Document Type
Article
Publication Date
5-2012
Publication Source
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.
Inclusive pages
5060-5069
ISBN/ISSN
0099-2240
Document Version
Published Version
Copyright
Copyright © 2012, American Society for Microbiology
Publisher
American Society for Microbiology
Volume
78
Peer Reviewed
yes
Issue
15
eCommons Citation
Rybtke, Morten T.; Borlee, Bradley R.; Murakami, Keiji; Irie, Yasuhiko; Hentzer, Morten; Nielsen, Thomas E.; Givskov, Michael; Parsek, Matthew R.; and Tolker-Nielsen, Tim, "Fluorescence-Based Reporter for Gauging Cyclic Di-GMP Levels in Pseudomonas aeruginosa" (2012). Biology Faculty Publications. 207.
https://ecommons.udayton.edu/bio_fac_pub/207
Included in
Biology Commons, Biotechnology Commons, Cell Biology Commons, Genetics Commons, Microbiology Commons, Molecular Genetics Commons
Comments
This document is provided for download in compliance with the publisher's policy on self-archiving. Permission documentation is on file.