Zhang Dawei, Lu Chao, Whiteman Matthew, Chance Britton, Armstrong Jeffrey S
Department of Biochemistry, National University of Singapore, Singapore 117597, Singapore; Institute of Bioengineering and Nanotechnology, Singapore 138669, Singapore.
Department of Biochemistry, National University of Singapore, Singapore 117597, Singapore.
J Biol Chem. 2008 Feb 8;283(6):3476-3486. doi: 10.1074/jbc.M707528200. Epub 2007 Dec 5.
The role of the mitochondrial permeability transition (MPT) in apoptosis and necrosis is controversial. Here we show that the MPT regulates the release of cytochrome c for apoptosis during endoplasmic reticulum (ER) stress by remodeling the cristae junction (CJ). CEM cells, HCT116 colon cancer cells, and murine embryo fibroblast cells were treated with the ER stressor thapsigargin (THG), which led to cyclophilin D-dependent mitochondrial release of the profusion GTPase optic atrophy 1 (OPA1), which controls CJ integrity, and cytochrome c, leading to apoptosis. Interference RNA knockdown of Bax blocked OPA1 and cytochrome c release after THG treatment but did not prevent the MPT, showing that Bax was essential for the release of cytochrome c by MPT. In isolated mitochondria, MPT led to OPA1 and cytochrome c release independently of voltage-dependent anion channel and the outer membrane, indicating that the MPT is an inner membrane phenomenon. Last, the MPT was regulated by the electron transport chain but not mitochondrial reactive oxygen species, since THG-induced cell death was not blocked by antioxidants and did not occur in cells lacking mitochondrial DNA. Our results show that the MPT regulates CJ remodeling for cytochrome c-dependent apoptosis induced by ER stress and that mitochondrial electron transport is indispensable for this process.
线粒体通透性转换(MPT)在细胞凋亡和坏死中的作用存在争议。在此我们表明,MPT通过重塑嵴连接(CJ)来调节内质网(ER)应激期间细胞色素c的释放以引发凋亡。用ER应激剂毒胡萝卜素(THG)处理人急性淋巴细胞白血病细胞(CEM细胞)、人结肠癌细胞系HCT116以及小鼠胚胎成纤维细胞,这导致亲环素D依赖性的线粒体释放控制CJ完整性的线粒体融合GTP酶视神经萎缩蛋白1(OPA1)以及细胞色素c,从而引发凋亡。干扰RNA敲低Bax可阻断THG处理后的OPA1和细胞色素c释放,但不能阻止MPT,这表明Bax对于MPT介导的细胞色素c释放至关重要。在分离的线粒体中,MPT导致OPA1和细胞色素c释放,且不依赖于电压依赖性阴离子通道和外膜,这表明MPT是一种内膜现象。最后,MPT受电子传递链调控而非线粒体活性氧调控,因为THG诱导的细胞死亡未被抗氧化剂阻断,且在缺乏线粒体DNA的细胞中未发生。我们的结果表明,MPT调节ER应激诱导的依赖细胞色素c的凋亡过程中的CJ重塑,并且线粒体电子传递对于该过程不可或缺。