Nuclear Role of the Proteasome in Synaptic Plasticity

蛋白酶体在突触可塑性中的核作用

• 批准号: 9171339
• 负责人: ASHOK N HEGDE
• 金额: $ 38.14万
• 依托单位: GEORGIA COLLEGE AND STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9171339
• 关键词: ATP phosphohydrolase; Affect; Antibodies; Applications Grants; Binding; Binding Sites; Bioinformatics; Biological; Biological Assay; Biological Models; Brain; Brain Diseases; Brain-Derived Neurotrophic Factor; Catalytic Domain; Cell Nucleus; Cells; Complex; Cyclic AMP-Responsive DNA-Binding Protein; DNA; DNA Sequence; Data; Disease; Dominant-Negative Mutation; Electrophysiology (science); Future; Gene Expression; Genes; Genetic Transcription; Genomic DNA; Goals; High-Throughput Nucleotide Sequencing; Hippocampus (Brain); Investigation; Knowledge; Learning; Link; Long-Term Potentiation; Maintenance; Mammalian Cell; Memory; Memory Loss; Messenger RNA; Methods; Molecular; Nervous System Physiology; Neuroglia; Neurons; Nuclear; Nuclear Localization Signal; Nuclear Translocation; Pathway interactions; Pattern; Phase; Physiological; Plasma Membrane Lipid Bilayer; Play; Polyubiquitin; Promoter Regions; Proteasome Binding; Proteins; Proteolysis; Recombinant Proteins; Reporting; Research; Role; Slice; Stimulus; Synapses; Synaptic plasticity; Techniques; Testing; Training; Transcription Coactivator; Transcriptional Regulation; Translations; Ubiquitin; Work; Yeasts; base; biological systems; biomedical scientist; cancer cell; cell type; chromatin immunoprecipitation; experience; gene function; innovation; long term memory; multicatalytic endopeptidase complex; next generation; next generation sequencing; novel; promoter; response; transcription factor; undergraduate student

Elucidating the mechanisms by which synapses are altered for long-term memory storage is crucial for understanding both normal and abnormal functions of the nervous system. Investigations over the years have established that new gene transcription and translation of the newly transcribed genes is required for maintenance of long-term synaptic plasticity and consolidation of long-term memory. Research during the last two decades has revealed that proteolysis by the ubiquitin-proteasome pathway (UPP) has an essential role in synaptic plasticity and memory. Much of the work on the UPP has been focused on its traditional function, namely, degradation of substrate proteins. It is now becoming clear that the proteasome has other roles in the cell such as regulation of transcription. Studies carried out on non-neuronal cell types have shown that the proteasome binds to promoters of actively transcribed genes and assists in transcription. A part of the proteasome called the 19S regulatory complex contains several ATPases among which Rpt1 has been shown to play a critical role in transcription. We will investigate the role of the proteasome in transcription by focusing on Rpt1. We will use hippocampal late phase long-term potentiation (L-LTP) as a model system for our studies. Our preliminary data show that Rpt1 translocates to the nucleus in hippocampal slices in response to L-LTP- inducing stimuli. Also, our results show that Rpt1 binds to specific promoters of the brain-derived neurotrophic factor (BDNF) gene. Our first aim is to use a high-throughput sequencing method in combination with chromatin immunoprecipitation with Rpt1 antibodies to identify the transcriptional targets of Rpt1. Our second aim is to test the hypothesis that the function of nuclear Rpt1 is critical for transcription and L-LTP maintenance. This project will lay the groundwork for elucidating the unconventional roles of the proteasome in transcription required for long-term synaptic plasticity which will have significant implications for understanding normal long-term memory as well as loss of memory seen in many diseases and disorders of the brain. Furthermore, by providing hands-on research experience to undergraduate students, this project will significantly enhance the training of future biomedical scientists.

阐明突触改变长期记忆存储的机制对于 了解神经系统的正常和异常功能。多年来的调查已 确定新基因的转录和新转录基因的翻译是 维持长期突触可塑性和巩固长期记忆。最近期间的研究 二十年来的研究表明，泛素蛋白酶体途径 (UPP) 的蛋白水解作用在 突触可塑性和记忆。 UPP 的大部分工作都集中在其传统功能上， 即底物蛋白的降解。现在越来越清楚的是，蛋白酶体在 细胞的转录调控等。对非神经元细胞类型进行的研究表明 蛋白酶体与活跃转录基因的启动子结合并协助转录。的一部分 称为 19S 调节复合物的蛋白酶体包含多种 ATP 酶，其中已显示 Rpt1 在转录中发挥关键作用。我们将通过关注蛋白酶体在转录中的作用来研究 在 Rpt1 上。我们将使用海马晚期长时程增强（L-LTP）作为我们研究的模型系统。 我们的初步数据表明，Rpt1 响应 L-LTP- 易位至海马切片中的细胞核。 诱发刺激。此外，我们的结果表明，Rpt1 与脑源性神经营养细胞的特定启动子结合。 因子（BDNF）基因。我们的首要目标是结合使用高通量测序方法 使用 Rpt1 抗体进行染色质免疫沉淀，以鉴定 Rpt1 的转录靶标。我们的第二个 目的是检验核 Rpt1 的功能对于转录和 L-LTP 至关重要的假设 维护。该项目将为阐明蛋白酶体在细胞中的非常规作用奠定基础。 长期突触可塑性所需的转录，这将对理解产生重大影响 正常的长期记忆以及许多大脑疾病和紊乱中出现的记忆丧失。 此外，通过为本科生提供实践研究经验，该项目将 显着加强未来生物医学科学家的培训。