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Genetics and proteomics analyses reveal the roles of PhoB1 and PhoB2 regulators in bacterial responses to arsenite and phosphate.
Research in Microbiology ( IF 2.6 ) Pub Date : 2019-07-03 , DOI: 10.1016/j.resmic.2019.06.006
Zixu Qiao 1 , Jing Huang 1 , Yajing Cao 1 , Kaixiang Shi 1 , Gejiao Wang 1
Affiliation  

In bacteria, phosphate (Pi) stress response is governed by the two-component regulatory system, sensor kinase PhoR and its cognate response regulatory protein PhoB. The arsenite [As(III)]-oxidizing bacterium Agrobacterium tumefaciens GW4 contains two phoB genes, phoB1 and phoB2. phoB1 is adjacent to As(III)-oxidizing genes, however, the functions of PhoB1 and PhoB2 remain unclear. Here, phoB1 and phoB2 were each deleted in-frame, and proteomics, qRT-PCR and protein-DNA interaction were performed. We found that (1) phoB1 and phoB2 were both upregulated under low Pi conditions and phoB1 was induced by As(III), but phoB2 was not; (2) deletion of phoB1 reduced As(III)-oxidizing efficiency and protein-DNA interaction analysis showed PhoB1 could interact with aioXSR promoter to regulate As(III) oxidation; (3) deletions of phoB1 or phoB2 both reduced exopolysaccharides (EPS) synthesis; and (4) PhoB1 influenced Pi uptake, As(III) oxidation, EPS synthesis, TCA cycle, energy production and stress response with As(III), and PhoB2 was associated with Pi uptake and EPS synthesis in low Pi conditions. These results showed PhoB1 and PhoB2 were both involved in Pi acquisition, PhoB1 was more important with As(III) and PhoB2 played a major role without As(III). Strain GW4 uses these two regulators to survive under low Pi and arsenic-rich environments.

中文翻译:

遗传学和蛋白质组学分析揭示了PhoB1和PhoB2调节剂在细菌对亚砷酸盐和磷酸盐的反应中的作用。

在细菌中,磷酸盐(Pi)应激反应受两组分调节系统即传感器激酶PhoR及其同源反应调节蛋白PhoB的控制。砷化物[As(III)]-氧化细菌根癌农杆菌GW4包含两个phoB基因phoB1和phoB2。phoB1与As(III)氧化基因相邻,但是,PhoB1和PhoB2的功能仍不清楚。在这里,phoB1和phoB2分别在读框内被删除,并进行了蛋白质组学,qRT-PCR和蛋白质-DNA相互作用。我们发现(1)在低Pi条件下phoB1和phoB2均被上调,As(III)诱导phoB1,而phoB2则没有。(2)phoB1的缺失降低了As(III)的氧化效率,蛋白质-DNA相互作用分析表明PhoB1可以与aioXSR启动子相互作用来调节As(III)的氧化。(3)phoB1或phoB2的缺失均降低了胞外多糖(EPS)的合成;(4)PhoB1影响Pi吸收,As(III)氧化,EPS合成,TCA循环,As(III)的能量产生和应力响应,而PhoB2与低Pi条件下的Pi吸收和EPS合成相关。这些结果表明,PhoB1和PhoB2都参与了Pi的获取,PhoB1在As(III)中更重要,而PhoB2在没有As(III)时起主要作用。GW4菌株使用这两个调节剂在低Pi和富含砷的环境中生存。这些结果表明,PhoB1和PhoB2都参与了Pi的获取,PhoB1在As(III)中更重要,而PhoB2在没有As(III)时起主要作用。GW4菌株使用这两个调节剂在低Pi和富含砷的环境中生存。这些结果表明,PhoB1和PhoB2都参与了Pi的获取,PhoB1在As(III)中更重要,而PhoB2在没有As(III)时起主要作用。GW4菌株使用这两个调节剂在低Pi和富含砷的环境中生存。
更新日期:2019-11-01
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