AIMS: To develop new genetic tools for studying 3',5'-cyclic diguanylic acid (c-di-GMP) signaling in Pseudomonas aeruginosa. METHODS AND RESULTS: Plasmid pPcdrA::lux, carrying a transcriptional fusion between the c-di-GMP responsive promoter PcdrA and the luxCDABE reporter genes, has been generated and validated in purpose-built P. aeruginosa strains in which c-di-GMP levels can be increased or reduced upon arabinose-dependent induction of c-di-GMP synthetizing or degrading enzymes. CONCLUSIONS: The reporter systems described so far were able to detect a decrease in the c-di-GMP levels only in engineered strains overproducing c-di-GMP. Conversely, pPcdrA::lux could be used for studying any process or chemical compound expected to cause both an increase or a decrease with respect to the c-di-GMP levels produced by wild type P. aeruginosa. Another relevant aspect of this study has been the development of novel and improved genetic devices for the fine arabinose-dependent control of c-di-GMP levels in P. aeruginosa. SIGNIFICANCE AND IMPACT OF THE STUDY: The genetic tools developed and validated in this study could facilitate investigations tackling the c-di-GMP signaling process on different fields, from cellular physiology to drug-discovery research. This article is protected by copyright. All rights reserved.
Vishnu Pawar, S., Messina, M., Rinaldo, S., Cutruzzolà, F., Kaever, V., Rampioni, G., et al. (2016). Novel genetic tools to tackle c-di-GMP-dependent signaling in Pseudomonas aeruginosa. JOURNAL OF APPLIED MICROBIOLOGY, 120(1), 205-217 [10.1111/jam.12984].