Genomics/Proteomics of p53-mediated Neuronal Death

p53 介导的神经元死亡的基因组学/蛋白质组学

• 批准号: 6466242
• 负责人: MARK D JOHNSON
• 金额: $ 6.13万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-15 至 2003-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6466242
• 关键词: DNA damage; apoptosis; cell death; complementary DNA; fibroblasts; gene expression; gene targeting; genetically modified animals; genome; immunocytochemistry; in situ hybridization; laboratory mouse; mass spectrometry; messenger RNA; microarray technology; neural degeneration; neurons; neurotoxins; northern blottings; p53 gene /protein; protein structure function; proteomics; tissue /cell culture; transcription factor; western blottings

DESCRIPTION (provided by applicant): The p53 protein is a site-specific transactivator or repressor of transcription that promotes apoptosis, in part, by modulating the expression of select target genes. Studies have implicated p53 in the pathogenesis of neuronal cell death occurring after DNA damage orexcitotoxicity, or in neurodegenerative diseases such as Huntington?s Disease and Alzheimer?s Disease. However, the cellular consequences of p53 activation in neurons are poorly understood, and a comprehensive survey of changes in gene and protein expression in neurons after p53 activation is therefore needed. This project will combine the use of cDNA microarray technology and Fourier transform ion cyclotron resonance mass spectrometric analysis of the proteome to rapidly and comprehensively characterize p53-dependent changes in mRNA and protein expression in cortical neurons derived from p53+/+ and p53-/- mice after genotoxic or excitotoxic injury. Proteins essential to p53-mediated neuronal cell death will be identified and investigated further to elucidate their function. One such gene product, Peg3, is a Kruppel-type zinc finger protein and putative transcription factor that has recently been implicated in p53-mediated apoptosis in fibroblasts. Using transient transfection or exposure to DNA damaging agents, Peg3 expression will be manipulated in cortical neurons derived from wild type, p53-/- or bax-/- mice, and the effects on neuronal cell death will be assessed. The study of Peg3 will serve as a model for the investigation of other p53-regulated gene products in neurons. The candidate has earned both the M.D. and Ph.D. degrees from Harvard Medical School, and is now nearing completion of a residency in neurosurgery at the University of Washington, where he has recently been appointed as an Acting Instructor. The proposed research represents a continuation of the candidate?s longstanding interest in the physiology and survival of neurons. The skills acquired will assist the candidate in establishing an independent research career to study the determinants, of neuronal survival, differentiation and function.

描述（由申请人提供）：p53蛋白是一种位点特异性 促进凋亡的转录的反式激活子或阻遏子，部分地， 通过调节选择的靶基因的表达。研究表明， p53在DNA损伤或兴奋毒性后发生的神经元细胞死亡的发病机制中，或在神经退行性疾病如亨廷顿？老年痴呆症和阿尔茨海默病？的疾病。然而，p53在神经元中激活的细胞后果知之甚少，因此需要对p53激活后神经元中基因和蛋白表达的变化进行全面调查。本项目将联合收割机结合使用cDNA微阵列技术和傅立叶变换离子回旋共振质谱技术对蛋白质组进行分析 快速和全面地表征mRNA中的p53依赖性变化， 来自p53 +/+和p53-/-小鼠的皮质神经元中的蛋白表达 遗传毒性或兴奋性毒性损伤后。p53介导的蛋白质 神经元细胞死亡将被鉴定并进一步研究以阐明 它们的功能。其中一种基因产物Peg3是Kruppel型锌指 蛋白质和假定的转录因子，最近被牵连， p53介导的成纤维细胞凋亡。使用瞬时转染或暴露 对于DNA损伤剂，Peg3表达将在皮层神经元中被操纵， 来源于野生型、p53-/-或bax-/-小鼠，以及对神经元细胞的影响 死亡将被评估。对Peg3的研究将作为 神经元中其他p53调节基因产物的研究。候选 获得了医学博士学位和博士哈佛医学院的学位， 现在，他即将完成在芝加哥大学神经外科的住院实习。 华盛顿，他最近被任命为代理教官。的 申请的研究代表候选人的延续？长期以来的 对神经元的生理学和存活感兴趣。获得的技能将 协助候选人建立独立的研究生涯， 神经元存活、分化和功能的决定因素。