Journal: The Journal of Biological Chemistry
Article Title: Temperature downshifts induce biofilm formation in Pseudomonas aeruginosa through the SiaABCD signal and functional module
doi: 10.1016/j.jbc.2025.111086
Figure Lengend Snippet: The DGC SiaD is primarily responsible for the elevated c-di-GMP content and enhanced Psl production at RT in ATCC 27853. A , relative maximal biofilm biomass of 28 isogenic mutants relative to the ATCC 27853 WT at RT. Blue , genes identified in RNA-seq analysis. B , relative maximal biofilm formation of ATCC 27853 and its isogenic Δ siaD strain at RT and 37 °C. C , relative maximal biofilm formation of ATCC 27853, isogenic Δ siaD mutant, and Δ siaD mutant complemented with rhamnose-inducible expression of siaD at 48 h at RT. VC, the pJM253 vector control. D – F , relative activity of the chromosomal P psl - lux transcriptional fusion reporter ( D ), Psl level in biofilm matrix ( E ), and intracellular c-di-GMP level ( F ) in ATCC 27853 and its isogenic Δ siaD mutant at RT. Midlog phase static cells harvested at 24 h at RT were collected for the analysis in ( C–F ). Data are represented as mean ± SD. (A–E), n = 3 independent experiments. ( F ), n = 2 independent experiments. Asterisks indicate statistical significance. ∗, p < 0.05. ∗∗∗, p < 0.01. ns, not significant (based on Student’s t test). c-di-GMP, cyclic di-guanosine 3′,5′-monophosphate; DGC, diguanylate cyclase.
Article Snippet: LC-MS/MS revealed a reduction in c-di-GMP levels in ATCC 27853 Δ siaD cells compared to the WT ( F ).
Techniques: RNA Sequencing, Mutagenesis, Expressing, Plasmid Preparation, Control, Activity Assay