OmpR and RcsB abolish temporal and spatial changes in expression of flhD in Escherichia coli biofilm.
OmpR and RcsB abolish temporal and spatial changes in expression of flhD in Escherichia coli biofilm.
复制标题
DOI：
10.1186/1471-2180-13-182
复制
发表时间：
2013-08-02
期刊：
影响因子：
4.2
通讯作者：
Prüß BM
中科院分区：
文献类型：
--
作者：
Samanta P;Clark ER;Knutson K;Horne SM;Prüß BM
生物膜是以特定表型为特征的细菌群落，包括对抗微生物剂和宿主免疫系统的抗性增加。这就需要开发新的生物膜预防和治疗方案来对抗传染病。在大肠杆菌中，许多全局调节因子参与了生物膜相关细胞表面细胞器的控制。这些包括鞭毛调节因子FlhD/FlhC、鞭毛调节因子EnvZ/OmpR和可拉酸激活因子RcsCDB。利用流式细胞技术和荧光显微镜技术，我们确定了flhD：：gfp，ompR：：gfp和rcsB：：gfp在E.大肠杆菌生物膜，以及OmpR和RcsB对flhD负调控的影响。从flhD：：gfp研究空间基因表达。flhD的时间基因表达谱在12 h时产生早期峰值，在35 h时表达最小，并且在生物膜发育至51 h时表达第二次增加。相反，ompR曲线在35 h时显示出峰值。ompR的突变在12小时的初始生长期后消除了flhD表达的时间依赖性。有趣的是，rcsB表达与flhD表达不呈负相关，但rcsB的突变也消除了flhD表达的时间依赖性。在空间上，flhD的表达在亲本菌株中生物膜的最外层中最高。在ompR和rcsB突变体中，flhD在整个生物膜中表达。在所有时间和空间实验中，ompR和rcsB中的突变增加flhD表达。在四个测试时间点，生物膜量的减少抵消了这种增加。我们的数据得出结论，FlhD/FlhC及其通过OmpR和RcsB的调节可能是我们开发新型生物膜预防和治疗技术的第一个目标机制。
Biofilms are communities of bacteria that are characterized by specific phenotypes, including an increased resistance towards anti-microbials and the host immune system. This calls for the development of novel biofilm prevention and treatment options to combat infectious disease. In Escherichia coli, numerous global regulators have been implicated in the control of biofilm associated cell surface organelles. These include the flagellar regulator FlhD/FlhC, the osmoregulator EnvZ/OmpR, and the colanic acid activator RcsCDB. Using flow cell technology and fluorescence microscopy, we determined the temporal expression from flhD::gfp, ompR::gfp, and rcsB::gfp in E. coli biofilm, as well as the impact of the negative regulation of flhD by OmpR and RcsB. Spatial gene expression was investigated from flhD::gfp. The temporal gene expression profile for flhD yielded an early peak at 12 h, a minimum of expression at 35 h, and a second increase in expression towards 51 h of biofilm development. In contrast, the ompR profile showed a peak at 35 h. A mutation in ompR abolished time dependence of flhD expression after the initial growth period of 12 h. Intriguingly, rcsB expression did not correlate inversely with flhD expression, yet a mutation in rcsB abolished time dependence of flhD expression as well. Spatially, expression of flhD was highest in the outermost layer of the biofilm in the parent strain. In ompR and rcsB mutants, flhD was expressed throughout the biofilm. Mutations in both, ompR and rcsB increased flhD expression throughout all temporal and spatial experiments. This increase was paralleled by reductions in biofilm amounts at four tested time points. Our data lead to the conclusion that FlhD/FlhC and its regulation by OmpR and RcsB may be our first target mechanism for the development of novel biofilm prevention and treatment techniques.
登录
查看更多内容
影响因子：
3.2
作者：
Kim, DJ;Boylan, B;Forst, S
通讯作者：
Forst, S
影响因子：
5.6
作者：
Ko, M;Park, C
通讯作者：
Park, C
影响因子：
3.2
作者：
Barker, CS;Prüss, BM;Matsumura, P
通讯作者：
Matsumura, P
影响因子：
3.2
作者：
GOTTESMAN, S;TRISLER, P;TORRESCABASSA, A
通讯作者：
TORRESCABASSA, A
影响因子：
64.5
作者：
Boehm, Alex;Kaiser, Matthias;Jenal, Urs
通讯作者：
Jenal, Urs