Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1085.
Department of Neurology, University of Michigan School of Medicine, Ann Arbor, MI 48109-1085.
Mol Biol Cell. 2020 Aug 1;31(17):1931-1942. doi: 10.1091/mbc.E20-01-0063. Epub 2020 Jun 17.
Reactive oxygen species (ROS)-induced oxidative stress has been associated with diseases such as amyotrophic lateral sclerosis, stroke, and cancer. While the effect of ROS on mitochondria and endoplasmic reticulum (ER) has been well documented, its consequence on the Golgi apparatus is less well understood. In this study, we characterized the Golgi structure and function in HeLa cells after exposure to hydrogen peroxide (HO), a reagent commonly used to introduce ROS to cells. Treatment of cells with 1 mM HO for 10 min resulted in the degradation of Arl1 and dissociation of GRIP domain-containing proteins Golgin-97 and Golgin-245 from the -Golgi. This effect could be rescued by treatment of cells with a ROS scavenger -acetyl cysteine or protease inhibitors. Structurally, HO treatment reduced the number of cisternal membranes per Golgi stack, suggesting a loss of -Golgi cisternae. Functionally, HO treatment inhibited both anterograde and retrograde protein transport, consistent with the loss of membrane tethers on the -Golgi cisternae. This study revealed membrane tethers at the -Golgi as novel specific targets of ROS in cells.
活性氧(ROS)诱导的氧化应激与肌萎缩侧索硬化症、中风和癌症等疾病有关。虽然 ROS 对线粒体和内质网(ER)的影响已经得到很好的证明,但它对高尔基体的影响却知之甚少。在这项研究中,我们在 HeLa 细胞中研究了暴露于过氧化氢(HO)后高尔基体的结构和功能,HO 是一种常用于向细胞中引入 ROS 的试剂。用 1mMHO 处理细胞 10 分钟会导致 Arl1 的降解和 GRIP 结构域蛋白 Golgin-97 和 Golgin-245 从 -高尔基体解离。这种作用可以通过用 ROS 清除剂 -乙酰半胱氨酸或蛋白酶抑制剂处理细胞来挽救。从结构上看,HO 处理减少了每个高尔基体堆叠的膜室数量,表明 -高尔基体膜室的丢失。从功能上看,HO 处理抑制了顺行和逆行蛋白运输,与 -高尔基体膜室上的膜连接蛋白丢失一致。这项研究揭示了 -高尔基体上的膜连接蛋白是细胞中 ROS 的新型特定靶标。