Elucidating the role of protein arginine methylation in regulating RNA-binding protein function

氨基酸精氨酸是在细胞中制造蛋白质的基石之一。在某些蛋白质中，精氨酸可以通过添加甲基进行化学修饰。然而，目前尚不清楚为什么会发生这种情况以及结果是什么。这个项目试图通过研究出芽酵母中的一种特定蛋白质来解决这个难题，这种蛋白质已经甲基化了精氨酸，并且似乎对允许细胞在不同环境中生长也很重要。关键目标是发现如何控制精氨酸甲基化过程以及精氨酸甲基化如何影响蛋白质功能。此外，该项目将通过在科研培训过程中整合教育外展，为提高科学教育水平和培养多层次的科学教育工作者提供平台。这包括针对K-12年级的具体外展活动。将强调征聘代表性不足的少数民族、妇女和残疾人。在真核生物中，许多rna结合蛋白携带蛋白精氨酸甲基化。这种翻译后修饰是由进化上保守的蛋白精氨酸甲基转移酶（PRMT）家族催化的。遗传学研究表明，蛋白质精氨酸甲基化有助于rna结合蛋白的功能，但尚不清楚这种翻译后修饰是如何调节的，以及这种调节的生物学后果是什么。该项目将通过研究出芽酵母酵母（Saccharomyces cerevisiae）中的一种模式rna结合蛋白（称为Npl3）来解决这些问题。在酵母中，Npl3在精氨酸上被主要的PRMT酶甲基化。当甲基化时，Npl3促进在生长周期中不同时间起作用的pre- mrna的剪接。该项目的目标是研究Npl3蛋白精氨酸甲基化调控的分子机制，以及这种调控在分子和细胞水平上的后果。使用分子生物学、生物化学、蛋白质组学和基因组学技术的多方面方法将被用于开展这些研究。这些结果有望提高我们对蛋白质精氨酸甲基化动力学及其对不同环境条件下细胞适应生长的影响的理解。本研究由生物科学理事会分子和细胞生物科学部的遗传机制和细胞动力学与功能项目共同资助。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The amino acid arginine is one of the building blocks for making proteins in the cell. In some proteins, arginines can be chemically modified by addition of methyl groups. However, it is not clear why this happens and what the outcomes are. This project seeks to solve this puzzle by studying a specific protein in budding yeast that has methylated arginines and that also appears to be important for allowing cells to grow in different environments. Key goals are to discover how the process of arginine methylation is controlled and how arginine methylation affects protein function. Also, the project will serve as a platform for improving science education and developing science educators across multiple levels, through integration of educational outreach during research training. This includes specific outreach activities aimed at K-12 level. Recruitment of underrepresented minority, women, and persons with disabilities will be emphasized. In eukaryotes, many RNA-binding proteins harbor protein arginine methylation. This post-translational modification is catalyzed by a family of evolutionarily conserved enzymes known as protein arginine methyltransferases (PRMT). Genetic studies have revealed that protein arginine methylation contributes to the function of RNA-binding proteins, but it is not clear how this post-translational modification is regulated, and what the biological consequences of such regulation are. This project will address these questions by studying a model RNA-binding protein (called Npl3) in the budding yeast Saccharomyces cerevisiae. Npl3 is methylated on arginines by the major PRMT enzyme in yeast. When methylated, Npl3 promotes splicing of pre-mRNAs that function at different times in the growth cycle. The goals of this project are to examine the molecular mechanisms by which protein arginine methylation in Npl3 is regulated and the consequences of such regulation at both molecular and cellular levels. A multi-faceted approach using molecular biology, biochemistry, proteomics, and genomic technologies will be utilized to carry out these studies. The outcomes are expected to improve our understanding of the dynamics of protein arginine methylation and its effects on cellular adaptation to growth under different environmental conditions.This research is co-funded by the Genetic Mechanisms and the Cellular Dynamics and Function programs in the Division of Molecular and Cellular Biosciences in the Directorate of Biological Sciences.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.