Recognition and mechanism of N6-methyl adenosine modifications

项目摘要/摘要 N6-甲基腺苷(M6A)是信使和长非编码中含量最丰富的内部修饰 核糖核酸。这种修饰存在于mRNA中的许多位点，其功能包括剪接、输出、定位、 稳定性、转译和免疫反应。关于M6A修饰的功能研究一直很激烈 自2011年以来进行了研究，并已成为新的表位转录组学领域的组成部分。研究 来自我们实验室和其他人的研究表明，m6A修饰通过与以下物质的相互作用发挥作用 称为m6A阅读器的特定细胞蛋白。该方案研究了M6A阅读器的生物学功能 并阐述了潜在的分子和细胞机制。我们提议的研究将 通过两种核定位的m6A阅读器蛋白及其相互作用建立细胞内m6A功能的分子模型 行动机制。 目的1将研究共转录的分子和细胞机制，m6A依赖的调节。 信使核糖核酸的选择性剪接。我们将通过RNA测试我们的模型对HNRNPG/m6A依赖的控制 聚合酶II的暂停和使用靶向方法对新生RNA结构的影响。 目的2将通过低复杂性区域研究HNRNPG组装的功能和机制 组装对与m6A修饰的RNA相互作用和选择性剪接的影响。我们将测试一种分子 使用生物化学和细胞方法对这些方面进行建模。 目的3将研究两个核定位的m6A阅读器蛋白调节m6A的依赖机制 转录组范围的选择性剪接。我们将在特定的情况下阐明这两种蛋白质之间的相互作用 外显子靶标及其如何发生。

Project Summary/Abstract N6-methyl-adenosine (m6A) is the most abundant internal modifications in messenger and long non-coding RNA. This modification occurs in many sites in mRNA with functions including splicing, export, localization, stability, translation, and immune response. Functional inquiries of the m6A modification have been intensely studied since 2011 and have become an integral component of the new field of epitranscriptomics. Studies from our laboratories and others indicate that m6A modifications exert their function through interactions with specific cellular proteins termed m6A readers. This proposal investigates the biological function of m6A reader proteins and addresses the underlying molecular and cellular mechanisms. Our proposed research will establish molecular models of m6A function in cells through two nuclear-localized m6A reader proteins and their mechanisms of action. Aim 1 will investigate the molecular and cellular mechanisms of co-transcriptional, m6A-dependent regulation of mRNA alternative splicing. We will test our model on HNRNPG/m6A-dependent control through RNA polymerase II pausing and the effect of nascent RNA structure using targeted approaches. Aim 2 will study the function and mechanism of HNRNPG assembly through a low-complexity region and the effect of assembly on interacting with m6A-modified RNA and alternative splicing. We will test a molecular model on these aspects using biochemical and cellular approaches. Aim 3 will study the m6A-dependent mechanism of two nuclear-localized m6A reader proteins that regulate transcriptome-wide alternative splicing. We will elucidate the interplay among these two proteins on specific exon targets and how this occurs.