Regulatory Mechanisms of Arginine Methylation

摘要 精氨酸甲基化是最常见的翻译后修饰(PTM)之一，即 可与磷酸化和泛素化相媲美。蛋白质精氨酸甲基转移酶(PRMTs)对应于 产生三种甲基化精氨酸残基的“写入者”：甲基精氨酸(MMA)，不对称 二甲基精氨酸(ADMA)和对称性二甲基精氨酸(SDMA)。PRMT1是一种主要的I型酶 催化ADMA，而PRMT5是产生SDMA的主要II型酶。PRMT1/5 甲基化许多下游底物以调节各种基本的细胞过程，如 转录、DNA修复和细胞信号转导。经常观察到对PRMT1/5的放松管制 在多种癌症中，并与癌症患者的不良预后和生存相关。像许多其他人一样 精氨酸甲基化是一个可逆的过程。去甲基酶的作用是去除甲基的“橡皮”。 来自目标蛋白的组。此外，被称为“阅读器”的效应蛋白可与甲基精氨酸结合并进行调节。 细胞内的信号转导。尽管在过去的三十年里做出了巨大的努力，但有 精氨酸甲基化领域仍有许多悬而未决的问题/空白。PRMT活动是如何由 上游信号/监管机构？有没有特定的精氨酸脱甲基酶？许多人的读者是什么？ 精氨酸甲基化蛋白？在这项提案中，我们将探索三个项目来解决这些问题。 项目1将阐明氨基酸调节PRMT1亚细胞的分子机制。 定位、激活和功能，揭示了PRMT1的一个新的上游刺激/调节因子。项目2将 剖析泛素化途径在调节PRMT5中的作用，定义精氨酸之间的新的相互作用 甲基化和泛素化。项目3将确定新的脱甲基酶和精氨酸甲基化的读者， 填补精氨酸甲基化领域的关键空白。我们将使用一系列补充方法，包括 我们的生物化学、质谱分析、分子和细胞生物学以及小鼠模型 学习。我们的短期目标是通过完成 拟议的研究，长期目标是确定靶向精氨酸的新靶点/策略/抑制剂 癌症治疗中的甲基化信号通路。为了实现这些目标，我将致力于这个项目 51%的“研究成果”。

Abstract Arginine methylation is one of the most common posttranslational modifications (PTMs), which is comparable to phosphorylation and ubiquitination. Protein arginine methyltransferases (PRMTs) correspond to “writers” that generate three types of methylated arginine residues: monomethylarginine (MMA), asymmetric dimethylarginine (ADMA), and symmetric dimethylarginine (SDMA). PRMT1 is the main type I enzyme for catalyzing ADMA, while PRMT5 is the predominant type II enzyme for generating SDMA. PRMT1/5 methylates many downstream substrates to regulate a variety of fundamental cellular processes, such as transcription, DNA repair and cell signaling transduction. Deregulation of PRMT1/5 is frequently observed in various cancers and is correlated with poor prognosis and survival of cancer patients. Like many other PTMs, arginine methylation is a reversible process. Demethylases function as “erasers” to remove methyl groups from targeted proteins. In addition, effector proteins called “reader” bind to methylarginine and mediate signals transduction in cells. Although tremendous efforts have been made in the past three decades, there are still many outstanding questions/gaps in the field of arginine methylation. How is PRMT activity regulated by upstream signals/regulators? Are there specific arginine demethylases? What are the readers for numerous of arginine methylated proteins? In this proposal, we will explore three projects to address these questions. Project 1 will elucidate the molecular mechanism by which amino acids regulate PRMT1 subcellular localization, activation, and function, revealing a novel upstream stimulus/regulator of PRMT1. Project 2 will dissect roles of the ubiquitination pathway in regulation of PRMT5, defining a novel interplay between arginine methylation and ubiquitination. Project 3 will identify novel demethylases and readers of arginine methylation, filling the key gap in the field of arginine methylation. We will use a range of complementary methods including biochemistry, mass spectrometric (MS) analysis, molecular and cellular biology, and mouse models in our studies. Our short-term goal is to advance our understanding of arginine methylation biology by completing proposed studies, and log-term goal is to identify novel targets/strategies/inhibitors to target arginine methylation signaling pathway for cancer therapy. To achieve these goals, I will be committed to this program at 51% “research effort”.