Dynamic regulation of N6-methyladenosine sites in neuronal RNAs

描述(申请人提供)：RNA转录后修饰存在于所有真核生物中，包括脱氨基、假尿嘧啶基化和甲基化等过程。这些修饰对于RNA功能来说是无价的，因为它们可以影响从RNA剪接到正确翻译的各种过程。腺苷(N6-甲基腺苷，或m6A)的N6位甲基化是mRNA中最常见的甲基化修饰，尽管其含量丰富，但仍然知之甚少。虽然m6A在大脑中的水平很高，但它对神经元RNA功能的影响尚不清楚。为了研究m6A在脑功能中的作用，这里提出的研究将利用RNA-SEQ识别神经元内对活动依赖调节敏感的m6A靶点。此外，这项建议还将研究调节神经元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修饰的动态调节，并将为这一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丰度的细胞内途径将有助于更好地理解神经元RNA对神经元活动的调节机制。通过确定m6A在基本大脑功能中扮演的角色，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。 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。这里提出的研究将调查N6-甲基腺苷(M6A)，一种特别丰富的RNA修饰，在神经元功能中的作用。通过识别含有m6A的神经元RNA和控制m6A丰度的机制，我们可以获得重要的见解，了解这种高度普遍的修饰如何促进基本大脑功能，以及它的破坏可能如何导致神经疾病。

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.