Recognition and mechanism of N6-methyl adenosine modifications

N6-甲基腺苷修饰的识别和机制

• 批准号: 10627015
• 负责人: TAO PAN
• 金额: $ 8.28万
• 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/10627015
• 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.

项目概要/摘要 N6-甲基-腺苷（m6A）是信使和长非编码中最丰富的内部修饰 核糖核酸。这种修饰发生在 mRNA 的许多位点，其功能包括剪接、输出、定位、 稳定性、翻译和免疫反应。 m6A 修饰的功能研究已经深入开展 自 2011 年开始研究，现已成为表观转录组学新领域不可或缺的组成部分。研究 我们的实验室和其他人的研究表明，m6A 修饰通过与 称为 m6A 阅读器的特定细胞蛋白。该提案研究了 m6A reader 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 阅读器蛋白及其它们建立细胞中 m6A 功能的分子模型 行动机制。 目标 1 将研究共转录、m6A 依赖性调节的分子和细胞机制 mRNA 选择性剪接。我们将通过 RNA 测试 HNRNPG/m6A 依赖性控制的模型 使用靶向方法聚合酶 II 暂停和新生 RNA 结构的影响。 目标2将通过低复杂性区域和低复杂度区域研究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。我们将测试分子 使用生化和细胞方法对这些方面进行建模。 目标 3 将研究两种核定位 m6A 阅读器蛋白的 m6A 依赖性机制，这些蛋白调节 全转录组选择性剪接。我们将阐明这两种蛋白质之间的相互作用 外显子目标以及这是如何发生的。

项目成果

Quantitative probing of glycosylated queuosine modifications in tRNA.

tRNA 中糖基化 queuosine 修饰的定量探测。

• DOI: 10.1016/bs.mie.2021.06.003
• 发表时间: 2021
• 期刊: Methods in enzymology
• 作者: Zhang,Wen;Pan,Tao
• 通讯作者: Pan,Tao

Single-read tRNA-seq analysis reveals coordination of tRNA modification and aminoacylation and fragmentation.

单读 tRNA-seq 分析揭示了 tRNA 修饰与氨酰化和片段化的协调。

• DOI: 10.1093/nar/gkac1185
• 发表时间: 2023-02-22
• 期刊: NUCLEIC ACIDS RESEARCH
• 影响因子: 14.9
• 作者: Hernandez-Alias, Xavier;Katanski, Christopher D.;Zhang, Wen;Assari, Mahdi;Watkins, Christopher P.;Schaefer, Martin H.;Serrano, Luis;Pan, Tao
• 通讯作者: Pan, Tao

A high-throughput screening method for evolving a demethylase enzyme with improved and new functionalities.

• DOI: 10.1093/nar/gkaa1213
• 发表时间: 2021-03-18
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Wang Y;Katanski CD;Watkins C;Pan JN;Dai Q;Jiang Z;Pan T
• 通讯作者: Pan T

Detection and quantification of glycosylated queuosine modified tRNAs by acid denaturing and APB gels.

通过酸变性和 APB 凝胶检测和定量糖基化 queuosine 修饰的 tRNA。

• DOI: 10.1261/rna.075556.120
• 发表时间: 2020
• 期刊: RNA (New York, N.Y.)
• 作者: Zhang,Wen;Xu,Ruyi;Matuszek,Żaneta;Cai,Zhen;Pan,Tao
• 通讯作者: Pan,Tao

N6-methyladenosine alters RNA structure to regulate binding of a low-complexity protein.

• DOI: 10.1093/nar/gkx141
• 发表时间: 2017-06-02
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Liu N;Zhou KI;Parisien M;Dai Q;Diatchenko L;Pan T
• 通讯作者: Pan T