OmpA influences Escherichia coli biofilm formation by repressing cellulose production through the CpxRA two-component system
OmpA influences Escherichia coli biofilm formation by repressing cellulose production through the CpxRA two-component system
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DOI：
10.1111/j.1462-2920.2009.02000.x
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发表时间：
2009-10-01
影响因子：
5.1
通讯作者：
Wood, Thomas K.
中科院分区：
文献类型：
--
作者：
Ma, Qun;Wood, Thomas K.
P>之前我们发现大肠杆菌的 OmpA 会增加聚苯乙烯表面生物膜的形成（Gonzalez Barrios 等人，Biotechnol Bioeng，93：188-200，2006a）。在这里，我们表明 OmpA 对疏水性和亲水性表面生物膜形成的影响不同，因为它抑制亲水性纤维素的产生。 OmpA 增加了聚苯乙烯、聚丙烯和聚乙烯表面上的生物膜形成，同时减少了玻璃表面上的生物膜形成。沙柱试验证实 OmpA 减少了对亲水表面的附着。 ompA突变体形成粘性集落，引起粘性的胞外多糖被鉴定为纤维素。全转录组研究表明，OmpA 诱导响应膜应激的 CpxRA 双组分信号转导通路。 CpxA 磷酸化 CpxR 并导致 csgD 表达减少。 CsgD 产量减少会抑制 adrA 表达，导致纤维素产量减少，因为 CsgD 和 AdrA 负责 3,5-环二鸟苷酸和纤维素合成。实时聚合酶链反应证实 csgD 和 adrA 被 OmpA 抑制。使用双缺失突变体 adrA ompA、csgB ompA 和 cpxR ompA 进行的生物膜和纤维素测定证实，OmpA 通过 CpxR 和 AdrA 减少了纤维素产生并增加了聚苯乙烯表面上的生物膜形成。 OmpA 和 CpxRA 系统之间联系的进一步证据是 OmpA 的过量产生会破坏细胞膜并导致细胞裂解。因此，OmpA 通过 CpxRA 应激反应系统抑制纤维素的产生，并且纤维素的减少增加了疏水表面上生物膜的形成。
P>Previously we discovered that OmpA of Escherichia coli increases biofilm formation on polystyrene surfaces (Gonzalez Barrios et al., Biotechnol Bioeng, 93:188-200, 2006a). Here we show OmpA influences biofilm formation differently on hydrophobic and hydrophilic surfaces since it represses cellulose production which is hydrophilic. OmpA increased biofilm formation on polystyrene, polypropylene, and polyvinyl surfaces while it decreased biofilm formation on glass surfaces. Sand column assays corroborated that OmpA decreases attachment to hydrophilic surfaces. The ompA mutant formed sticky colonies, and the extracellular polysaccharide that caused stickiness was identified as cellulose. A whole-transcriptome study revealed that OmpA induces the CpxRA two-component signal transduction pathway that responds to membrane stress. CpxA phosphorylates CpxR and results in reduced csgD expression. Reduced CsgD production represses adrA expression and results in reduced cellulose production since CsgD and AdrA are responsible for 3,5-cyclic diguanylic acid and cellulose synthesis. Real-time polymerase chain reaction confirmed csgD and adrA are repressed by OmpA. Biofilm and cellulose assays with double deletion mutants adrA ompA, csgB ompA, and cpxR ompA confirmed OmpA decreased cellulose production and increased biofilm formation on polystyrene surfaces through CpxR and AdrA. Further evidence of the link between OmpA and the CpxRA system was that overproduction of OmpA disrupted the membrane and led to cell lysis. Therefore, OmpA inhibits cellulose production through the CpxRA stress response system, and this reduction in cellulose increases biofilm formation on hydrophobic surfaces.