The OxyR-regulated phnW gene encoding 2-aminoethylphosphonate:pyruvate aminotransferase helps protect Pseudomonas aeruginosa from tert-butyl hydroperoxide.
The OxyR-regulated phnW gene encoding 2-aminoethylphosphonate:pyruvate aminotransferase helps protect Pseudomonas aeruginosa from tert-butyl hydroperoxide.
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DOI：
10.1371/journal.pone.0189066
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Hassett DJ
中科院分区：
文献类型：
--
作者：
Panmanee W;Charoenlap N;Atichartpongkul S;Mahavihakanont A;Whiteside MD;Winsor G;Brinkman FSL;Mongkolsuk S;Hassett DJ
细菌反激活因子的LysR成员OxyR控制着参与革兰氏阴性病原体铜绿假单胞菌对H2O2和有机（烷基）氢过氧化物（AHP）反应的基因的转录。我们之前的研究表明，在浮游细菌和生物膜细菌中，缺乏氧还原的生物分别被<2或500 mM的H2O2迅速杀死。在这项研究中，我们首先采用生物信息学方法，通过扫描整个P. aeruginosa PAO1基因组，寻找典型的OxyR启动子识别序列（ATAG-N7-CTAT-N7-ATAG-N7-CTAT）来阐明OxyR的潜在调控广度。在bb100个潜在的OxyR控制基因中，策略性地选择了40个不预测直接参与氧化应激反应的基因（如过氧化氢酶、过氧化物酶等），并通过RT-PCR分析筛选这些基因，以进行潜在的OxyR阳性或阴性控制。当将oxyR突变体与PAO1表达进行比较时，发现40个基因中有7个存在差异，ß-半乳糖苷酶报告基因试验证实了这一点。其中，编码2-氨基乙基膦酸：丙酮酸转氨酶的phnW在oxyR突变体中的表达比野生型细菌低。电泳迁移位移分析表明OxyR与phnW启动子结合，dna酶I足迹分析也揭示了OxyR与phnW启动子结合的序列。有趣的是，phnW突变体比野生型细菌更容易受到t-丁基过氧化氢（t-BOOH）处理。虽然我们无法确定这种现象的直接机制，但我们认为这可能是由于phnW突变体中AHP基因转录的调节导致该菌株相对于野生型生物降解t-BOOH的效率降低。
The LysR member of bacterial transactivators, OxyR, governs transcription of genes involved in the response to H2O2 and organic (alkyl) hydroperoxides (AHP) in the Gram-negative pathogen, Pseudomonas aeruginosa. We have previously shown that organisms lacking OxyR are rapidly killed by <2 or 500 mM H2O2 in planktonic and biofilm bacteria, respectively. In this study, we first employed a bioinformatic approach to elucidate the potential regulatory breadth of OxyR by scanning the entire P. aeruginosa PAO1 genome for canonical OxyR promoter recognition sequences (ATAG-N7-CTAT-N7-ATAG-N7-CTAT). Of >100 potential OxyR-controlled genes, 40 were strategically selected that were not predicted to be involved in the direct response to oxidative stress (e.g., catalase, peroxidase, etc.) and screened such genes by RT-PCR analysis for potentially positive or negative control by OxyR. Differences were found in 7 of 40 genes when comparing an oxyR mutant vs. PAO1 expression that was confirmed by ß-galactosidase reporter assays. Among these, phnW, encoding 2-aminoethylphosphonate:pyruvate aminotransferase, exhibited reduced expression in the oxyR mutant compared to wild-type bacteria. Electrophoretic mobility shift assays indicated binding of OxyR to the phnW promoter and DNase I footprinting analysis also revealed the sequences to which OxyR bound. Interestingly, a phnW mutant was more susceptible to t-butyl-hydroperoxide (t-BOOH) treatment than wild-type bacteria. Although we were unable to define the direct mechanism underlying this phenomenon, we believe that this may be due to a reduced efficiency for this strain to degrade t-BOOH relative to wild-type organisms because of modulation of AHP gene transcription in the phnW mutant.
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DOI：
10.1111/j.1432-1033.1983.tb07436.x
发表时间：
1983-01-01
期刊：
EUROPEAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
DUMORA, C;LACOSTE, AM;CASSAIGNE, A
通讯作者：
CASSAIGNE, A
影响因子：
64.5
作者：
TOLEDANO, MB;KULLIK, I;STORZ, G
通讯作者：
STORZ, G
DOI：
10.1126/science.1215643
发表时间：
2012-04-27
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Seth D;Hausladen A;Wang YJ;Stamler JS
通讯作者：
Stamler JS
影响因子：
3.5
作者：
SCHWEIZER, HP;HOANG, TT
通讯作者：
HOANG, TT
影响因子：
4.8
作者：
Borisova, Svetlana A.;Christman, Harry D.;Metcalf, William W.
通讯作者：
Metcalf, William W.