Elucidation of the molecular mechanims of Parkin-PINK1 triggered mitophagy by using budding yeast

在真核细胞中，异常线粒体通过一种称为线粒体自噬的过程被选择性地去除，而在哺乳动物细胞中，帕金森病的致病基因产物PINK1和Parkin介导了这一过程，而酵母细胞缺乏帕金森-PINK1系统。在这里，我们在酵母细胞中表达了Parkin和一种人工靶向线粒体外膜的PINK1融合蛋白。在胞浆中表达的Parkin被PINK1以依赖于PINK1激酶活性的方式募集到线粒体外膜。与哺乳动物细胞一样，被招募的帕金蛋白进一步磷酸化和泛素化。因此，人工表达Pakin-PINK1的酵母细胞可以为分析Pakin-PINK1系统的功能提供有用的工具。

In eukaryotic cells, aberrant mitochondria are selectively removed by a process called mitophagy, and PINK1 and Parkin, causative gene products for Parkinsonism, mediate this process in mammalian cells while yeast cells lack the Parkin-PINK1 system. Here we expressed Parkin and a PINK1 fusion protein that is artificially targeted to the mitochondrial outer membrane in yeast cells. Parkin expressed in the cytosol was recruited to the mitochondrial outer membrane by PINK1 in a manner that depends on the kinase activity of PINK1. The recruited Parkin was further phosphorylated and ubiquitinated like the case for mammalian cells. The yeast cells with artificially expressed Pakin-PINK1 can thus offer a useful tool to analyze the functions of the Parkin-PINK1 system.