N6-Methyladenosine RNA Methylation in Myogenesis

项目总结/摘要 在分子水平上的关键肌源性过程还不完全清楚，阻碍了肌源性疾病的发展。 许多肌肉退行性疾病的治疗干预。为了填补这一知识空白，我们 研究一种新兴的转录后基因调控模式，称为表转录组学， 在骨骼肌发生（成肌细胞分化和肌肉再生）中的作用。表位转录组学是一种动态的 通过对mRNA进行可逆的化学修饰来调节mRNA稳定性、剪接和翻译的过程。 N6-甲基腺苷（m6 A）是最丰富的表位转录组标记。m6 A表观标记是 由甲基转移酶样3（胃L3）和甲基转移酶样14（胃L14）安装，并由 烷基化修复同源物5（ALKBH 5）和脂肪量和肥胖相关（FTO）。一组阅读蛋白 选择性识别m6 A修饰的mRNA并控制它们的命运。我们最近的研究表明，m6 A mRNA 甲基化调节成肌细胞分化和骨骼肌再生， 在这些过程中，epitranscriptomic writers和erasers受到严格的控制。我们还观察到 在转录组水平上的m6 A表型标记。这些研究共同表明，m6 A mRNA甲基化通过调节关键的分子途径在正常的肌生成过程中起着核心作用。 在这里，我们建议识别参与这一过程的分子参与者，从而获得更深入的机制 见解.我们将进行一项创新的全转录组分析，以确定m6 A的直接mRNA靶点。 甲基化在生肌过程中的作用。此外，我们将机械地描述一个选择的子集， 优先考虑m6 A靶mRNA，这两个先前与肌生成相关的，以及新的，以检查它们的 参与肌生成。我们提出的研究将通过阐明一个基本的 m6 A mRNA甲基化在关键肌形成过程中的分子机制。

PROJECT SUMMARY/ABSTRACT The key myogenic processes at the molecular level are incompletely understood, hindering the development of therapeutic interventions for numerous muscle degenerative diseases. To fill this knowledge gap, we have been investigating how an emerging mode of post-transcriptional gene regulation known as epitranscriptomics, plays a role in skeletal myogenesis (myoblast differentiation and muscle regeneration). Epitranscriptomics is a dynamic process that regulates mRNA stability, splicing, and translation by reversible chemical modifications on mRNAs. N6-methyladenosine (m6A) is the most abundant epitranscriptomic mark. The m6A epitranscriptomic mark is installed by methyltransferase like 3 (METTL3) and methyltransferase like 14 (METTL14), and erased by alkylation repair homolog 5 (ALKBH5) and fat mass- and obesity-associated (FTO). A group of reader proteins selectively recognize m6A-modified mRNAs and control their fates. Our recent studies show that m6A mRNA methylation regulates myoblast differentiation and skeletal muscle regeneration and that the levels of epitranscriptomic writers and erasers are tightly regulated during these processes. We also observed changes in m6A epitranscriptomic marks at the transcriptome-wide level. These studies collectively suggest that m6A mRNA methylation plays a central role in normal myogenic processes by regulating critical molecular pathways. Here, we propose to identify molecular players involved in this process and thus gain deeper mechanistic insights. We will perform an innovative transcriptome-wide analysis to identify direct mRNA targets of m6A methylation in myogenic processes. Additionally, we will mechanistically characterize a select subset of prioritized m6A target mRNAs, both previously associated with myogenesis, as well as novel, to examine their involvement in myogenesis. Our proposed research will make a significant impact by elucidating a fundamental molecular mechanism of m6A mRNA methylation in the key myogenic processes.