Pum2-dependent translational regulation of a-SYN near mitochondria in neurites

神经突线粒体附近 a-SYN 的 Pum2 依赖性翻译调节

• 批准号: 8772853
• 负责人: YOON-SEONG KIM
• 金额: $ 18.1万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8772853
• 关键词: 3' Untranslated Regions; Affect; Axon; Binding; Cells; Complex; Consensus; Consensus Sequence; Cytoplasmic Protein; Dendrites; Development; Disease; Dose; Family; Genes; Growth; Hereditary Disease; Homologous Gene; Homologous Protein; Human; In Vitro; Investigation; Lewy Bodies; Luciferases; Mediating; Messenger RNA; Mitochondria; Molecular; Morphogenesis; Mutation; NADH dehydrogenase (ubiquinone); Neurites; Neurons; Orthologous Gene; Oxidative Stress; Parkinson Disease; Pathogenesis; Physiological; Play; Presynaptic Terminals; Process; Protein Binding; Protein Biosynthesis; Proteins; Proteome; RNA Recognition Motif; RNA-Binding Proteins; Reactive Oxygen Species; Regulation; Reporter; Role; Rotenone; Sequence Analysis; Sequence Deletion; Signal Transduction; Staging; Synapses; System; Techniques; Translational Regulation; Translational Repression; Translations; Untranslated Regions; Yeasts; alpha synuclein; base; dopaminergic neuron; in vivo; inhibitor/antagonist; insight; interest; knock-down; mitochondrial dysfunction; mitochondrial messenger RNA; novel; novel therapeutics; nucleocytoplasmic transport; overexpression; protein aggregate; public health relevance; response

DESCRIPTION (provided by applicant): The presence of alpha-synuclein (alpha-SYN) aggregate in Lewy neurites (LNs) and Lewy bodies (LBs), the pathological hallmarks of Parkinson's disease (PD), and mitochondrial dysfunction are two central components in PD pathogenesis. Known physiological roles of alpha-SYN are limited in synaptic terminals, raising the possibility that alpha-SYN mRNA is transported to neurites and its translation is locally controlled. In fact, we have recently found that alpha-SYN mRNA is translocated into neurites of fully differentiated human dopaminergic neuronal cells. We also identified that Pum2, a paralogue of mammalian PUF family RNA binding protein, binds to a PUM binding motif of the alpha-SYN 3'UTR and is responsible for neuritic localization as well as translational repression of alpha-SYN mRNA. Mitochondrial ROS generated by respiratory complex I inhibition led to local translation of alpha-SYN from mRNA associated with mitochondria, suggesting that Pum2 might transmit mitochondrial signal to alpha-SYN translation. Although it is obvious that there is an inter- relationship between mitochondria and alpha-SYN underlying the development of disease, molecular mechanisms governing this cross-talk remain elusive. Our central hypothesis is that Pum2 is responsible for alpha-SYN mRNA transport to the juxta-mitochondrial compartment, alpha-SYN translational regulation and aggregation in neurites. The objectives of this project are to understand the interplay between alpha-SYN and mitochondria in neurites through Pum2-mediated translational regulation of mitochondria-associated alpha-SYN mRNA. To achieve these objectives we propose the following three specific aims: Aim 1. Examine the role of Pum2 in mitochondrial localization of alpha-SYN mRNA. Aim 2. Examine how mitochondrial ROS control alpha-SYN translation. Aim 3. Determine how newly synthesized alpha-SYN near mitochondria affects mitochondrial fragmentation and alpha-SYN aggregation in neurites. The successful completion of the project will bring a paradigm shift in our understanding of molecular mechanisms that control alpha-SYN level in neurites and how mitochondrial ROS contribute to this alpha-SYN regulation. The results expected from the project will open new avenues to understand 1) 3'UTR-dependent posttranscriptional regulation of alpha-SYN in neurites and Pum2's role in this process, 2) intimate crosstalk between alpha-SYN and mitochondria in neurites and 3) its contribution to mitochondrial fragmentation and LN formation.

描述（由申请人提供）：路易神经突（LN）和路易体（LB）中存在α-突触核蛋白（α-SYN）聚集体、帕金森病（PD）的病理学标志和线粒体功能障碍是PD发病机制的两个核心组成部分。已知的alpha-SYN的生理作用在突触末梢中是有限的，这增加了alpha-SYN mRNA被转运到神经突并且其翻译被局部控制的可能性。事实上，我们最近发现，α-SYN mRNA易位到完全分化的人多巴胺能神经元细胞的神经突中。我们还鉴定了哺乳动物PUF家族RNA结合蛋白Pum 2与α-SYN 3 'UTR的PUM结合基序结合，并负责神经炎定位以及α-SYN mRNA的翻译抑制。呼吸复合物I抑制产生的线粒体ROS导致与线粒体相关的mRNA的α-SYN的局部翻译，这表明Pum 2可能将线粒体信号传递到α-SYN翻译。虽然线粒体和α-SYN之间存在疾病发展的内在关系是显而易见的，但控制这种串扰的分子机制仍然难以捉摸。 我们的中心假设是Pum 2负责alpha-SYN mRNA转运到线粒体旁区室、alpha-SYN翻译调节和神经突聚集。本项目的目的是通过Pum 2介导的对神经元相关的α-SYN mRNA的翻译调节来了解α-SYN和神经突中线粒体之间的相互作用。为了实现这些目标，我们提出了以下三个具体目标：目标1。检查Pum 2在alpha-SYN mRNA的线粒体定位中的作用。目标二。研究线粒体ROS如何控制alpha-SYN翻译。目标3.确定线粒体附近新合成的α-SYN如何影响线粒体片段化和神经突中的α-SYN聚集。 该项目的成功完成将使我们对控制神经突中α-SYN水平的分子机制以及线粒体ROS如何促进这种α-SYN调节的理解发生范式转变。该项目的预期结果将开辟新的途径，以了解1）神经突中α-SYN的3 'UTR依赖性转录后调控和Pum 2在此过程中的作用，2）神经突中α-SYN和线粒体之间的密切串扰，以及3）其对线粒体片段化和LN形成的贡献。