Transitions: Adopting a Protein Biochemistry Approach to Mechanistic Understanding of RNA Buffering

该项目将研究一种名为RNA缓冲的新调节机制，该机制可以保持信使RNA(MRNA)的稳定丰度，尽管它们的合成和衰变速度发生了变化。传统上，mRNA的丰度主要取决于合成(转录)的强度，衰变的影响很小，但动态RNA缓冲对基因表达的机制和影响尚不清楚。RNA缓冲是在酵母中发现的，直到最近才在多细胞生物体中被证明。这个项目将研究植物模型拟南芥中的RNA缓冲。研究这一现象很重要，因为它挑战了基因表达如何调控的传统观点，而且它的流行程度尚不清楚。由此产生的知识可能有助于推动生物技术的发展(例如，RNA缓冲可能会干扰操纵基因表达的策略，如设计作物抗性或生产药物)。这个项目还将为本科生提供培训机会，并为犹他大学和盐湖社区学院的学生提供关于RNA生物学的新教材，并为一个在线平台(learn.遗传学)制作新的教材。为了了解如何选择mRNAs进行RNA缓冲，以及细胞如何协调涉及细胞核和细胞质之间通讯的转录和衰退速率的变化，该项目将探索所涉及的蛋白质-蛋白质和蛋白质-RNA相互作用。将进行遗传和生化实验，以鉴定和表征与RNA缓冲相关的mRNA衰退成分相互作用的蛋白质(例如，静脉曲张蛋白质)，并将通过功能缺失突变研究来测试和验证候选蛋白质在RNA缓冲中的作用。除了研究成果，该项目还将为学习开发RNA生物学的教学材料。遗传学平台是学校和更广泛社区广泛使用的教育资源。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 project will investigate a novel regulatory mechanism called RNA buffering that maintains messenger RNAs (mRNA) at stable abundances despite changes in their rates of synthesis and decay. Traditionally, mRNA abundances are thought to depend primarily on the strength of synthesis (transcription) with a minor influence of decay, but the mechanism and impact of dynamic RNA buffering on gene expression is unclear. RNA buffering was discovered in yeast and has only recently been demonstrated in multicellular organisms. This project will investigate RNA buffering in the plant model Arabidopsis thaliana. Studying this phenomenon is important because it challenges conventional views of how gene expression is regulated, and because its prevalence is not known. The resulting knowledge could help advance biotechnology (e.g., RNA buffering might interfere with strategies to manipulate gene expression, such as engineering crop resistance or production of pharmaceuticals). This project will also offer training opportunities to undergraduates, and yield new educational materials on RNA biology for students at the University of Utah and Salt Lake Community college, and for an online platform (learn.genetics).To discover how mRNAs are selected for RNA buffering, and how cells coordinate changes in transcription and decay rates that involve communication between the nucleus and cytoplasm, this project will explore the protein-protein and protein-RNA interactions involved. Genetic and biochemical experiments will be performed to identify and characterize proteins that interact with mRNA decay components implicated in RNA buffering (e.g., VARICOSE protein), and candidates will be tested and validated for their roles in RNA buffering through loss-of-function mutational studies. In addition to research outcomes, the project will also develop instructional materials on RNA biology for the learn.genetics platform that is a widely used resource for education by schools and the broader community.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.