MCA: Using Multiple Approaches for Understanding RNA Binding by Enzymes in Intermediary Metabolism

MCA：使用多种方法了解中间代谢中酶与 RNA 的结合

• 批准号: 2321442
• 负责人: Constance Jeffery
• 金额: $ 41.42万
• 依托单位: University of Illinois at Chicago
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2321442&HistoricalAwards=false
• 关键词: MCA; Using; Multiple; Approaches; Understanding

All cells need methods to adjust their activities in a coordinated fashion to respond to changes in their environment. In recent years, dozens of proteins that have central roles in obtaining energy from food have, surprisingly, been found to have direct interactions with specific RNA molecules that are key players in turning up or down other cellular activities. A greater understanding of how these proteins interact with and control RNA functions may lead to the design and development of novel methods to regulate many kinds of cellular activities, as well as commercial biotechnology applications. This Mid-Career Advancement (MCA) project will enable the principal investigator to receive hands-on training in determining the structures of protein/RNA complexes at a national cryo-electron microscopy research facility. The PI will also work with an expert collaborator to gain experience in using computational methods for analysis and prediction of protein/RNA interactions. The project will generate opportunities for undergraduate students to gain research experience and with it the mentoring and training needed to prepare them for future careers in science. Training of individuals with these skills is important to the high-tech industries in the United States. Dynamic interactions between RNA and RBPs play key roles in all aspects of post-transcriptional gene regulation, including splicing, transport, translation, and maintaining RNA stability or promoting RNA degradation. Combining catalytic and RNA binding functions in one multifunctional protein can be a mechanism to coordinate cellular activities, for example, by sensing the cell’s metabolic state through availability of the enzyme’s ligands and responding by regulating translation of specific transcripts. Conversely, RNA binding could regulate the enzyme’s catalytic activity. Cryo-electron microscopy for determining the structures of enzyme/RNA complexes will complement computational methods for analysis and prediction of RNA binding sites on proteins. Structure-guided mutagenesis will be used for validation of RNA/protein interaction sites. Successful completion of the proposed project will add to the limited number of structures of complexes containing RNA bound to noncanonical RNA binding proteins and provide valuable information about the mechanisms of RNA binding. The computer-based analysis will aid in understanding the nature of the interacting surfaces and will provide a basis for identifying other unconventional RNA binding proteins. This information can be applied in the future to the design and development of novel proteins that regulate RNA translation, stability, and lifetime and RNAs that regulate enzyme function.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.

所有细胞都需要方法以协调的方式调整其活动，以响应环境的变化。 近年来，令人惊讶的是，人们发现数十种在从食物中获取能量方面发挥核心作用的蛋白质与特定的 RNA 分子有直接相互作用，而这些分子是调节其他细胞活动的关键因素。更好地了解这些蛋白质如何与 RNA 相互作用并控制 RNA 功能可能会导致设计和开发调节多种细胞活动的新方法以及商业生物技术应用。该中期职业发展 (MCA) 项目将使主要研究人员能够在国家冷冻电子显微镜研究设施中接受确定蛋白质/RNA 复合物结构的实践培训。 PI 还将与专家合作者合作，获得使用计算方法分析和预测蛋白质/RNA 相互作用的经验。该项目将为本科生提供获得研究经验的机会，并为他们未来的科学职业做好准备所需的指导和培训。培训具有这些技能的个人对于美国的高科技产业非常重要。 RNA 和 RBP 之间的动态相互作用在转录后基因调控的各个方面发挥着关键作用，包括剪接、运输、翻译以及维持 RNA 稳定性或促进 RNA 降解。将催化和 RNA 结合功能结合在一个多功能蛋白中可以成为协调细胞活动的机制，例如，通过酶配体的可用性来感知细胞的代谢状态，并通过调节特定转录本的翻译来做出响应。相反，RNA 结合可以调节酶的催化活性。用于确定酶/RNA复合物结构的冷冻电子显微镜将补充用于分析和预测蛋白质上RNA结合位点的计算方法。结构引导诱变将用于验证 RNA/蛋白质相互作用位点。该项目的成功完成将增加有限数量的含有与非规范RNA结合蛋白结合的RNA的复合物结构，并提供有关RNA结合机制的有价值的信息。基于计算机的分析将有助于理解相互作用表面的性质，并为识别其他非常规 RNA 结合蛋白提供基础。这些信息将来可以应用于调节 RNA 翻译、稳定性和寿命的新型蛋白质以及调节酶功能的 RNA 的设计和开发。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。