Competitive interplay of neuronal transcription factors via DNA methylation

摘要 在大脑中，转录因子MeCP2和CREB在活性依赖基因中起着至关重要的作用 对长期记忆形成和学习的调节。这些蛋白质的功能障碍与 大脑疾病，如雷特综合征和药物成瘾。MeCP2最初被标识为一种 转录抑制因子，能识别DNA中甲基化的CpG二核苷酸。最近， 尽管缺乏任何已知的转录激活结构域，但已发现神经元MeCP2激活 超过2,000个基因在CpG岛启动子的控制下，DNA甲基化很少见。这个 这个R21项目的总体目标是阐明MeCP2作为转录因子的作用机制。 激活剂。这个项目检验的中心假设是MeCP2通过新的DUAL激活了许多基因 通过与CREB的直接相互作用和通过间接相互作用防止CREB1的机制 不会被隔离在不起作用的诱饵中。本研究项目将进行以下具体的研究 目的：1)研究CREB与MeCP2在体外的直接相互作用；2)揭示CREB与MeCP2的间接相互作用 这些蛋白质之间通过DNA甲基化相互作用。研究小组将使用生物物理和 生物化学方法来检查双重机制。关于MeCP2反式激活的新认识 将提供对大脑表观遗传调控的洞察，并加深我们对大脑疾病的理解。 例如，我们对CREB-MeCP2相互作用的生物物理表征将提供新的 MeCP2基因的Rett综合征突变的解释前景。通过这种方式建立的方法 该项目将有可能促进抑制CREB-MeCP2的化合物的药理筛选 未来药物成瘾治疗的互动。

Abstract In the brain, the transcription factors MeCP2 and CREB play crucial roles in activity-dependent gene regulation for long-term memory formation and learning. Dysfunction of these proteins is associated with brain disorders such as Rett syndrome and drug addiction. MeCP2 was originally identified as a transcriptional repressor that specifically recognizes methylated CpG dinucleotides in DNA. More recently, despite the lack of any known transcriptional activation domain, it was found that neuronal MeCP2 activates more than 2,000 genes under the control of CpG island promoters where DNA methylation is rare. The overall goal of this R21 project is to elucidate the mechanisms by which MeCP2 acts as a transcriptional activator. The central hypothesis tested in this project is that MeCP2 activates many genes by novel dual mechanisms through the direct interaction with CREB and through indirect interplay that prevents CREB1 from being sequestered in nonfunctional decoys. This research project will pursue the following specific aims: 1) characterize the direct interaction between CREB and MeCP2 in vitro and 2) unravel the indirect interplay between these proteins via DNA methylation. The research team will use biophysical and biochemical approaches to examine the dual mechanisms. New knowledge about transactivation by MeCP2 will provide insight into epigenetic regulation in the brain and deepen our understanding of brain disorders. For example, our biophysical characterizations of the CREB-MeCP2 interaction will provide new perspectives for interpreting Rett syndrome mutations of MeCP2. The approaches established through this project would potentially facilitate pharmacological screening of compounds that inhibit the CREB-MeCP2 interaction for drug addiction therapeutics in the future.