CAREER: Structural and mechanistic studies of RNA-mediated enteroviral genome replication

职业：RNA介导的肠道病毒基因组复制的结构和机制研究

• 批准号: 2236996
• 负责人: Deepak Koirala
• 金额: $ 78.57万
• 依托单位: University of Maryland Baltimore County
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-15 至 2028-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2236996&HistoricalAwards=false
• 关键词: CAREER; Structural; mechanistic; studies; RNA

Enteroviruses have a single-strand (+)-sense RNA genome containing complex RNA elements that direct genome replication by recruiting viral and host proteins to form ribonucleoprotein (RNP) complexes required for viral replication. Knowledge of the 3D structures of these RNA elements and corresponding RNPs is essential for understanding fundamental virological processes. The project will investigate the structural and mechanistic underpinnings of RNA-mediated enteroviral genome replication and determine how RNA structures exploit viral and host proteins to promote replication during viral infections. The findings can inform future therapeutic or preventive strategies to combat pathogens that cause human diseases and global pandemics. The project will also provide opportunities for RNA-centric research to a diverse group of high school, undergraduate, and graduate students, encouraging them to pursue higher studies and careers in RNA biochemistry, structural biology, or related STEM fields.The research will utilize antigen-binding antibody fragments (Fabs) as chaperones for determining the crystal structures of enteroviral replication-linked RNAs and RNPs and use biochemistry and molecular biology tools to elucidate RNA-RNA, RNA-protein, and protein-protein interactions. The objectives are to 1) determine the crystal structures of 5′ cis-acting replication elements (CREs) from seven major enteroviral genotypes (enterovirus A-C and rhinovirus A-D), 2) elucidate the interactions of these 5′CREs with viral 3C, 3CD fusion, and human PCBP proteins, and 3) investigate the crystal structures of 3′CREs and their RNPs formed within the 3′-end of the (-)-sense RNA replication intermediate. The outcomes are expected to reveal structure-function relationships that will provide new insights into the replication mechanism of this major class of viral pathogens.This project is jointly funded by the Genetic Mechanisms and Molecular Biophysics programs of the Molecular and Cellular Biosciences Division in the Biological Sciences Directorate.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

肠病毒具有单链（+） - 含有复杂的RNA元素，通过募集病毒蛋白和宿主蛋白形成核糖核蛋白（RNP）复合物，这些元素重复基因组复制。对这些RNA元素和相应RNP的3D结构的了解对于理解基本病毒学过程至关重要。该项目将研究RNA介导的肠病毒基因组复制的结构和机械基础，并确定RNA结构如何利用病毒蛋白和宿主蛋白来促进病毒感染期间的复制。这些发现可以为未来的治疗或预防策略提供抗击，以打击导致人类疾病和全球大流行病的病原体。该项目还将为一个高中，本科生和研究生的潜水者组为以RNA为中心的研究提供机会，鼓励他们从事RNA生物化学，结构生物学或相关STEM领域的更高研究和职业。该研究将利用抗原结合抗体碎片（Fabs）作为链结构的抗原结构，并确定肠结构的结构和肠结构的链接，并将其用于肠道结构。以及分子生物学工具，以阐明RNA-RNA，RNA - 蛋白质和蛋白质 - 蛋白质相互作用。目的是1）确定从七种主要的肠病毒基因型（肠病毒A-C和Rhinovirus a-d）中的5'顺式复制元件（CRE）的晶体结构，2）阐明这些5'Cr与病毒3C，3CD融合的相互作用，以及在3CD融合中的相互作用，以及在3CBP蛋白中及其晶体中的晶体及其晶体的相互作用。 3'-末端（ - ） - 感官RNA复制中间体。预计结果将揭示结构功能关系，将为这一主要病毒病原体的复制机制提供新的见解。该项目由分子和细胞生物学家的分子和细胞生物学家的遗传机制和分子生物体物质计划共同资助，该项目在生物科学局的基础上都通过recournional ofdional ofder of Infirinit the Infirinition Internation Insportional of Deemtion deemtion deemtorial deemorial deemorial deemorial siperiation deemorial siperiation deemorial siperiation deemorial siperiation deemorial siperial siperiation。和更广泛的影响审查标准。