Identifying structural features of viral RNA recognized by host helicases

鉴定宿主解旋酶识别的病毒 RNA 的结构特征

• 批准号: RGPIN-2017-04003
• 负责人: Patel, Trushar
• 金额: $ 2.26万
• 依托单位: University of Lethbridge
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=661701
• 关键词: Identifying; structural; features; viral; RNA

Viruses hijack the host cell machinery and recruit host proteins to aid viral replication. Considerable effort has been made to identify host proteins that interact with viral RNA/proteins. However, the biochemical and structural basis of viral RNA recognition by the members of the DEAD-box family, which support viral replication, is poorly understood. My long-term research goal is to study these key host helicases that are exploited by many viruses in order to gain detailed insights into the molecular mechanisms behind a broad spectrum of host helicases and viral RNA interactions. Despite the ATP-dependent unwinding of nucleic acids being the established role of DEAD-box helicases, recent studies demonstrate that helicases influence all major aspects of RNA metabolism and often play significant roles in viral replication. Particularly, the helicases DDX3X and DDX5 interact with the viral 3' untranslated region (3' UTR) of Japanese encephalitis virus, hepatitis C virus, and SARS coronavirus, and support their replication. My laboratory will investigate the structural basis of the interactions of DDX3X/DDX5 with 3' UTRs of members of the Flavivirus genus (e.g., Japanese encephalitis, West Nile, Yellow Fever, Zika, and Dengue viruses) to identify specific patterns of viral RNA being recognized by the helicases under investigation. An interdisciplinary approach involving molecular biology, biochemistry, biophysical and structural biology techniques will be employed to achieve these goals. In the longer term, my research program will consider other members of the helicase family (e.g., DDX1, DDX6, and DDX17), as well as viruses that infect plants (e.g., tomato bushy stunt virus) to establish if the recognition of viral 3' UTR is a conserved function of DEAD-box helicases across the species. In addition, studying helicase-viral RNA and helicase-host RNA interactions will allow insights into how helicases discriminate viral and host RNAs. My research program will provide a platform for training highly qualified personnel in a multidisciplinary research area using a hands-on approach and cutting-edge techniques (e.g., light scattering, analytical ultracentrifuge, small angle X-ray scattering and X-ray crystallography).

病毒劫持宿主细胞机器并募集宿主蛋白质以帮助病毒复制。已经做出相当大的努力来鉴定与病毒RNA/蛋白质相互作用的宿主蛋白质。然而，支持病毒复制的DEAD盒家族成员识别病毒RNA的生化和结构基础知之甚少。我的长期研究目标是研究这些被许多病毒利用的关键宿主解旋酶，以便详细了解广泛的宿主解旋酶和病毒RNA相互作用背后的分子机制。尽管ATP依赖的核酸解旋是DEAD盒解旋酶的既定作用，但最近的研究表明，解旋酶影响RNA代谢的所有主要方面，并且通常在病毒复制中起重要作用。特别地，解旋酶DDX 3X和DDX 5与日本脑炎病毒、丙型肝炎病毒和SARS冠状病毒的病毒3'非翻译区（3' UTR）相互作用，并支持它们的复制。我的实验室将研究DDX 3X/DDX 5与黄病毒属成员的3'UTR相互作用的结构基础（例如，日本脑炎、西尼罗河、黄热病、寨卡和登革热病毒），以鉴定被研究中的解旋酶识别的病毒RNA的特定模式。将采用涉及分子生物学、生物化学、生物物理学和结构生物学技术的跨学科方法来实现这些目标。从长远来看，我的研究计划将考虑解旋酶家族的其他成员（例如，DDX 1、DDX 6和DDX 17），以及感染植物的病毒（例如，番茄丛矮病毒）以确定病毒3' UTR的识别是否是跨物种的DEAD盒解旋酶的保守功能。此外，研究解旋酶-病毒RNA和解旋酶-宿主RNA的相互作用将有助于深入了解解旋酶如何区分病毒和宿主RNA。我的研究计划将提供一个平台，培养高素质的人才在多学科研究领域使用动手的方法和尖端技术（例如，光散射、分析超散射、小角X射线散射和X射线晶体学）。