GadE regulates fliC gene transcription and motility in Escherichia coli.

William R Schwan, Nicole L Flohr, Abigail R Multerer, Jordan C Starkey
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引用次数: 4

Abstract

Background: Escherichia coli (E. coli) express flagella to ascend human urinary tracts. To survive in the acidic pH of human urine, E. coli uses the glutamate decarboxylase acid response system, which is regulated by the GadE protein.

Aim: To determine if growth in an acidic pH environment affected fliC transcription and whether GadE regulated that transcription.

Methods: A fliC-lacZ reporter fusion was created on a single copy number plasmid to assess the effects of acidic pH on fliC transcription. Further, a ΔgadE mutant strain of a uropathogenic E. coli was created and tested for motility compared to the wild-type strain.

Results: Escherichia coli cells carrying the fliC-lacZ fusion displayed significantly less fliC transcription when grown in an acidic pH medium compared to when grown in a neutral pH medium. Transcription of fliC fell further when the E. coli was grown in an acidic pH/high osmolarity environment. Since GadE is a critical regulator of one acid response system, fliC transcription was tested in a gadE mutant strain grown under acidic conditions. Expression of fliC was derepressed in the E. coli gadE mutant strain grown under acidic conditions compared to that in wild-type bacteria under the same conditions. Furthermore, a gadE mutation in a uropathogenic E. coli background exhibited significantly greater motility than the wild-type strain following growth in an acidic medium.

Conclusion: Together, our results suggest that GadE may down-regulate fliC transcription and motility in E. coli grown under acidic conditions.

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GadE调节大肠杆菌中fliC基因的转录和运动。
背景:大肠杆菌(E. coli)表达鞭毛,可沿人类尿路上升。为了在人类尿液的酸性pH中生存,大肠杆菌使用谷氨酸脱羧酶酸反应系统,该系统由GadE蛋白调节。目的:确定在酸性pH环境下生长是否影响fliC转录,以及GadE是否调节这种转录。方法:在单拷贝数质粒上构建fliC- lacz报告基因融合,评估酸性pH对fliC转录的影响。此外,建立了一种ΔgadE尿路致病性大肠杆菌突变株,并与野生型菌株进行了活力测试。结果:携带fliC- lacz融合的大肠杆菌细胞在酸性pH培养基中生长时,与在中性pH培养基中生长时相比,fliC转录明显减少。当大肠杆菌生长在酸性pH/高渗透压环境中时,fliC的转录进一步下降。由于GadE是一种酸反应系统的关键调节因子,因此在酸性条件下生长的GadE突变株中测试了fliC的转录。与野生型细菌相比,在酸性条件下生长的大肠杆菌gadE突变株中fliC的表达被抑制。此外,尿路致病性大肠杆菌背景的gadE突变在酸性培养基中生长后,表现出比野生型菌株更大的运动性。结论:在酸性条件下生长的大肠杆菌中,GadE可能下调fliC的转录和活性。
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