Impact of rpoS Deletion on the Proteome of Escherichia coli Grown Planktonically and as Biofilm
Impact of rpoS Deletion on the Proteome of Escherichia coli Grown Planktonically and as Biofilm
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DOI：
10.1021/pr8001723
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发表时间：
2008-11-01
影响因子：
4.4
通讯作者：
Jouenne, Thierry
中科院分区：
文献类型：
--
作者：
Collet, Anthony;Cosette, Pascal;Jouenne, Thierry
为了研究rpos在以生物膜形式生长的大肠杆菌细胞基因表达中的作用，我们比较了rpos突变体和野生型菌株的蛋白质组。实验是在浮游细胞(指数生长期或平稳生长期)和在玻璃棉上形成的生物膜上进行的。从生物膜和浮游野生生物获得的凝胶的逐点比较表明，22%至30%的检测斑点的强度受到生长模式的影响，具体取决于所使用的对照。主成分分析用于解释蛋白质斑点密度的变化，将指数期细胞(野生型和突变型)与其他培养条件和次要的72岁培养细胞区分开来。统计分析表明，rpos突变并没有显著改变指数生长期细胞的蛋白质组，这种差异只涉及3%的蛋白质组。然而，将孵育时间从8h增加到72h，显著增加了变化的蛋白质数量。聚类分析表明，rpos在生物膜细胞基因表达的特殊性中发挥了作用，但低于稳定期细菌。我们鉴定了35个受rpos调控的蛋白质，这些蛋白质已经或没有被描述为受这个西格玛因子控制。对于其中一些细菌，rpos的调节方式明显依赖于培养条件(浮游生物与生物膜)。
To investigate the role of rpoS in gene expression of Escherichia coli cells grown as biofilms, we compared the proteomes of a rpoS mutant and the wild-type strain. Experiments were performed on planktonic cells (in exponential or stationary growth phase) and biofilms developed on glass wool. Spot-by-spot comparison of gels obtained from biofilm and planktonic wild-type organisms showed that the intensity of between 22 and 30% of detected spots was affected by the growth mode, depending of the control used. Principal component analysis, used to interpret the variations in protein spot densities, discriminated exponential-phase cells (wild-type and mutant) from the other incubation conditions and secondarily 72-old cultures. The statistical analysis demonstrated that the rpoS mutation did not significantly modify the proteome of exponential-growth phase cells, the differences involving only 3% of the proteome. However, increasing the incubation time from 8 to 72 h noticeably increased the number of changed proteins. A cluster analysis showed that RpoS plays a role in the special nature of the gene expression of biofilm cells but lower than in stationary-phase bacteria. We identified 35 rpoS-regulated proteins that were already or not described as controlled by this sigma factor. For some of them, the mode of regulation by RpoS was obviously dependent on the culture condition (planktonic vs biofilm).