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

描述(申请人提供)：我们一直在使用发芽酵母酿酒酵母研究帕金森病相关蛋白α突触核蛋白(a-syn)。我们已经发现，表达野生型a-syn(WT)的酵母细胞，或者两个遗传突变体A53T或A30P中的任何一个，都表现出几个凋亡的标志：磷脂酰丝氨酸外化，活性氧积累，以及细胞色素c从线粒体释放到胞浆中。我们还发现，表达这三种a-syn中任何一种的酵母细胞对过氧化氢非常敏感，过氧化氢是酵母细胞凋亡的诱导剂。这一发现被用来创造一种独特的基因筛查。来自高拷贝文库的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，并为a-syn导致细胞死亡的机制提供见解。这种方法使我们能够筛选基因库--来自任何生物体的基因库--以发现防止a-syn毒性的基因。

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.