The Ohio State University, Department of Chemistry, 100 West 18th Avenue, Columbus, OH 43210, USA.
J Bacteriol. 2010 Apr;192(8):2193-209. doi: 10.1128/JB.00484-09. Epub 2010 Jan 8.
YidC depletion affects membrane protein insertion and leads to a defect in the growth of the Escherichia coli cell. We analyzed global changes in gene expression upon YidC depletion to determine the importance of YidC for cellular functions using a gene chip method to compare the transcriptomes of JS71 (control) and JS7131 (yidC depletion strain). Of the more than 4,300 genes identified, 163 were upregulated and 99 were downregulated upon YidC depletion, including genes which are responsible for DNA/RNA repair; energy metabolism; various transporters, proteases and chaperones; stress response; and translation and transcription functions. Real-time PCR was performed on selected genes to confirm the results. Specifically, we found upregulation of the genes encoding the energy transduction proteins F(1)F(o) ATP synthase and cytochrome bo(3) oxidase due to perturbation in assembly when YidC was depleted. We also determined that the high-level induction of the PspA stress protein under YidC depletion conditions is roughly 10-fold higher than the activation due to the addition of protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), which dissipates the proton motive force. In addition, the gene chip data reveal the Cpx stress pathway is activated upon YidC depletion. The data show the broad physiological contribution of YidC to the bacterial cell and the considerable ramification to the cell when it is depleted.
YidC 缺失会影响膜蛋白插入,并导致大肠杆菌细胞生长缺陷。我们使用基因芯片方法分析了 YidC 缺失后基因表达的全局变化,以确定 YidC 对细胞功能的重要性,从而比较 JS71(对照)和 JS7131(yidC 缺失菌株)的转录组。在鉴定出的 4300 多个基因中,有 163 个基因上调,99 个基因下调,其中包括负责 DNA/RNA 修复的基因;能量代谢;各种转运蛋白、蛋白酶和伴侣蛋白;应激反应;以及翻译和转录功能。我们对选定的基因进行了实时 PCR 以验证结果。具体来说,我们发现当 YidC 缺失时,由于组装受到干扰,编码能量转导蛋白 F(1)F(o)ATP 合酶和细胞色素 bo(3)氧化酶的基因上调。我们还确定,在 YidC 缺失条件下,PspA 应激蛋白的高水平诱导比添加质子载体羰基氰化物 m-氯苯腙(CCCP)引起的激活高约 10 倍,CCCP 会耗散质子动力势。此外,基因芯片数据显示 Cpx 应激途径在 YidC 缺失时被激活。数据显示了 YidC 对细菌细胞的广泛生理贡献,以及当它缺失时对细胞的重大影响。