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

该项目将研究一种称为RNA缓冲的新型调节机制，该机制使信使RNA（mRNA）保持稳定的丰度，尽管它们的合成和衰变速率发生变化。传统上，mRNA丰度被认为主要取决于合成（转录）的强度，衰变的影响较小，但动态RNA缓冲对基因表达的机制和影响尚不清楚。RNA缓冲在酵母中被发现，直到最近才在多细胞生物中得到证实。本计画将探讨植物模式拟南芥的RNA缓冲作用。研究这种现象很重要，因为它挑战了基因表达如何调节的传统观点，而且它的流行程度尚不清楚。由此产生的知识可以帮助推进生物技术（例如，RNA缓冲可能会干扰操纵基因表达的策略，例如工程作物抗性或药物生产。该项目还将为本科生提供培训机会，并为犹他州大学和盐湖社区学院的学生以及在线平台提供新的RNA生物学教育材料为了发现mRNA如何被选择用于RNA缓冲，以及细胞如何协调涉及细胞核和细胞质之间通信的转录和衰变速率的变化，这个项目将探索蛋白质-蛋白质和蛋白质-RNA的相互作用。将进行遗传和生物化学实验以鉴定和表征与RNA缓冲中涉及的mRNA衰变组分相互作用的蛋白质（例如，VARICOSE蛋白），并通过功能丧失突变研究测试和验证候选物在RNA缓冲中的作用。除了研究成果，该项目还将为learn.genetics平台开发RNA生物学教学材料，该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，并被认为值得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来支持。

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.