Genetic Dissection of Cyclic di‐GMP Signalling in Pseudomonas aeruginosa via Systematic Diguanylate Cyclase Disruption DOI Creative Commons
Román A. Martino, Daniel C. Volke, Albano H. Tenaglia

et al.

Microbial Biotechnology, Journal Year: 2025, Volume and Issue: 18(4)

Published: April 1, 2025

The second messenger bis-(3' → 5')-cyclic dimeric guanosine monophosphate (c-di-GMP) governs adaptive responses in the opportunistic pathogen Pseudomonas aeruginosa, including biofilm formation and transition from acute to chronic infections. Understanding intricate c-di-GMP signalling network remains challenging due overlapping activities of numerous diguanylate cyclases (DGCs). In this study, we employed a CRISPR-based multiplex genome-editing tool disrupt all 32 GGDEF domain-containing proteins (GCPs) implicated P. aeruginosa PA14. Phenotypic physiological analyses revealed that resulting mutant was unable form biofilms had attenuated virulence. Residual levels were still detected despite extensive GCP disruption, underscoring robustness regulatory network. Taken together, these findings provide insights into complex metabolism showcase importance functional bacterial signalling. Moreover, our approach overcomes native redundancy synthesis, providing framework dissect individual DGC functions paving way for targeted strategies address adaptation pathogenesis.

Language: Английский

Genetic Dissection of Cyclic di‐GMP Signalling in Pseudomonas aeruginosa via Systematic Diguanylate Cyclase Disruption DOI Creative Commons
Román A. Martino, Daniel C. Volke, Albano H. Tenaglia

et al.

Microbial Biotechnology, Journal Year: 2025, Volume and Issue: 18(4)

Published: April 1, 2025

The second messenger bis-(3' → 5')-cyclic dimeric guanosine monophosphate (c-di-GMP) governs adaptive responses in the opportunistic pathogen Pseudomonas aeruginosa, including biofilm formation and transition from acute to chronic infections. Understanding intricate c-di-GMP signalling network remains challenging due overlapping activities of numerous diguanylate cyclases (DGCs). In this study, we employed a CRISPR-based multiplex genome-editing tool disrupt all 32 GGDEF domain-containing proteins (GCPs) implicated P. aeruginosa PA14. Phenotypic physiological analyses revealed that resulting mutant was unable form biofilms had attenuated virulence. Residual levels were still detected despite extensive GCP disruption, underscoring robustness regulatory network. Taken together, these findings provide insights into complex metabolism showcase importance functional bacterial signalling. Moreover, our approach overcomes native redundancy synthesis, providing framework dissect individual DGC functions paving way for targeted strategies address adaptation pathogenesis.

Language: Английский

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