Effect of Lon protease knockdown on mitochondrial function in HeLa cells
Effect of Lon protease knockdown on mitochondrial function in HeLa cells
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DOI：
10.1016/j.biochi.2013.12.005
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发表时间：
2014-05-01
期刊：
影响因子：
3.9
通讯作者：
Bulteau, Anne-Laure
中科院分区：
文献类型：
--
作者：
Bayot, Aurelien;Gareil, Monique;Bulteau, Anne-Laure
ATP依赖性蛋白酶目前正在成为线粒体功能的关键调节因子。在这些蛋白水解系统中，Lon蛋白酶参与控制线粒体基质中的选择性蛋白质周转。在Lon不存在的情况下，酵母细胞已被证明在基质空间中积累电子致密的包涵体，使线粒体基因组的完整性松散，并且是呼吸缺陷的。为了解决Lon在人细胞中线粒体功能中的作用，我们建立了用表达在多西环素诱导的启动子控制下的shRNA的载体稳定转染的HeLa细胞系。我们已经证明，Lon蛋白酶的减少导致该细胞系中的温和表型，与在其他细胞类型如WI-38成纤维细胞中观察到的相反。然而，Lon蛋白酶的缺乏导致ROS产生的增加和羰基化蛋白在线粒体中的积累。我们的研究表明，Lon蛋白酶具有多种靶点，并且可能根据细胞类型发挥不同的作用。(C)2013年Elsevier Masson SAS。All rights reserved.
ATP-dependent proteases are currently emerging as key regulators of mitochondrial functions. Among these proteolytic systems, Lon protease is involved in the control of selective protein turnover in the mitochondrial matrix. In the absence of Lon, yeast cells have been shown to accumulate electron-dense inclusion bodies in the matrix space, to loose integrity of mitochondrial genome and to be respiratory deficient. In order to address the role of Lon in mitochondrial functionality in human cells, we have set up a HeLa cell line stably transfected with a vector expressing a shRNA under the control of a promoter which is inducible with doxycycline. We have demonstrated that reduction of Lon protease results in a mild phenotype in this cell line in contrast with what have been observed in other cell types such as WI-38 fibroblasts. Nevertheless, deficiency in Lon protease led to an increase in ROS production and to an accumulation of carbonylated protein in the mitochondria. Our study suggests that Lon protease has a wide variety of targets and is likely to play different roles depending of the cell type. (C) 2013 Elsevier Masson SAS. All rights reserved.