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Activation of glucose transport under oxidative stress in Escherichia coli

机译:大肠杆菌中氧化应激下葡萄糖转运的激活

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摘要

Global transcription studies have identified a large number of redox-responsive genes, although the biological relevance of this regulation has not been experimentally tested. In particular, several genes coding for enzymes involved in glucose metabolism have been identified as redox-responsive in Escherichia coli. However, only zwf, which codes for glucose-6-phosphate dehydrogenase, has been shown experimentally to affect the cellular resistance to oxidative stress. We addressed the question of whether ptsG, coding for the membrane component of the glucose-specific transporter system, and pgi, coding for phosphoglucose isomerase, two additional genes identified in whole-genome functional screens, are indeed relevant in antioxidant defense. PTS assays showed that glucose transport was induced under oxidative stress elicited by the superoxide-producing agent paraquat (PQ). This induction of glucose transport under oxidative stress was dependent on the soxRS genes, coding for a sensor- transcriptional activator system, and ptsG. The binding of purified SoxS to the ptsG promoter region was shown by gel mobility-shift assay, and the activation of the ptsG promoter P1 was demonstrated by primer extension assays. Finally, a ptsG mutant strain was hypersensitive to PQ when grown in rich medium plus glucose, but not in rich medium without glucose. The pgi gene showed the same pattern of regulation by oxidative stress under the control of the SoxRS system, and a strain carrying a pgi deletion was hypersensitive to PQ.
机译:尽管尚未对该实验的生物学相关性进行过测试,但全球转录研究已经鉴定出了大量的氧化还原反应基因。特别是,已鉴定出几种编码参与葡萄糖代谢的酶的基因在大肠杆菌中具有氧化还原反应性。然而,只有zwf编码6-磷酸葡萄糖磷酸脱氢酶,已通过实验证明会影响细胞对氧化应激的抵抗力。我们解决了以下问题:在全基因组功能筛选中鉴定出的另外两个基因,编码葡萄糖特异性转运系统膜成分的ptsG和编码磷酸葡萄糖异构酶的pgi是否确实与抗氧化剂防御有关。 PTS分析表明,葡萄糖运输是在超氧化物产生剂百草枯(PQ)引起的氧化应激下诱导的。在氧化应激下对葡萄糖转运的这种诱导取决于编码传感器转录激活系统和ptsG的soxRS基因。纯化的SoxS与ptsG启动子区域的结合通过凝胶迁移率移动分析显示,而ptsG启动子P1的激活通过引物延伸分析证明。最后,当在富含葡萄糖的培养基中生长时,ptsG突变株对PQ敏感,而在没有葡萄糖的富含培养基中则不敏感。 pgi基因在SoxRS系统的控制下表现出相同的氧化应激调控模式,带有pgi缺失的菌株对PQ敏感。

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  • 来源
    《Archives of Microbiology》 |2008年第1期|41-49|共9页
  • 作者单位

    Department of Microbiology Morrill Science Center 4 North University of Massachusetts Amherst MA 01003 USA;

    Department of Microbiology Morrill Science Center 4 North University of Massachusetts Amherst MA 01003 USA;

    Department of Microbiology Morrill Science Center 4 North University of Massachusetts Amherst MA 01003 USA;

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