Novel genetic tools to tackle c-di-GMP-dependent signalling in Pseudomonas aeruginosa.
OBJECTIVE
To develop new genetic tools for studying 3',5'-cyclic diguanylic acid (c-di-GMP) signalling in Pseudomonas aeruginosa.


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.


CONCLUSIONS
The genetic tools developed and validated in this study could facilitate investigations tackling the c-di-GMP signalling process on different fields, from cellular physiology to drug-discovery research.
Department of Science, University Roma Tre, Rome, Italy.
S Vishnu
SV
Pawar
Department of Science, University Roma Tre, Rome, Italy.
M
M
Messina
Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Rome, Italy.
S
S
Rinaldo
Istituto Pasteur-Fondazione Cenci Bolognetti, Department of Biochemical Sciences, Sapienza University of Rome, Rome, Italy.
F
F
Cutruzzolà
Research Core Unit Metabolomics, Institute of Pharmacology, Hannover Medical School, Hannover, Germany.
V
V
Kaever
Department of Science, University Roma Tre, Rome, Italy.
G
G
Rampioni
Department of Science, University Roma Tre, Rome, Italy.
L
L
Leoni
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D005821:Genetic Techniques; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction
D001426:Bacterial Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D006152:Cyclic GMP
Pseudomonas aeruginosa; biofilm inhibition; cyclic-di-GMP; genetic tools; screening; whole-cell biosensors