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.

阐明突触被改变以储存长期记忆的机制对于 了解神经系统的正常和异常功能。多年来的调查 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。研究在过去的 20年来的研究表明，通过泛素-蛋白酶体途径（UPP）的蛋白水解在 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，蛋白酶体在蛋白质合成中还有其他作用。 细胞，如转录调控。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19 S调节复合物的蛋白酶体含有几种ATP酶，其中Rpt 1已被证明是 在转录过程中起着至关重要的作用我们将研究蛋白酶体在转录中的作用， 在Rpt 1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，Rpt 1在海马脑片中响应L-LTP易位到核， 诱导刺激。此外，我们的研究结果表明，Rpt 1与脑源性神经营养因子的特异性启动子结合， 因子（BDNF）基因。我们的第一个目标是使用高通量测序方法， 染色质免疫沉淀与Rpt 1抗体，以确定Rpt 1的转录目标。我们的第二 目的是验证核Rpt 1的功能对转录和L-LTP至关重要的假设 上维护该项目将为阐明蛋白酶体在细胞内的非常规作用奠定基础。 转录所需的长期突触可塑性，这将有重大意义的理解 正常的长期记忆以及在许多疾病和大脑紊乱中看到的记忆丧失。 此外，通过为本科生提供实践研究经验，该项目将 大力加强对未来生物医学科学家的培训。