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|Title:||Fluorescence-based reporter for gauging cyclic Di-GMP levels in Pseudomonas aeruginosa||Authors:||Borlee, Bradley R.
Nielsen, Thomas E.
Parsek, Matthew R.
Rybtke, Morten Theil
|Issue Date:||2012||Source:||Rybtke, M. T., Borlee, B. R., Murakami, K., Irie, Y., Hentzer, M., Nielsen, T. E., et al. (2012). Fluorescence-based reporter for gauging cyclic Di-GMP levels in Pseudomonas aeruginosa. Applied and Environmental Microbiology, 78(15), 5060-5069.||Series/Report no.:||Applied and environmental microbiology||Abstract:||The increased tolerance toward the host immune system and antibiotics displayed by biofilm-forming Pseudomonas aeruginosa and other bacteria in chronic infections such as cystic fibrosis bronchopneumonia is of major concern. Targeting of biofilm formation is believed to be a key aspect in the development of novel antipathogenic drugs that can augment the effect of classic antibiotics by decreasing antimicrobial tolerance. The second messenger cyclic di-GMP is a positive regulator of biofilm formation, and cyclic di-GMP signaling is now regarded as a potential target for the development of antipathogenic compounds. Here we describe the development of fluorescent monitors that can gauge the cellular level of cyclic di-GMP in P. aeruginosa. We have created cyclic di-GMP level reporters by transcriptionally fusing the cyclic di-GMP-responsive cdrA promoter to genes encoding green fluorescent protein. We show that the reporter constructs give a fluorescent readout of the intracellular level of cyclic di-GMP in P. aeruginosa strains with different levels of cyclic di-GMP. Furthermore, we show that the reporters are able to detect increased turnover of cyclic di-GMP mediated by treatment of P. aeruginosa with the phosphodiesterase inducer nitric oxide. Considering that biofilm formation is a necessity for the subsequent development of a chronic infection and therefore a pathogenicity trait, the reporters display a significant potential for use in the identification of novel antipathogenic compounds targeting cyclic di-GMP signaling, as well as for use in research aiming at understanding the biofilm biology of P. aeruginosa.||URI:||https://hdl.handle.net/10356/100527
|ISSN:||0099-2240||DOI:||http:dx.doi.org/10.1128/AEM.00414-12||Rights:||© 2012 American Society for Microbiology.||Fulltext Permission:||none||Fulltext Availability:||No Fulltext|
|Appears in Collections:||SCELSE Journal Articles|
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