Recognition and mechanism of N6-methyl adenosine modifications

N6-甲基腺苷修饰的识别和机制

• 批准号: 10426289
• 负责人: TAO PAN
• 金额: $ 32万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10426289
• 关键词: Address; Adenosine; Affect; Alternative Splicing; Apoptosis; Binding; Biochemical; Biological Process; C-terminal; Cells; Complex; Development; Enzymes; Eukaryota; Exclusion; Exons; Family; Gene Expression; Genetic Transcription; Human; Immune response; Investigation; Knock-out; Laboratories; Mammals; Messenger RNA; Methyltransferase; Modeling; Modification; Molecular; Nuclear; Nuclear Export; Pattern; Proteins; RNA; RNA Binding; RNA Polymerase II; RNA Splicing; Reader; Reading; Regulation; Research; Research Personnel; Ribonucleoproteins; Role; Site; Structure; Testing; Translational Regulation; Translations; Untranslated RNA; Work; cell type; epitranscriptomics; frontier; knock-down; mRNA Precursor; mRNA Stability; mRNA Transcript Degradation; molecular modeling; transcriptome

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.

项目摘要/摘要 N6-甲基 - 腺苷（M6A）是信使和长期非编码中最丰富的内部修饰 RNA。这种修饰发生在mRNA中的许多位点，其功能包括剪接，导出，定位， 稳定性，翻译和免疫反应。 M6A修饰的功能咨询已被强烈 自2011年以来进行了研究，并已成为新表演组学领域不可或缺的组成部分。研究 来自我们的实验室和其他实验室，表明M6A修改通过与 特定的细胞蛋白称为M6A读取器。该建议调查了M6A读取器的生物学功能 蛋白质并解决潜在的分子和细胞机制。我们提出的研究将 通过两个核定位的M6A读取器蛋白及其在细胞中建立M6A功能的分子模型 作用机理。 AIM 1将研究共转录，M6A依赖性调节的分子和细胞机制 mRNA替代剪接。我们将通过RNA在HNRNPG/M6A依赖性控制上测试我们的模型 聚合酶II暂停和使用靶向方法的新生RNA结构的影响。 AIM 2将通过低复杂区域和 组装与M6A修饰的RNA和替代剪接相互作用的影响。我们将测试分子 使用生化和细胞方法在这些方面进行模型。 AIM 3将研究两个调节的两个核位置M6A读取器蛋白的M6A依赖机制 整个转录组的替代剪接。我们将阐明这两种蛋白质的相互作用 外显子目标以及如何发生。