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

描述（由申请人提供）：我们一直在使用出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）。我们发现，表达野生型a-syn （WT）或两种遗传突变体A53T或A30P中的任何一种的酵母细胞表现出几种凋亡标志物：磷脂酰丝氨酸的外化、活性氧的积累以及细胞色素c从线粒体释放到细胞质中。我们还发现，表达三种a-syn中的任何一种的酵母细胞对过氧化氢的杀伤非常敏感，过氧化氢是酵母中已知的细胞凋亡诱导剂。这一发现被用来创建一个独特的基因筛选。来自高拷贝文库的拯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文库-来自任何生物体的基因-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.