RAPID: Comparative functional characterization of strain-specific CoV E-proteins and involvement in host-specific virulence

在过去的几十年里，不同毒株的冠状病毒感染导致了SARS-CoV(2003年)和MERS(2012年)的流行，以及最近的新冠肺炎(2019年)大流行。在这一点上，还没有针对这些人类病原体的疫苗或有效的抗病毒药物。冠状病毒由四种主要结构蛋白组成：吞噬病毒核糖核酸的核衣壳蛋白(N)、刺突蛋白(S)、膜蛋白(M)和包膜蛋白(E)。目前，学术和药物方面的努力主要集中在S蛋白上，该蛋白参与病毒进入宿主细胞，而M和E蛋白研究较少，但被证明参与病毒复制。本项目将对不同冠状病毒株的E蛋白进行研究，以更好地了解其功能。这个项目将提高我们对SARS-CoV-2的了解，这种了解是确定控制当前Covi-19大流行的新疗法所必需的知识。除了增加关于SARS-CoV-2生物学的知识外，该项目还支持对一名博士后研究员的培训，扩大对STEM的参与。本项目将研究SARS-CoV-2和其他冠状病毒(CoV)的E-蛋白，以了解该离子通道在宿主-病毒粒子相互作用中的作用。将使用多水平的方法来全面了解来自不同冠状病毒株的各种E蛋白的翻译后修饰和序列变异的影响。将确定E蛋白翻译后修饰的模式，以确定这些修饰是否影响模型膜中E蛋白的功能。使用模型膜可以控制膜环境中的关键特征，即脂头基团的电荷和脂肪酸的饱和度，从而创建微域或非微域环境。通过使用我们将现代电子学和膜生物物理学相结合的独特能力，这些记录的吞吐量将会增加。来自不同冠状病毒株的E蛋白将被用来确定在类似宿主的环境中对生产性或非生产性感染发展的影响。当前的新冠肺炎突发公共卫生事件使了解这种新病原体变得刻不容缓。人们对这种新型冠状病毒及其与其他冠状病毒粒子的关系知之甚少。更好地了解E蛋白在宿主-病毒粒子相互作用中的确切作用将有助于设计新的治疗方法。这一快速奖项是由综合组织系统生物部的生理和结构系统集群利用冠状病毒援助、救济和经济安全(CARE)法案的资金颁发的。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Viral infections in the last decades with different strains of the coronavirus (CoV) have led to the SARS-CoV (2003) and MERS (2012) epidemics and to the most recent COVID-19 pandemic (2019). At this point, no vaccine or effective antiviral drug is commercially available for these human pathogens. CoVs are built out of four major structural proteins; the nucleocapsid (N) protein engulfing the viral RNA, the spike (S) protein, the membrane (M) protein and the envelope (E) protein. Presently, academic and pharmaceutical efforts are mainly focused on the S-protein which is involved in the entry of the virus into the host cell, whereas the M- and E-proteins are less well studied but are shown to be involved in viral replication. This project will study the E-protein from different CoV strains to better understand its function. This project will improve our understanding of SARS-CoV-2, and this understanding is knowledge necessary to identify new therapeutics to control this current COVI-19 pandemic. In addition to increasing knowledge about SARS-CoV-2 biology, this project also supports the training of a post-doctoral fellow, broadening participation in STEM. This project will study the E-protein of SARS-CoV-2 and other coronaviruses (CoVs) to understand the role of this ion channel in host-virion interaction. A multi-level approach will be used to gain comprehensive understanding on effects of the post-translational modifications and sequence variability of various E-proteins from different CoV strains. Patterns in the post-translational modification of E-proteins will be identified to determine whether these modifications affect the functionality of the E-protein in model membranes. Using model membranes allows for control of key-features in the membrane environment, namely charge of lipid head-groups and saturation of fatty acids creating a microdomain or non-microdomain environment. The throughput of these recordings will be increased by using our unique abilities to combine modern electronics and membrane biophysics. E-proteins from different CoV strains will be used to identify the impact on the development of productive or non-productive infections in a host-like environment. The current COVID-19 public health emergency makes understanding this new pathogen of urgent importance. Little is known about this novel CoV and its relationship to other CoV virions. Understanding better the exact role that the E-protein plays in the host-virion interaction will be very helpful in devising new therapeutics. This RAPID award is made by the Physiological and Structural Systems Cluster in the BIO Division of Integrative Organismal Systems, using funds from the Coronavirus Aid, Relief, and Economic Security (CARES) Act.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.