Department of Biological Sciences, The University of Alabama, Tuscaloosa, AL 35487, United States.
Biochem Biophys Res Commun. 2010 Aug 20;399(2):123-8. doi: 10.1016/j.bbrc.2010.07.014. Epub 2010 Jul 23.
Rotenone is a widely used pesticide that induces Parkinson's disease-like symptoms in rats and death of dopaminergic neurons in culture. Although rotenone is a potent inhibitor of complex I of the mitochondrial electron transport chain, it can induce death of dopaminergic neurons independently of complex I inhibition. Here we describe effects of rotenone in the fission yeast, Schizosaccharomyces pombe, which lacks complex I and carries out rotenone-insensitive cellular respiration. We show that rotenone induces generation of reactive oxygen species (ROS) as well as fragmentation of mitochondrial networks in treated S. pombe cells. While rotenone is only modestly inhibitory to growth of wild type S. pombe cells, it is strongly inhibitory to growth of mutants lacking the ERK-type MAP kinase, Pmk1, or protein kinase A (PKA). In contrast, cells lacking the p38 MAP kinase, Spc1, exhibit modest resistance to rotenone. Consistent with these findings, we provide evidence that Pmk1 and PKA, but not Spc1, are required for clearance of ROS in rotenone treated S. pombe cells. Our results demonstrate the usefulness of S. pombe for elucidating complex I-independent molecular targets of rotenone as well as mechanisms conferring resistance to the toxin.
鱼藤酮是一种广泛使用的杀虫剂,它能在大鼠中诱导出类似帕金森病的症状,并能在培养的多巴胺能神经元中导致死亡。虽然鱼藤酮是线粒体电子传递链复合体 I 的一种有效抑制剂,但它可以独立于复合体 I 抑制诱导多巴胺能神经元死亡。在这里,我们描述了裂殖酵母 Schizosaccharomyces pombe 中的鱼藤酮作用,该酵母缺乏复合体 I 并进行鱼藤酮不敏感的细胞呼吸。我们表明,鱼藤酮诱导处理的 S. pombe 细胞中活性氧 (ROS) 的产生以及线粒体网络的碎片化。虽然鱼藤酮对野生型 S. pombe 细胞的生长仅有适度的抑制作用,但对缺乏 ERK 型 MAP 激酶 Pmk1 或蛋白激酶 A (PKA) 的突变体的生长具有强烈的抑制作用。相比之下,缺乏 p38 MAP 激酶 Spc1 的细胞对鱼藤酮表现出适度的抗性。与这些发现一致,我们提供了证据表明 Pmk1 和 PKA,但不是 Spc1,是鱼藤酮处理的 S. pombe 细胞中清除 ROS 所必需的。我们的结果表明,裂殖酵母 S. pombe 可用于阐明鱼藤酮的复合体 I 非依赖性分子靶标以及赋予对毒素抗性的机制。