Annunen-Rasila Johanna, Ohlmeier Steffen, Tuokko Hanna, Veijola Johanna, Majamaa Kari
Department of Neurology, University of Oulu, Oulu, Finland.
Proteomics. 2007 Jun;7(13):2189-200. doi: 10.1002/pmic.200601031.
We have recently shown disorganization of the vimentin network in cultured cells deficient in oxidative phosphorylation (OXPHOS). We describe here the cellular responses to OXPHOS deficiency in osteosarcoma cells upon complex I (CI) and complex IV (CIV) inhibition, and upon the lack of mitochondrial DNA (rho0 cells). We examined the cytoskeletal organization and the distribution of mitochondria and analysed total proteome by 2-DE and vimentin expression by ELISA. Upon CIV inhibition and in rho0 cells, the vimentin network had collapsed around the nucleus and formed thick bundles. The mitochondria formed a perinuclear crescent upon CIV inhibition, whereas they accumulated around the nucleus in the rho0 cells, where the amount of vimentin was increased. Analysis of the total proteome revealed that a lack of mitochondrial DNA or inhibition of CI or CIV led to changes in the expression of cytoskeletal and cytoskeleton-associated proteins and proteins involved in apoptosis, OXPHOS, glycolysis, the tricarboxylic acid cycle, and oxidative stress responses. Our findings suggest that a deficiency in the energy converting system and oxidative stress can lead to cytoskeletal changes.
我们最近发现，在缺乏氧化磷酸化（OXPHOS）的培养细胞中，波形蛋白网络出现紊乱。在此，我们描述骨肉瘤细胞在复合体I（CI）和复合体IV（CIV）受到抑制以及缺乏线粒体DNA（rho0细胞）时对OXPHOS缺乏的细胞反应。我们检查了细胞骨架的组织情况以及线粒体的分布，并通过二维电泳分析了总蛋白质组，通过酶联免疫吸附测定法分析了波形蛋白的表达。在CIV受到抑制时以及在rho0细胞中，波形蛋白网络在细胞核周围坍塌并形成粗大的束状结构。在CIV受到抑制时，线粒体形成核周新月形结构，而在rho0细胞中，线粒体在细胞核周围聚集，其中波形蛋白的量增加。对总蛋白质组的分析表明，缺乏线粒体DNA或抑制CI或CIV会导致细胞骨架及细胞骨架相关蛋白以及参与细胞凋亡、OXPHOS、糖酵解、三羧酸循环和氧化应激反应的蛋白的表达发生变化。我们的研究结果表明，能量转换系统的缺陷和氧化应激可导致细胞骨架变化。
