OmpR and RcsB abolish temporal and spatial changes in expression of flhD in Escherichia coli Biofilm

BMC Microbiology

Birgit M Prüß¹  Shelley M Horne¹  Katie Knutson¹  Emily R Clark¹  Priyankar Samanta¹ 

[1] Department of Veterinary and Microbiological Sciences, North Dakota State University, Fargo, ND 58108, USA

关键词: Fluorescence microscopy;    Reporter gene fusion;    Biofilm;    Escherichia coli;   

Others  :  1143280
DOI  :  10.1186/1471-2180-13-182

 received in 2013-04-11, accepted in 2013-07-31,  发布年份 2013

【 摘 要 】

Background

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.

Results

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.

Conclusion

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.

【 授权许可】

   
2013 Samanta et al.; licensee BioMed Central Ltd.

【 图 表 】

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