in humans, with particularly devastating effects in immunocompromised individuals. Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of Kaposi's sarcoma and other cancers, with high prevalence in sub- Saharan Africa. There is no cure for any herpesvirus; most existing antivirals target a single viral process and can lead to resistance mutations. The recent development of a herpesviral packaging inhibitor highlights the viral genome packaging machinery as an attractive target for novel antivirals. The herpesviral packaging machinery is conceptually similar to that of bacteriophages, yet it has additional complexities that include factors of unknown function. In this proposal, I outline my vision to understand the molecular mechanism of herpesvirus packaging. I will determine the function of an essential factor in herpesvirus packaging that has long evaded mechanistic dissection using state-of-the-art proteomics and microscopy approaches. I will also develop a new strategy to apply deep mutational scanning approaches to herpesviral proteins. In parallel, I will determine the structure of the KSHV terminase, the viral molecular motor that packages the viral genome into the nascent capsid. I will then reconstitute the essential components of the KSHV packaging machinery and use these to develop an in vitro packaging assay. This assay will be leveraged to screen for new antivirals and determine the molecular underpinnings of existing drugs that target the packaging machinery. The proposed studies will build a foundation to address long-standing, fundamental questions in DNA virus packaging and understand the mechanism of anti-herpesvirals.

项目摘要 人类疱疹病毒可能会导致新生儿的先天性先天性缺陷和人类多种疾病， 在免疫功能低下的个体中具有特别破坏性的影响。卡波西与肉瘤相关 疱疹病毒（KSHV）是Kaposi肉瘤和其他癌症的致病药物，在子 - 撒哈拉非洲。无法治愈任何疱疹病毒。大多数现有的抗病毒药针对单个病毒过程，并且 可能导致抗性突变。疱疹病毒包装抑制剂的最新发展突出了 病毒基因组包装机制是新型抗病毒药的有吸引力的目标。疱疹病毒包装 机械在概念上与噬菌体相似，但它具有其他复杂性，包括 功能未知因素。在此提案中，我概述了理解分子机制的愿景 疱疹病毒包装。我将确定具有疱疹病毒包装中重要因素的功能 长期使用最先进的蛋白质组学和显微镜方法逃避的机械剖析。我也会 制定一种新策略，将深层突变扫描方法应用于疱疹病毒蛋白。同时，我会 确定KSHV末端酶的结构，这是将病毒基因组包装到的病毒分子运动 新生的衣壳。然后，我将重建KSHV包装机械的基本组件，并 使用这些来开发体外包装测定法。该测定法将用于筛选新的抗病毒药和 确定针对包装机械的现有药物的分子基础。提议 研究将建立一个基础，以解决长期存在的DNA病毒包装中的基本问题和 了解抗赫普病毒的机制。