RAPID: Impact of SARS-CoV2 on function of the cellular tRNA methyltransferase 1 and redox homeostasis

由于SARS-CoV2引起的全球大流行，迫切需要了解冠状病毒感染是如何影响宿主细胞过程的。该项目将探索最近发现的SARS-CoV2蛋白裂解酶与可能成为其目标的人类蛋白之间的联系。将使用生化、分子和细胞相结合的方法来研究病毒和人类蛋白质的相互作用如何改变正常的细胞功能。预计结果将揭示应对新冠肺炎大流行的治疗干预目标。该项目将为开展这项研究的两名研究生提供培训机会。最近，在SARS-CoV2的主要蛋白酶Nsp5和人类tRNA甲基转移酶TRMT1之间发现了一种新的联系。TRMT1在超过一半的细胞tRNA和许多线粒体tRNA中甲基化一个特定的位置(m2.2G)，已知TRMT1是人类细胞正常水平的蛋白质合成和氧化还原动态平衡所必需的。TRMT1含有一个氨基酸序列，可能是Nsp5切割的目标，但Nsp5-TRMT1相互作用的生物学效应和下游后果尚不清楚。本项目将集中于三个目标：确定NSP5结合对TMRT1‘S稳定性和结合tRNAs能力的影响；识别通过NSP5与TRMT1相互作用而调节的依赖TRMT1的通路；以及测试依赖TRMT1的通路如何影响SARS-CoV2的生命周期。通过了解SARS-CoV2感染如何影响TRMT1功能，该项目将深入了解病毒用来改变宿主基因表达和新陈代谢的新策略。这一结果可能有助于开发新的药物疗法或检测冠状病毒感染的生物标记物。这一快速奖项是由分子和细胞生物科学部门的遗传机制计划利用冠状病毒援助、救济和经济安全(CARE)法案的资金做出的。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。

Due to the global pandemic caused by SARS-CoV2, there is an urgent need to understand how coronavirus infection affects host cellular processes. This project will explore a recently discovered link between a SARS-CoV2 protein-cleaving enzyme and a human protein that may be its target. Combined biochemical, molecular, and cellular approaches will be used to study how interaction of the viral and human proteins can alter normal cellular functions. The outcomes are expected to uncover targets for therapeutic intervention to address the COVID-19 pandemic. The project will provide training opportunities for two graduate students who will carry out the research. Recently, a novel connection was discovered between the main protease of SARS-CoV2, called Nsp5, and the human tRNA methyltransferase enzyme known as TRMT1. TRMT1 methylates a specific position (m2.2G) in more than half of all cellular tRNAs and in many mitochondrial tRNAs, and TRMT1 is known to be essential for normal levels of protein synthesis and for redox homeostasis in human cells. TRMT1 contains an amino acid sequence that may be a target for cleavage by Nsp5, but the biological effects and downstream consequences of the Nsp5-TRMT1 interaction are unknown. This project will focus on three aims: determining the effects of Nsp5 binding on TMRT1's stability and ability to bind tRNAs; identifying TRMT1-dependent pathways that are modulated through Nsp5 interaction with TRMT1; and testing how TRMT1-dependent pathways impact the SARS-CoV2 life cycle. By understanding how SARS-CoV2 infection affects TRMT1 function, the project will yield insight into novel strategies that viruses employ to alter host gene expression and metabolism. The results could aid in development of new drug therapies or biomarkers for detecting coronavirus infection. This RAPID award is made by the Genetic Mechanisms Program in the Division of Molecular and Cellular Biosciences, 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.