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、假尿嘧啶化和甲基化等过程。这些修饰对于RNA的功能是无价的，因为它们可以影响从RNA剪接到正确翻译的过程。腺苷N6位点的甲基化（N6-甲基腺苷，或m6A）是mRNA中最普遍的甲基修饰，尽管其丰富，但人们对其知之甚少。尽管m6A在大脑中存在高水平，但其对神经元rna功能的影响尚不清楚。为了研究m6A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动依赖性调节的m6A靶点。此外，本提案将研究调节神经元m6A丰度的潜在甲基化和去甲基化途径。这些实验将首次在转录组水平上探索神经元内m6A修饰的动态调控，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m6A丰度的细胞内通路将有助于更好地理解神经元rna在响应神经元活动时受到调节的机制。通过确定m6A在基本脑功能中的作用，我们可以推动我们对导致大脑疾病的分子和表观遗传事件的理解。

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.