CAREER: The Lassa Virus Fusion Domain - A Cyclic Approach to Revealing the Molecular Chemistry behind Membrane Fusion

职业：拉沙病毒融合域 - 揭示膜融合背后分子化学的循环方法

• 批准号: 2238139
• 负责人: Jin Woo Lee
• 金额: $ 82万
• 依托单位: University of Maryland, College Park
• 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: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2238139&HistoricalAwards=false
• 关键词: CAREER; Lassa; Virus; Fusion; Domain

With the support of the CLP program in the Division of Chemistry, Jin Woo Lee from the University of Maryland, College Park is studying the molecular chemistry behind the initiation of Lassa Virus (LASV) membrane fusion. LASV is a member of the Arenavirus family that is currently endemic to West Africa, and there is an increasing incidence of LASV infection reported worldwide. The pandemic potential of LASV has already been noted by the World Health Organization (WHO) in 2018, which stated that the virus requires urgent and prioritized research. Membrane fusion is a key component of the LASV lifecycle that allows the virus to deliver its genetic information into the target cell through a chemical process that is not well understood. The process is thought to be facilitated by specialized fusion machinery that undergoes structural transitions triggered by specific environmental conditions. One such example of this machinery is the fusion domain (FD), which is predominately associated with the initiation of membrane fusion. The research aims to identify, on a molecular level, exactly how the FD initiates the process of membrane fusion. Such information is often a prerequisite for novel therapeutic design that may serve to arrest the viral lifecycle and prevent and/or treat LASV infection. Simultaneously, an educational outreach program will be implemented focused on providing opportunities for incoming transfer students as well as local high school students to gain practical scientific research experience. The aim of the program is to improve both the retention and enrollment of students within the science, technology, engineering, and mathematics (STEM) fields.The initiation of LASV membrane fusion relies on the FD, which resides within the viral glycoprotein found on the viral surface. This domain is well conserved across the Arenavirus family and is also structurally distinct, containing both an N-terminal fusion peptide (FP) and an internal fusion loop (FL). The overarching goal of the research is to understand how the underlying chemistry of the FD contributes to the initiation of membrane fusion. Specifically, the structural and functional roles of multiple fusogenic regions within the LASV FD and the impacts of environmental conditions will be examined. A cyclic approach will be employed utilizing solution nuclear magnetic resonance (NMR) spectroscopy to elucidate the structural rearrangement of the LASV FD. The NMR work will be complemented by thermodynamic and kinetic investigations through isothermal titration calorimetry (ITC) and in vitro lipid mixing fusion assays. The framework mentioned allows the investigation of the FD as both a peptide and within the context of the full viral glycoprotein. The latter requires a novel approach involving specific labeling and incorporation into a lipid bilayer, both of which will be implemented in order to provide more physiological relevance. Upon conclusion of the research, a major knowledge gap regarding LASV membrane fusion will be filled and a framework for studying other viral FDs will be established.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.

在化学系 CLP 项目的支持下，马里兰大学帕克分校的 Jin Woo Lee 正在研究拉沙病毒 (LASV) 膜融合启动背后的分子化学。 LASV 是沙粒病毒家族的成员，目前在西非流行，并且全球范围内报告的 LASV 感染发病率不断增加。世界卫生组织 (WHO) 已于 2018 年注意到 LASV 的大流行潜力，并表示需要对该病毒进行紧急和优先的研究。膜融合是 LASV 生命周期的关键组成部分，它允许病毒通过一种尚不清楚的化学过程将其遗传信息传递到靶细胞中。人们认为，这一过程是由专门的聚变机制促进的，这些机制会经历由特定环境条件引发的结构转变。这种机制的一个例子是融合域（FD），它主要与膜融合的启动相关。该研究旨在从分子水平上确定 FD 究竟如何启动膜融合过程。此类信息通常是新颖治疗设计的先决条件，可用于阻止病毒生命周期并预防和/或治疗 LASV 感染。同时，将实施一项教育推广计划，重点为即将入学的转学生和当地高中生提供获得实际科学研究经验的机会。该计划的目的是提高科学、技术、工程和数学 (STEM) 领域学生的保留率和入学率。LASV 膜融合的启动依赖于 FD，FD 存在于病毒表面的病毒糖蛋白内。该结构域在沙粒病毒家族中非常保守，并且结构也不同，包含 N 端融合肽 (FP) 和内部融合环 (FL)。该研究的首要目标是了解 FD 的基本化学成分如何促进膜融合的启动。具体来说，将检查 LASV FD 内多个融合区域的结构和功能作用以及环境条件的影响。将采用循环方法，利用溶液核磁共振 (NMR) 光谱来阐明 LASV FD 的结构重排。 NMR 工作将得到通过等温滴定量热法 (ITC) 和体外脂质混合融合测定的热力学和动力学研究的补充。所提到的框架允许对 FD 作为肽和完整病毒糖蛋白进行研究。后者需要一种涉及特异性标记和掺入脂质双层的新方法，这两者都将被实施以提供更多的生理相关性。研究结束后，将填补有关 LASV 膜融合的主要知识空白，并将建立研究其他病毒 FD 的框架。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

A Salt Bridge and Disulfide Bond within the Lassa Virus Fusion Domain Are Required for the Initiation of Membrane Fusion

• DOI: 10.1021/acsomega.3c08632
• 发表时间: 2024-01-18
• 期刊: ACS OMEGA
• 影响因子: 4.1
• 作者: Pennington, Hallie N.;Birtles, Daniel;Shi, Zoe W.;Lee, Jinwoo
• 通讯作者: Lee, Jinwoo

SARS-CoV-2 Fusion Domain Provides Clues toward the Molecular Mechanism for Membrane Fusion

SARS-CoV-2融合域为膜融合的分子机制提供线索

• DOI: 10.1021/acs.biochem.3c00501
• 发表时间: 2023
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Birtles, Daniel;Lee, Jinwoo
• 通讯作者: Lee, Jinwoo