Competitive interplay of neuronal transcription factors via DNA methylation

神经元转录因子通过 DNA 甲基化的竞争性相互作用

基本信息

批准号：
9894858
负责人：
Junji Iwahara
金额：
$ 19.36万
依托单位：
UNIVERSITY OF TEXAS MED BR GALVESTON
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-03-15 至 2021-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9894858
关键词：
Binding Biochemical Biophysics Brain Brain Diseases CREB1 gene Calorimetry Cell Nucleus CpG Islands CpG dinucleotide Cyclic AMP Cyclic AMP-Responsive DNA-Binding Protein DNA DNA Methylation Drug Addiction Elements Epigenetic Process Fluorescence Spectroscopy Functional disorder Future Gene Activation Gene Expression Regulation Genes Genetic Predisposition to Disease Genomic DNA Goals Human Human Genome In Vitro Investigation Knowledge Learning Methyl-CpG-Binding Protein 2 Methylation Molecular Mus Mutation NMR Spectroscopy Neurons Pharmacology Phosphorylation Play Proteins Proteomics Research Research Project Grants Rett Syndrome Role Testing Therapeutic Titrations Transactivation Transcription Coactivator Transcription Repressor Transcriptional Activation Transcriptional Activation Domain Transcriptional Regulation autism spectrum disorder biophysical properties biophysical techniques demethylation epigenetic regulation gene repression genetic corepressor insight long term memory novel prevent promoter protein protein interaction screening therapeutic development transcription factor

项目摘要

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.

抽象的在大脑中，转录因子MECP2和CREB在活动依赖性基因中起着至关重要的作用长期记忆形成和学习的监管。这些蛋白质功能障碍与脑部疾病，例如RETT综合征和药物成瘾。 MECP2最初被确定为转录阻遏物专门识别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，但发现神经元MECP2激活在CPG岛启动子的控制下，DNA甲基化很少见的2,000多个基因。这该R21项目的总体目标是阐明MECP2充当转录的机制激活剂。该项目中检验的中心假设是MECP2通过新颖的双重激活许多基因通过与CREB直接相互作用以及通过间接相互作用的机制来防止CREB1 在非功能性诱饵中被隔离。该研究项目将追求以下特定的目的：1）在体外表征CREB和MECP2之间的直接相互作用，2）揭开间接这些蛋白质通过DNA甲基化之间的相互作用。研究小组将使用生物物理和生化方法检查双重机制。 MECP2关于反式激活的新知识将提供对大脑表观遗传调节的见解，并加深我们对脑疾病的理解。例如，我们对CREB-MECP2相互作用的生物物理特征将提供新的解释MECP2的RETT综合征突变的观点。通过此确定的方法项目有可能促进抑制CREB-MECP2的化合物的药理筛查未来药物成瘾治疗的相互作用。