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.

所有细胞都需要以协调的方式调整其活动的方法，以应对其环境变化。近年来，令人惊讶的是，数十种在从食物中获取能量中具有主要作用的蛋白质与特定的RNA分子有直接相互作用，这些蛋白质与特定的RNA分子有直接相互作用，这些分子是推动其他细胞活动的关键参与者。对这些蛋白质如何与这些蛋白质相互作用和控制RNA功能的更深入了解可能会导致新方法的设计和开发，以调节许多类型的细胞活性以及商业生物技术应用。这个职业中期进步（MCA）项目将使首席研究人员能够接受动手培训，以确定国家冷冻电子显微镜研究设施的蛋白质/RNA复合物的结构。 PI还将与专家合作者合作，以获得使用计算方法来分析和预测蛋白质/RNA相互作用的经验。该项目将为本科生获得获得研究经验的机会，并借助它为未来的科学职业做好准备所需的心理和培训。对具有这些技能的个人的培训对于美国的高科技行业很重要。 RNA和RBP之间的动态相互作用在转录后基因调节的各个方面起关键作用，包括剪接，传输，翻译和维持RNA稳定性或促进RNA降解。在一种多功能蛋白中结合催化和RNA结合功能可以是一种可以通过酶的配体可用来传感细胞的代谢状态来协调细胞活性的机制，并通过调节特定转录本的翻译来响应。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RNA结合位点的计算方法。结构引导的诱变将用于验证RNA/蛋白质相互作用位点。拟议项目的成功完成将增加包含与非规范RNA结合蛋白结合的RNA的复合物的有限数量，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。将来可以将这些信息应用于调节调节酶功能的RNA翻译，稳定性以及寿命和RNA的新型蛋白质的设计和开发。该奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和更广泛的影响审查标准来通过评估来通过评估来支持的。