Transposon-Directed Insertion-Site Sequencing Reveals Glycolysis Gene gpmA as Part of the H(2)O(2) Defense Mechanisms in Escherichia coli.
Transposon-Directed Insertion-Site Sequencing Reveals Glycolysis Gene gpmA as Part of the H(2)O(2) Defense Mechanisms in Escherichia coli.
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DOI：
10.3390/antiox11102053
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发表时间：
2022-10-18
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影响因子：
--
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中科院分区：
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过氧化氢（H2O2）是病原性感染防御机制的常见效应物。然而，涉及H2O2耐受性的细菌因素仍不清楚。在这里，我们使用转座子定向插入位点测序（TraDIS），一种允许筛选全基因组的技术，以确定大肠杆菌中涉及H2O2耐受性的基因。我们的TraDIS分析鉴定了10个在H2O2暴露下具有适应性缺陷的突变体，其中先前与H2O2相关的基因（oxyR、dps、dksA、rpoS、hfq和polA）和其他与H2O2耐受性没有已知关联的基因。coli（corA、rbsR、nhaA和gpmA）。这是第一次描述糖酵解相关基因gpmA对大肠杆菌易感性的影响。coli转化为H2O2。事实上，验证性实验表明，gpmA的删除导致了一个特定的超敏反应H2O2的主要H2O2清除剂基因katG的删除。这种超敏反应不是由于过氧化氢酶功能的改变，并且与碳源或氧的存在无关。在H2O2暴露下gpmA的转录上调，突出了其在氧化应激下的作用。总之，我们的TraDIS方法将gpmA鉴定为大肠杆菌中氧化应激防御机制的成员。杆菌
Hydrogen peroxide (H2O2) is a common effector of defense mechanisms against pathogenic infections. However, bacterial factors involved in H2O2 tolerance remain unclear. Here we used transposon-directed insertion-site sequencing (TraDIS), a technique allowing the screening of the whole genome, to identify genes implicated in H2O2 tolerance in Escherichia coli. Our TraDIS analysis identified 10 mutants with fitness defect upon H2O2 exposure, among which previously H2O2-associated genes (oxyR, dps, dksA, rpoS, hfq and polA) and other genes with no known association with H2O2 tolerance in E. coli (corA, rbsR, nhaA and gpmA). This is the first description of the impact of gpmA, a gene involved in glycolysis, on the susceptibility of E. coli to H2O2. Indeed, confirmatory experiments showed that the deletion of gpmA led to a specific hypersensitivity to H2O2 comparable to the deletion of the major H2O2 scavenger gene katG. This hypersensitivity was not due to an alteration of catalase function and was independent of the carbon source or the presence of oxygen. Transcription of gpmA was upregulated under H2O2 exposure, highlighting its role under oxidative stress. In summary, our TraDIS approach identified gpmA as a member of the oxidative stress defense mechanism in E. coli.
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影响因子：
34.3
作者：
Anderson, John D.;Bagamian, Karoun H.;Rheingans, Richard
通讯作者：
Rheingans, Richard
大肠杆菌K-12的构造框架，单基因敲除突变体：Keio Collection。
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
14.9
作者：
Keseler IM;Collado-Vides J;Santos-Zavaleta A;Peralta-Gil M;Gama-Castro S;Muñiz-Rascado L;Bonavides-Martinez C;Paley S;Krummenacker M;Altman T;Kaipa P;Spaulding A;Pacheco J;Latendresse M;Fulcher C;Sarker M;Shearer AG;Mackie A;Paulsen I;Gunsalus RP;Karp PD
通讯作者：
Karp PD
影响因子：
3.6
作者：
Henard CA;Tapscott T;Crawford MA;Husain M;Doulias PT;Porwollik S;Liu L;McClelland M;Ischiropoulos H;Vázquez-Torres A
通讯作者：
Vázquez-Torres A
影响因子：
10.5
作者：
Imlay JA
通讯作者：
Imlay JA