CAREER: Molecular level consequences of mRNA modification

细胞面临着在正确的时间准确合成正确数量的蛋白质的艰巨挑战。信使核糖核酸（mRNA）是蛋白质合成的蓝图，mRNA的化学修饰是细胞调节蛋白质产生的一种途径。尽管越来越多的人认识到它们的重要性，但关于mRNA修饰的身份、流行率和功能后果的基本问题仍有待回答。该项目将填补关键的知识空白，使mRNA修饰如何影响蛋白质合成的分子水平的理解。这样一个框架对于确定mRNA修饰对关键生物过程的贡献至关重要。这项研究为学生提供了培训机会，并与旨在提高公众生物化学素养和激励未来一代科学家的并行教育计划相结合。努力包括通过密歇根大学自然历史博物馆（UMMNH）开发和展示实践教育经验。mRNA修饰的开创性研究将个体化学修饰的位置映射到数千个RNA分子。然而，目前尚不清楚大多数修饰如何（或在许多情况下，是否）从根本上改变mRNA的功能特性，以改变所得蛋白质的表达和/或身份。该项目使用了一套全面的生物物理化学和基于细胞的实验来开发一个机械基础，以了解化学修饰对核糖体如何“读取”mRNA代码的影响。此外，将开发和实施质谱分析工具，以确定酵母mRNA和MS 2噬菌体RNA基因组中修饰的身份。直接和定量方法的综合应用研究mRNA修饰对翻译的功能影响是该项目的一个显着特点。在一起，这些研究将揭示分子-mRNA修饰对蛋白质合成水平的影响。该项目由生物科学理事会分子和细胞生物科学部的遗传机制计划和数学和物理科学理事会化学部的生命过程化学计划共同资助。该奖项反映了NSF的法定使命，并被认为值得支持通过使用基金会的知识价值和更广泛的影响审查标准进行评估。

Cells face the daunting challenge of accurately synthesizing the correct number of proteins at the right time. Messenger ribonucleic acids (mRNAs) serve as blueprints for protein synthesis, and chemical modification of mRNA presents one avenue for cells to regulate protein production. Despite a rapidly growing recognition of their importance, fundamental questions regarding the identity, prevalence and functional consequences of mRNA modifications remain to be answered. This project will fill critical knowledge gaps to enable a molecular level understanding of how mRNA modifications impact protein synthesis. Such a framework will be imperative for ascertaining the contributions of mRNA modifications to key biological processes. This research provides training opportunities for students and is combined with a concurrent education plan aimed at increasing biochemical literacy in the public and motivating a future generation of scientists. Efforts include the development and presentation of hands-on educational experiences through the University of Michigan Museum of Natural History (UMMNH).Pioneering studies of mRNA modifications mapped the locations of individual chemical modifications to thousands of RNA molecules. However, it remains unclear how (or, in many cases, if) most modifications fundamentally change the functional properties of mRNAs to alter the expression and/or identity of the resulting proteins. This project uses a comprehensive set of biophysical chemistry and cell-based experiments to develop a mechanistic foundation for understanding the influence of chemical modifications on how the ribosome ‘reads’ the mRNA code. Furthermore, mass- spectrometry tools will be developed and implemented to ascertain the identity of modifications in yeast mRNAs and the MS2 bacteriophage RNA genome. The integrated application of direct and quantitative approaches to study the functional impact of mRNA modifications on translation is a distinguishing feature of this project. Together, these studies will reveal the molecular-level consequences of mRNA modifications on protein synthesis.This project is jointly funded by the Genetic Mechanisms program of the Molecular and Cellular Biosciences Division in the Biological Sciences Directorate and the Chemistry of Life Processes program of the Chemistry Division in the Mathematical and Physical 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.