A novel method to discover genes that protect cells from alpha synuclein

发现保护细胞免受α突触核蛋白侵害的基因的新方法

• 批准号: 7015120
• 负责人: Stephan N. Witt
• 金额: $ 16.31万
• 依托单位: LOUISIANA STATE UNIV HSC SHREVEPORT
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-01-10 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7015120
• 关键词: Parkinson' s disease; Saccharomyces cerevisiae; alpha synuclein; apoptosis; cytochrome c; cytoprotection; cytotoxicity; free radical oxygen; fungal genetics; genetic library; genetic screening; human genetic material tag; hydrogen peroxide; method development; phosphatidylserines

DESCRIPTION (provided by applicant): We have been using the budding yeast Saccharomyces cerevisiae to study the Parkinson's disease- related protein alpha synuclein (a-syn). We have discovered that yeast cells expressing wild-type a-syn (WT), or either of the two inherited mutants, A53T or A30P, exhibit several markers of apoptosis: externalization of phosphatidylserine, the accumulation of reactive oxygen species, and the release of cytochrome c from mitochondria into the cytosol. We also discovered that yeast cells expressing any of the three a-syns are very sensitive to killing by hydrogen peroxide, which is known inducer of apoptosis in yeast. This finding has been exploited to create a unique genetic screen. Genes from a high copy library that rescue hydrogen peroxide sensitivity may also suppress other traits of a-syn expression and provide insights into the mechanisms by which a-syn causes cell death. This method allows us to screen libraries of genes- genes from any organism-to discover genes that protect against a-syn toxicity.

描述（由申请人提供）：我们一直在使用酿酒酵母的萌芽酵母菌研究帕金森氏病相关的蛋白质αsnmuclein（A-Syn）。我们发现表达野生型A-Syn（WT）的酵母细胞，或两个遗传突变体A53T或A30P中的任何一个都表现出几种凋亡的标志：磷酸二甲酯的外部化，反应性氧的积累，反应性氧的积累，以及从线粒体中释放到CytoChrome C中。我们还发现，表达三种A-syn中任何一个的酵母细胞对被过氧化氢杀死非常敏感，这是酵母中凋亡的诱导剂。该发现已被利用以创建独特的遗传屏幕。从高复制文库中挽救过氧化氢敏感性的基因也可能抑制A-SYN表达的其他特征，并提供有关A-Syn导致细胞死亡的机制的见解。这种方法使我们能够从任何生物体中筛选基因基因的库，以发现可防止A-Syn毒性的基因。