CAREER: Determining The Fundamental Rules of RNA Tertiary Contact Formation

该奖项的部分资金来自《2021年美国救援计划法案》(公共法律117-2)。结构核糖核酸(RNA)在细胞功能中发挥关键作用，是RNA病毒功能的基础，也是生物传感器、生物分子支架和合成生物合成途径调节器等新型人造机器的蓝图。RNA结构由离散的和模块化的RNA基序组成-螺旋、连接和环。功能RNA通常包含两个或更多RNA基序之间的长距离三级接触，这些RNA基序将RNA链锁定为功能所需的明确结构。本项目旨在了解RNA结构中三级接触是如何形成的。三级接触是RNA结构的粘合剂，使它们能够形成复杂的3D结构。这个项目将破译它们形成的规则，最终产生第一个第三次接触形成的预测模型。同时，该计划旨在通过让学生和公民科学家参与新颖的基于科学的视频游戏，让参与者直接为研究成果做出贡献，来教育下一代科学家和公众关于RNA折叠和结构的知识。这将需要一门关于使用科学视频游戏的生物分子分子设计的新课程，这将允许学生设计新的生物分子，并将在实验室进行实验测试。这一综合研究和教育计划的结果将在利用RNA结构构建下一代RNA生物传感器和治疗方面具有重要的长期应用。结构RNA折叠成复杂的3D构象，执行基本的生物过程。这些RNA通常包含两个或更多RNA基序之间的长距离三级接触，这些RNA基序将RNA链锁定为功能所需的明确结构。目前，还没有可靠的方法可以预测三次接触的形成。预测第三次接触是具有挑战性的，因为尚不清楚哪些基序将形成第三次接触，以及这些基序是否可能接近形成相互作用。本项目旨在开发第一个三次接触形成的预测模型。为了实现这一目标，PI将使用新颖的3D RNA设计和大规模多路生物化学分析来确定哪些基序可以形成三级接触；确定允许三级接触伙伴靠近进行相互作用的RNA元件的三维构象灵活性，并建立三级接触形成模型并利用它来提高核酶的活性。三级接触形成预测模型的发展将提高对RNA调控过程的结构机制的基本理解，如基因表达、蛋白质翻译和mRNA剪接。该项目由生物科学局分子和细胞生物科学部的分子生物物理学分部和既定的激励竞争研究计划(EPSCoR)共同资助。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。

This award is funded in part under the American Rescue Plan Act of 2021 (Public Law 117-2).Structured ribonucleic acids (RNAs) play critical roles in cellular functions, underlie the function of RNA viruses, and serve as blueprints for new artificial machines such as biosensors, biomolecular scaffolds, and regulators of synthetic biosynthesis pathways. RNA structure is composed of discrete and modular RNA motifs – helices, junctions, and loops. Functional RNAs often contain long-range tertiary contacts between two or more RNA motifs that lock RNA strands into well-defined structures required for function. This project aims to understand how tertiary contacts form in RNA structure. Tertiary contacts are the “glue” of RNA structure allowing them to form complex 3D structures. This project will decipher the rules of their formation, ultimately generating the first predictive model of tertiary contact formation. In conjunction, the program seeks to educate the next generation of scientists and the public in RNA folding and structure by engaging students and citizen scientists in novel science-based video games that allow participants to directly contribute to research outcomes. This will entail a new course on the molecular design of biomolecules that employ scientific videos games, which will allow students to design new biomolecules that will be experimentally tested in the lab. Findings from this integrated research and education program will have important long-term applications in harnessing RNA structure to build the next generation of RNA biosensors and therapeutics.Structured RNAs fold into complex 3D conformations that perform fundamental biological processes. These RNAs often contain long-range tertiary contacts between two or more RNA motifs that lock RNA strands into well-defined structures required for function. Currently, there is no method that can reliably predict tertiary contact formation. Predicting tertiary contacts is challenging as it is unknown which motifs will form tertiary contacts and whether it is possible for these motifs to be brought into proximity to form an interaction. This program seeks to develop the first predictive model of tertiary contact formation. Towards this goal the PI will use novel 3D RNA design and massively multiplexed biochemical assays to identify which motifs can form tertiary contacts; determine the 3D conformational flexibility of RNA elements that permit tertiary contact partners to be brought into proximity to interact, and build tertiary contact formation model and utilize it to improve the activity of a ribozyme. The development of a predictive model of tertiary contact formation will improve the fundamental understanding of structural mechanisms of RNA-regulated processes such as gene expression, protein translation, and mRNA splicing. This project is jointly funded by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division in the Directorate of Biological Sciences and by the Established Program to Stimulate Competitive Research (EPSCoR).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.