m6A mRNA modifications and myogenesis

m6A mRNA 修饰和肌生成

• 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
• 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' Untranslated Regions; 5' 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

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.

项目摘要 N6-甲基腺苷（M6A）是真核mRNA的最丰富的内部修饰和长期不编码 RNA。 M6A是由METTL3复合物诱导的，最常见的是在MRNA中靠近停止密码子和3'UTR。 M6A 招募M6A结合蛋白的Yth领域家族，然后控制RNA的几乎所有方面 代谢，包括替代剪接，核输出，稳定性和翻译。几千个mRNA 区域在给定的细胞群中通过M6A修饰，主要是在Drach的共有序列（d = a， G，或U; r = a或g; h = a，c或u）。反映M6A的潜水员分子功能，它也调节 一系列生物学现象，例如癌细胞增殖，神经元发育和多能干细胞 分化。但是，M6a在肌肉细胞分化中的作用在很大程度上仍然是弹性。 Pi的小组 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的耗竭抑制了成肌细胞的分化，并降低了M6A的总水平 细胞，这两种细胞都是通过Mettl3耗竭概括的。在分子水平上，FKBP促进顺式传播 MetTL3及其相关蛋白METTL14体外的异构化。基于这些发现，他们假设 FKBP通过METTL3和METTL14的异构化来调节成肌细胞mRNA的M6a水平。三个目标 我们被提议研究这一假设。 AIM 1将确定肌细胞中M6A的全基因组分布 在单个核苷酸水平上，研究模式如何随FKBP的耗竭而变化。 AIM 2将表征 M6A如何影响几种最丰富的肌细胞特异性mRNA的代谢。 AIM 3将学习 FKBP8如何调节METTL3和METTL14，侧重于蛋白质的磷酸化和异构化。这些 预计研究将为未来对M6A在肌肉细胞中的作用的研究奠定坚实的基础。