Pum2-dependent translational regulation of a-SYN near mitochondria and contribution to the pathogenesis of Parkinson's disease

线粒体附近a-SYN的Pum2依赖性翻译调节及其对帕金森病发病机制的贡献

• 批准号: 10203457
• 负责人: YOON-SEONG KIM
• 金额: $ 34.13万
• 依托单位: RBHS-ROBERT WOOD JOHNSON MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10203457
• 关键词: 3' Untranslated Regions; Affect; Autopsy; Binding; Biochemical; Brain Diseases; CRISPR/Cas technology; Complex; DNA Sequence Alteration; Data Reporting; Development; Elements; FYN gene; Family; Goals; Human; Image; In Vitro; Inner mitochondrial membrane; Label; Length; Link; Mediating; Messenger RNA; Mission; Mitochondria; Mitochondrial Proteins; Molecular; Monitor; Morphology; NADH dehydrogenase (ubiquinone); National Institute of Neurological Disorders and Stroke; Nuclear; Outer Mitochondrial Membrane; Oxidative Stress; Parkinson Disease; Pathogenesis; Play; Polyribosomes; Protein translocation; Proteins; Public Health; Quality of life; RNA Binding; RNA Recognition Motif; RNA-Binding Proteins; Reactive Oxygen Species; Regulation; Regulatory Element; Research; Role; Single Nucleotide Polymorphism; Surface; Techniques; Testing; Toxic effect; Trans-Activators; Transcript; Translating; Translational Regulation; Translational Repression; Translations; United States National Institutes of Health; Untranslated Regions; Yeasts; alpha synuclein; base; genome editing; improved; in vivo; mitochondrial dysfunction; novel; novel therapeutics; overexpression; paralogous gene

Alpha-synuclein (α-SYN) and mitochondrial dysfunction are two central components in Parkinson's disease (PD) pathogenesis. Mitochondrial dysfunction is a common feature of the many iterations of PD pathogenesis and α-SYN toxicity seems to affect mitochondria most significantly. Complex interplay between α-SYN and mitochondria has been widely observed. While the intricate crosstalk between mitochondria and α-SYN is poorly understood, our preliminary studies suggest that the 3'-untranslated region (3'-UTR) of α-SYN mRNA plays a key role in translational regulation of α-SYN near mitochondria. Our preliminary findings demonstrate that 1) α-SYN mRNA is localized to the mitochondrial surface where its translation is initiated by mitochondrial ROS; 2) this translational control is governed by Pum2, a RNA-binding translational repressor, which binds to the 3'-untranslated region (3'-UTR) of α-SYN transcript; 3) interestingly, mitochondrial Pum2 levels in post-mortem PD brain were significantly lower compared to control subjects, while α-SYN levels were opposite, implying Pum2’s repressive role on α-SYN near mitochondria. In addition, recent studies showing the association of single nucleotide polymorphisms in the α-SYN 3'-UTR with PD strongly suggest that 3`-UTR-mediated regulation of α-SYN could become a critical player in PD pathogenesis. Our central hypothesis is that deregulation of Pum2-mediated α-SYN translational repression on the outer surface of mitochondria contributes to mitochondrial dysfunction observed in PD. The following three specific aims will be pursued: In Aim 1, both the cis-regulatory elements and the trans- factors responsible for mitochondrial localization of α-SYN will be identified. In Aim 2, it will be determined how mitochondrial ROS controls Pum2-mediated translation of α-SYN mRNA and the roles of newly synthesized α- SYN. In Aim 3, it will be investigated whether PD-associated SNPs in the 3'-UTR of α-SYN cause changes in Pum2 binding, translocation of the protein to mitochondria, and mitochondrial functions. The successful completion of this project could create a paradigm shift in our understanding of molecular mechanisms that control α-SYN expression near mitochondria in PD pathogenesis by elucidating the role of Pum2 and the 3'-UTR of α-SYN in translational regulation

α-突触核蛋白（α-SYN）和线粒体功能障碍是帕金森病（PD）发病机制的两个核心组成部分。线粒体功能障碍是PD发病机制多次迭代的共同特征，α-SYN毒性似乎对线粒体的影响最显著。已广泛观察到α-SYN和线粒体之间的复杂相互作用。虽然线粒体和α-SYN之间复杂的串扰知之甚少，但我们的初步研究表明，α-SYN mRNA的3 '-非翻译区（3'-UTR）在线粒体附近的α-SYN翻译调控中起着关键作用。我们的初步研究结果表明：（1）α-SYN mRNA定位于线粒体表面，其翻译由线粒体ROS启动，（2）这种翻译控制由Pum 2控制，Pum 2是一种RNA结合的翻译抑制因子，它与α-SYN转录本的3 '-非翻译区（3'-UTR）结合; 3）PD脑组织线粒体Pum 2水平显著低于对照组，而α-SYN水平则相反，提示Pum 2对线粒体附近的α-SYN具有抑制作用。此外，最近的研究显示α-SYN 3 '-UTR中的单核苷酸多态性与PD的相关性，强烈表明3'-UTR介导的α-SYN调节可能成为PD发病机制中的关键参与者。我们的中心假设是Pum 2介导的线粒体外表面α-SYN翻译抑制的失调导致PD中观察到的线粒体功能障碍。将追求以下三个具体目标：在目标1中，将鉴定负责α-SYN线粒体定位的顺式调节元件和反式因子。目的2将确定线粒体ROS如何控制Pum 2介导的α-SYN mRNA翻译以及新合成的α- SYN的作用。在目的3中，将研究α-SYN的3 '-UTR中的PD相关SNP是否引起Pum 2结合、蛋白质向线粒体的易位和线粒体功能的变化。该项目的成功完成可能会通过阐明Pum 2和α-SYN的3 '-UTR在翻译调控中的作用，为我们理解PD发病机制中控制线粒体附近α-SYN表达的分子机制带来范式转变