m6A mRNA modifications and myogenesis

m6A mRNA 修饰和肌生成

基本信息

批准号：
10013127
负责人：
Nobuaki Kikyo
金额：
$ 16.94万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-09 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10013127
关键词：
3&apos Untranslated Regions 5&apos Untranslated Regions Affect Alternative Splicing Binding Proteins Biochemistry Biological Phenomena Biological Process Biology Cancer Biology Cell Differentiation process Cell Proliferation Cells Complex Consensus Sequence Data Set Family Family member Family suidae Foundations Future Genes Goals In Vitro Link Maps Messenger RNA Metabolism Methylation Methyltransferase Modification Molecular Muscle Muscle Cells Muscle Development Mutagenesis Myoblasts Myogenin Nuclear Export Nucleic Acid Binding Nucleotides Ontology Pattern Peptides Peptidylprolyl Isomerase Phenotype Phosphorylation Phosphorylation Site Play Pluripotent Stem Cells Population Proline Protein Family Proteins Publishing RNA Splicing RNA metabolism Reader Regulation Reporting Role Site Skeletal Muscle Solid Tacrolimus Binding Proteins Techniques Terminator Codon Translations Untranslated RNA base cancer cell cis trans isomerization genome-wide improved in vivo invention knock-down member muscle regeneration mutant myogenesis neurodevelopment novel recruit stem cell differentiation transcription factor

项目摘要

Project Summary N6-methyladenosine (m6A) is the most abundant internal modification of eukaryotic mRNA and long noncoding RNAs. m6A is induced by the Mettl3 complex most commonly near stop codons and 3’ UTRs in mRNAs. m6A recruits the YTH domain family of m6A-binding proteins, which then control almost every aspect of RNA metabolism, including alternative splicing, nuclear export, stability, and translation. Several thousand mRNA regions are modified by m6A in a given cell population, primarily at the consensus sequence of DRACH (D=A, G, or U; R=A or G; H=A, C, or U). Reflecting the diverse molecular functions of m6A, it also regulates a wide range of biological phenomena, such as cancer cell proliferation, neural development, and pluripotent stem cell differentiation. However, the roles of m6A in muscle cell differentiation remain largely elusive. The PI’s group recently found that one of the Fkbp family member of peptidyl prolyl isomerases interacts with the Mettl3 complex. In addition, depletion of the Fkbp inhibited myoblast differentiation and decreased the total level of m6A in the cells, both of which were recapitulated by Mettl3 depletion. At a molecular level, the Fkbp promotes cis-trans isomerization of Mettl3 and its related protein Mettl14 in vitro. Based on these findings, they hypothesized that the Fkbp regulates the m6A level of myoblast mRNAs through isomerization of Mettl3 and Mettl14. Three aims were proposed to investigate this hypothesis. Aim 1 will determine genome-wide distribution of m6A in myoblasts at a single nucleotide level and study how the pattern changes by depletion of the Fkbp. Aim 2 will characterize how m6A affects metabolism of several most abundantly modified myoblast-specific mRNAs. Aim 3 will study how Fkbp8 regulates Mettl3 and Mettl14, focusing on phosphorylation and isomerization of the proteins. These studies are expected to lay a solid foundation for a future study of the roles of m6A in muscle cells.

项目概要 N6-甲基腺苷 (m6A) 是真核 mRNA 和长非编码最丰富的内部修饰 RNA 是由 Mettl3 复合物诱导的，最常见的是 mRNA 中的终止密码子和 3’UTR。招募 m6A 结合蛋白的 YTH 结构域家族，然后控制 RNA 的几乎各个方面代谢，包括选择性剪接、核输出、稳定性和翻译。在给定的细胞群中，区域被 m6A 修饰，主要是在 DRACH 的共有序列处（D=A， G，或U；R=A或G；H=A，C，或U）。一系列生物现象，例如癌细胞增殖、神经发育和多能干细胞然而，m6A 在肌肉细胞分化中的作用在很大程度上仍然难以捉摸。最近发现肽基脯氨酰异构酶的 Fkbp 家族成员之一与 Mettl3 复合物相互作用。此外，Fkbp 的缺失抑制了成肌细胞分化并降低了 m6A 的总水平。细胞，这两种细胞均通过 Mettl3 耗竭进行概括。在分子水平上，Fkbp 促进顺反式。基于这些发现，他们发现了 Mettl3 及其相关蛋白 Mettl14 的体外异构化。 Fkbp 通过 Mettl3 和 Mettl14 的异构化调节成肌细胞 mRNA 的 m6A 水平三个目标。建议研究这一假设，目标 1 将确定成肌细胞中 m6A 的全基因组分布。在单核苷酸水平上进行分析，并研究 Fkbp 耗尽后模式如何变化。 Aim 3 将研究 m6A 如何影响几种修饰最丰富的成肌细胞特异性 mRNA 的代谢。 Fkbp8 如何调节 Mettl3 和 Mettl14，重点关注蛋白质的磷酸化和异构化。研究预计将为未来研究 m6A 在肌肉细胞中的作用奠定坚实的基础。