Coregulation of mRNA, tRNA, and rRNA species through RNA modifications

通过 RNA 修饰共同调节 mRNA、tRNA 和 rRNA 种类

• 批准号: 10190969
• 负责人: Fange Liu
• 金额: $ 40.01万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10190969
• 关键词: Affect; Binding Proteins; Biological Process; Catalysis; Cells; Code; Complex; Data; Deposition; Disease; Enzymes; Goals; In Vitro; Individual; Laboratories; Link; Mediating; Messenger RNA; Methylation; Methyltransferase; Modification; RNA; RNA Processing; Regulation; Research; Ribosomal RNA; Transfer RNA; Work; combinatorial; innovation; insight; preference; protein complex; recruit

Project Summary It has been demonstrated recently that a diverse set of enzyme-mediated modifications are found internally within RNAs which markedly influence the fate of RNAs in cells. Most of the RNA modifications are installed and removed by enzymes, termed writers (e.g., methyltransferases) and erasers (e.g., demethylases). Importantly, studies to date have focused solely on individual modifications on isolated RNA. However, recent data from my laboratory and others’ suggest cross-talk between modifications or the modifying enzymes influence both the depositions and consequences of several RNA modifications. Our preliminary data suggest that the level of one type of mRNA modification influences the level of the other, indicating that mRNA modifications may function in a previously undetermined combinatorial fashion. We also discovered that the level of a tRNA modification is concurrently regulated with a mRNA modification by a complex of RNA modifying enzymes. These data suggest the coordination between RNA modifying enzymes. However, the detailed mechanism of how these RNA modifying enzymes are coordinated and the biological function of the interplay of RNA modifications in tRNA and mRNA is not known. We discovered that a rRNA methyltransferase has dual enzymatic activities and install di-methylation on both rRNA and mRNA. Although the rRNA methyltransferase works equally well on rRNA and mRNA in vitro, the levels of mRNA modification are much lower in cells, suggesting that substrate preference is regulated. However, it is not known how this rRNA methyltransferase is controlled to achieve the substrate selectivity. To study whether a combinatorial modification code exists in mRNA, we will study the interplay between m3C and m6A in mRNA. Specifically, we will investigate whether the deposition of m3C affects m6A, and vice versa. We will also study the possible mechanism of the combinatorial mRNA modification in the recruitment of modification binding proteins, catalysis of RNA modifying enzymes, and the accessibility of the substrate. To understand the potential correlation between mRNA and tRNA through RNA modifications, we will investigate the interactions and the cellular consequence of the disturbed interactions of a protein complex of Trmt10A and YTHDF2. To understand how a dimethyladenosine methyltransferase regulates rRNA and mRNA modification, we will investigate the functions of dimethyladenosine in rRNA and mRNA, respectively. And then to study the regulatory mechanism by which this enzyme achieves substrate preference. Collectively, these innovative studies will provide fundamental insights into how multiple types of enzyme-mediated RNA modifications synergistically function.

项目概要 最近已经证明，在内部发现了多种酶介导的修饰 RNA 内的RNA 显着影响细胞中RNA 的命运。大多数 RNA 修饰已安装 并通过酶去除，称为写入酶（例如甲基转移酶）和擦除酶（例如去甲基酶）。 重要的是，迄今为止的研究仅集中于对分离 RNA 的个体修饰。然而，最近 我的实验室和其他实验室的数据表明修饰或修饰酶之间存在串扰 影响多种 RNA 修饰的沉积和后果。我们的初步数据表明 一种 mRNA 修饰的水平会影响另一种 mRNA 修饰的水平，表明 mRNA 修改可以以先前未确定的组合方式起作用。我们还发现， tRNA 修饰的水平与 RNA 复合物的 mRNA 修饰同时调节 修饰酶。这些数据表明 RNA 修饰酶之间存在协调作用。然而， 这些RNA修饰酶如何协调的详细机制以及这些酶的生物学功能 tRNA 和 mRNA 中 RNA 修饰的相互作用尚不清楚。我们发现一个rRNA 甲基转移酶具有双重酶活性，可在 rRNA 和 mRNA 上实现二甲基化。虽然 rRNA 甲基转移酶在体外对 rRNA 和 mRNA 的作用同样良好，mRNA 修饰的水平 细胞中的含量要低得多，表明底物偏好受到调节。然而，目前尚不清楚这是如何 控制rRNA甲基转移酶以实现底物选择性。研究组合是否 mRNA中存在修饰代码，我们将研究mRNA中m3C和m6A之间的相互作用。具体来说， 我们将研究 m3C 的沉积是否影响 m6A，反之亦然。我们也会研究可能的 组合 mRNA 修饰在修饰结合蛋白募集中的机制， RNA 修饰酶的催化作用以及底物的可及性。了解潜力 通过 RNA 修饰，mRNA 和 tRNA 之间的相关性，我们将研究相互作用和 Trmt10A 和 YTHDF2 蛋白质复合物相互作用受到干扰的细胞后果。到 了解二甲基腺苷甲基转移酶如何调节 rRNA 和 mRNA 修饰，我们将 分别研究二甲基腺苷在 rRNA 和 mRNA 中的功能。然后去研究 该酶实现底物偏好的调节机制。总的来说，这些创新 研究将为多种类型的酶介导的 RNA 修饰提供基本见解 发挥协同作用。