首页> 美国卫生研究院文献>Journal of Bacteriology >Pseudomonas aeruginosa LysR PA4203 Regulator NmoR Acts as a Repressor of the PA4202 nmoA Gene Encoding a Nitronate Monooxygenase
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Pseudomonas aeruginosa LysR PA4203 Regulator NmoR Acts as a Repressor of the PA4202 nmoA Gene Encoding a Nitronate Monooxygenase

机译:铜绿假单胞菌LysR PA4203调节剂NmoR充当PA4202 nmoA基因的阻遏物编码亚硝酸单加氧酶

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摘要

The PA4203 gene encodes a LysR regulator and lies between the ppgL gene (PA4204), which encodes a periplasmic gluconolactonase, and, in the opposite orientation, the PA4202 (nmoA) gene, coding for a nitronate monooxygenase, and ddlA (PA4201), encoding a d-alanine alanine ligase. The intergenic regions between PA4203 and ppgL and between PA4203 and nmoA are very short (79 and 107 nucleotides, respectively). Here we show that PA4203 (nmoR) represses its own transcription and the expression of nmoA. A chromatin immunoprecipitation analysis showed the presence of a single NmoR binding site between nmoA and nmoR, which was confirmed by electrophoretic mobility shift assays (EMSAs) with the purified NmoR protein. Despite this observation, a transcriptome analysis revealed more genes to be affected in an nmoR mutant, including genes known to be part of the MexT LysR activator regulon. The PA1225 gene, encoding a quinone oxidoreductase, was the most highly upregulated gene in the nmoR deletion mutant, independently of MexT. Finally, deletion of the nmoA gene resulted in an increased sensitivity of the cells to 3-nitropropionic acid (3-NPA), confirming the role of the nitronate monooxygenase protein in the detoxification of nitronate.
机译:PA4203基因编码LysR调节剂,位于ppgL基因(PA4204)和ddlA(PA4201)之间,ppgL基因(PA4204)编码周质葡萄糖酸内酯酶,PA4202(nmoA)基因编码亚硝酸单加氧酶,而ddlA(PA4201)编码d-丙氨酸丙氨酸连接酶。 PA4203和ppgL之间以及PA4203和nmoA之间的基因间区域非常短(分别为79和107个核苷酸)。在这里,我们显示PA4203(nmoR)抑制其自身的转录和nmoA的表达。染色质免疫沉淀分析显示nmoA和nmoR之间存在单个NmoR结合位点,这已通过纯化的NmoR蛋白的电泳迁移率变动分析(EMSA)得以证实。尽管有这一观察结果,转录组分析显示nmoR突变体中有更多基因受到影响,包括已知是MexT LysR激活剂调节子一部分的基因。编码醌氧化还原酶的PA1225基因是nmoR缺失突变体中上调程度最高的基因,独立于MexT。最后,nmoA基因的缺失导致细胞对3-硝基丙酸(3-NPA)的敏感性增加,从而证实了亚硝酸盐单加氧酶蛋白在亚硝酸盐解毒中的作用。

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