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A genetic analysis of the response of Escherichia coli to cobalt stress

机译:大肠杆菌对钴胁迫反应的遗传分析

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Cobalt can be toxic and the way cells adapt to its presence is largely unknown. Here we carried out a transcriptomic analysis of Escherichia coli exposed to cobalt. A limited number of genes were either up- or downregulated. Upregulated genes include the isc and the nfuA genes encoding Fe/S biogenesis assisting factors, and the rcnA gene encoding a cobalt efflux system. Downregulated genes are implicated in anaerobic metabolism (narK, nirB, hybO, grcA), metal transport (feoB, nikA), sulfate/thiosulfate import (cysP), and one is of unknown function (yeeE). Cobalt regulation of isc, nfuA, hybO, cysP and yeeE genes was found to involve IscR, a Fe/S transcriptional regulator. Previously, the Suf Fe/S biogenesis machinery was found to be important for cobalt stress adaptation, but suf genes did not show up in the microarray analysis. Therefore, we used qRT-PCR analysis and found that cobalt induced the suf operon expression. Moreover, kinetic analysis of the cobalt-mediated induction of the suf operon expression allowed us to propose that cobalt toxicity is caused first by impaired Fe/S biogenesis, followed by decreased iron bioavailability and eventually oxidative stress.
机译:钴可能是有毒的,并且细胞对其存在的适应方式尚不清楚。在这里,我们对暴露于钴的大肠杆菌进行了转录组学分析。有限数量的基因被上调或下调。上调的基因包括编码Fe / S生物合成辅助因子的isc和nfuA基因,以及编码钴外排系统的rcnA基因。下调的基因与厌氧代谢(narK,nirB,hybO,grcA),金属转运(feoB,nikA),硫酸盐/硫代硫酸盐进口(cysP)有关,其中一个功能未知(yeeE)。发现isc,nfuA,hybO,cysP和yeeE基因的钴调节涉及Fe / S转录调节剂IscR。以前,发现Suf Fe / S生物发生机制对钴胁迫适应性很重要,但是在微阵列分析中并未显示suf基因。因此,我们使用qRT-PCR分析,发现钴诱导了suf操纵子表达。此外,对钴介导的suf操纵子表达的动力学分析使我们提出,钴毒性首先是由Fe / S生物发生受损引起,然后是铁生物利用度降低,最终是氧化应激。

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