Exposure of E. coli to DNA-Methylating Agents Impairs Biofilm Formation and Invasion of Eukaryotic Cells via Down Regulation of the N-Acetylneuraminate Lyase NanA.
Exposure of E. coli to DNA-Methylating Agents Impairs Biofilm Formation and Invasion of Eukaryotic Cells via Down Regulation of the N-Acetylneuraminate Lyase NanA.
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DOI：
10.3389/fmicb.2016.00147
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发表时间：
2016
影响因子：
5.2
通讯作者：
Duilio A
中科院分区：
文献类型：
--
作者：
Di Pasquale P;Caterino M;Di Somma A;Squillace M;Rossi E;Landini P;Iebba V;Schippa S;Papa R;Selan L;Artini M;Palamara AT;Duilio A
DNA甲基化损伤可以由内源和外源化学物质引起，这使得每一个活着的有机体都制定了合适的反应策略。我们用差异蛋白质组学的方法研究了烷基化试剂甲基甲烷磺酸(MMS)对大肠杆菌蛋白质表达的影响。定量蛋白质组学数据显示，属于糖酵解途径的酶和脂肪酸降解大量下调，强烈表明能量产生减少。参与唾液酸代谢的N-乙酰神经氨酸裂解酶(NANA)的表达也显著降低。利用缺失的nana突变体和底物类似物Dana作为竞争抑制物，我们证明了nana的下调影响了E.coliMV1161的生物膜形成和黏附特性。接触烷化剂也减少了粘附性侵袭性大肠杆菌(AIEC)菌株LF82的生物膜形成和细菌对Caco-2真核细胞系的粘附性。我们的数据表明，甲基化应激损害了大肠杆菌的粘附性，并提示NaNA在生物膜形成和细菌宿主相互作用中可能发挥作用。
DNA methylation damage can be induced by endogenous and exogenous chemical agents, which has led every living organism to develop suitable response strategies. We investigated protein expression profiles of Escherichia coli upon exposure to the alkylating agent methyl-methane sulfonate (MMS) by differential proteomics. Quantitative proteomic data showed a massive downregulation of enzymes belonging to the glycolytic pathway and fatty acids degradation, strongly suggesting a decrease of energy production. A strong reduction in the expression of the N-acetylneuraminate lyases (NanA) involved in the sialic acid metabolism was also observed. Using a null NanA mutant and DANA, a substrate analog acting as competitive inhibitor, we demonstrated that down regulation of NanA affects biofilm formation and adhesion properties of E. coli MV1161. Exposure to alkylating agents also decreased biofilm formation and bacterial adhesion to Caco-2 eukaryotic cell line by the adherent invasive E. coli (AIEC) strain LF82. Our data showed that methylation stress impairs E. coli adhesion properties and suggest a possible role of NanA in biofilm formation and bacteria host interactions.