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.

项目总结 人类疱疹病毒会导致新生儿先天性出生缺陷和人类的多种疾病， 对免疫功能受损的人有特别毁灭性的影响。卡波西肉瘤相关 疱疹病毒(KSHV)是卡波西氏肉瘤和其他癌症的病原体，在亚裔中发病率很高。 撒哈拉非洲。任何疱疹病毒都无法治愈；大多数现有的抗病毒药物针对的是单一的病毒过程和 会导致抗药性突变。疱疹病毒包装抑制剂的最新发展突显了 病毒基因组包装机械作为新型抗病毒药物的有吸引力的目标。疱疹病毒包装 机械在概念上类似于噬菌体，但它有额外的复杂性，包括 未知函数的因子。在这项提议中，我概述了我的愿景，以了解 疱疹病毒包装。我将确定疱疹病毒包装中的一种基本因子的功能 长期以来，人们一直回避使用最先进的蛋白质组学和显微技术进行机械解剖。我也会 开发一种新的策略，将深度突变扫描方法应用于疱疹病毒蛋白。同时，我会 确定KSHV终止酶的结构，KSHV终止酶是病毒分子马达，将病毒基因组打包到 初露头角的衣冠。然后，我将重组KSHV包装机械的基本组件，并 利用这些来开发一种体外包装试验。这项检测将被用来筛选新的抗病毒药物和 确定针对包装机械的现有药物的分子基础。建议数 研究将建立一个基础，以解决长期存在的DNA病毒包装和 了解抗疱疹病毒药物的作用机制。