Dynamic regulation of N6-methyladenosine sites in neuronal RNAs

描述（由申请人提供）：RNA的转录后修饰发生在所有真核生物中，包括脱氨基、假尿苷化和甲基化等过程。这种修饰对于RNA功能是非常宝贵的，因为它们可以影响从RNA剪接到正确翻译的过程。腺苷（N6-methyladenosine，或m6 A）N6位的甲基化是mRNA中最普遍的甲基修饰，尽管其丰富，但仍然知之甚少。虽然m6 A在大脑中以高水平存在，但其对神经元RNA功能的影响尚不清楚。为了研究m6 A在脑功能中的作用，本文提出的研究将使用RNA-seq识别神经元内易受活性依赖性调节的m6 A靶点。此外，该提案将研究调节神经元m6 A丰度的潜在甲基化和去甲基化途径。这些实验将是第一个在转录组水平上探索神经元内m6 A修饰的动态调节，并将为这种高度普遍的修饰如何促进神经元功能提供新的见解。此外，阐明影响m6 A丰度的细胞内途径将提供更好地理解神经元RNA响应神经元活性而调节的机制。通过确定m6 A在基本脑功能中的作用，我们可以推进我们对导致大脑疾病的分子和表观遗传事件的理解。 公共卫生相关性：在神经元中，RNA的修饰是一个动态过程，对大脑功能有重要影响。本文提出的研究将研究N6-甲基腺苷（m6 A），一种特别丰富的RNA修饰，在神经元功能中的作用。通过鉴定含有m6 A的神经元RNA和控制m6 A丰度的机制，我们可以获得重要的见解，了解这种高度流行的修饰如何有助于基本的脑功能，以及它的破坏如何导致神经系统疾病。

DESCRIPTION (provided by applicant): Post-transcriptional modification of RNA occurs in all eukaryotic organisms and includes processes such as deamination, pseudouridinylation, and methylation. Such modifications are invaluable for RNA function, as they can influence processes ranging from RNA splicing to proper translation. Methylation at the N6 position of adenosine (N6-methyladenosine, or m6A) is the most prevalent methyl modification in mRNA, yet despite its abundance, it remains poorly understood. Although m6A is present at high levels in the brain, its influence on the function of neuronal RNAs is unknown. To investigate the role of m6A in brain function, the research proposed here will identify m6A targets susceptible to activity-dependent regulation within neurons using RNA-seq. In addition, this proposal will investigate potential methylation and demethylation pathways that regulate neuronal m6A abundance. These experiments will be the first to explore the dynamic regulation of the m6A modification within neurons on a transcriptome-wide level and will provide novel insights into how this highly prevalent modification contributes to neuron function. Furthermore, elucidating the intracellular pathways that influence m6A abundance will provide a better understanding of the mechanisms through which neuronal RNAs are regulated in response to neuronal activity. By determining the role that m6A plays in basic brain function, we can propel our understanding of the molecular and epigenetic events that contribute to brain disorders. PUBLIC HEALTH RELEVANCE: In neurons, the modification of RNAs is a dynamic process which has important consequences for brain function. The research proposed here will investigate the role of N6-methyladenosine (m6A), a particularly abundant RNA modification, in neuronal function. By identifying the neuronal RNAs containing m6A and the mechanisms controlling m6A abundance, we can gain important insights into how this highly prevalent modification contributes to basic brain function and how its disruption might lead to neurological disorders.