Genomics/Proteomics of p53-mediated Neuronal Death

描述(申请人提供)：P53蛋白是一种特定的位点 转录反式激活因子或转录抑制因子，部分促进细胞凋亡， 通过调节选定的靶基因的表达。研究表明， P53在DNA损伤或氧化毒性后神经细胞死亡的发病机制中，或在亨廷顿？S病和阿尔茨海默病？S病等神经退行性疾病中的作用。然而，对神经元中P53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质表达的变化进行全面的调查。本项目将结合使用cdna微阵列技术和傅立叶变换离子回旋共振质谱仪对蛋白质组进行分析。 为了快速和全面地表征P53依赖的mRNA和 P53+/+和P53-/-小鼠皮质神经元的蛋白表达 在遗传毒性或兴奋性毒性损伤之后。P53介导的蛋白质 神经细胞死亡将被确认并进一步研究以阐明 它们的功能。其中一种名为Peg3的基因产物是一种Kruppel类型的锌指 蛋白质和可能的转录因子，最近被认为与 P53介导的成纤维细胞凋亡。使用瞬时转染法或暴露法 对于DNA损伤剂，Peg3的表达将在皮质神经元中受到操纵 野生型P53-/-或Bax-/-小鼠来源及其对神经细胞的影响 将对死亡进行评估。对PEG3的研究将成为 神经元中其他受p53调控的基因产物的研究。候选人 已在哈佛医学院获得医学博士和博士学位， 现在即将完成在加州大学神经外科的实习 他最近在华盛顿被任命为代理教官。这个 拟议中的研究代表了候选人S长期任职的延续 对神经元的生理学和生存感兴趣。所获得的技能将 帮助应聘者建立独立的研究生涯进行研究 神经元存活、分化和功能的决定因素。

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.