Pseudomonas aeruginosa post‐translational responses to elevated c‐di‐GMP levels

Abstract

C‐di‐GMP signaling can directly influence bacterial behavior by affecting the functionality of c‐di‐GMP‐binding proteins. In addition, c‐di‐GMP can exert a global effect on gene transcription or translation, for example, via riboswitches or by binding to transcription factors. In this study, we investigated the effects of changes in intracellular c‐di‐GMP levels on gene expression and protein production in the opportunistic pathogen Pseudomonas aeruginosa. We induced c‐di‐GMP production via an ectopically introduced diguanylate cyclase and recorded the transcriptional, translational as well as proteomic profile of the cells. We demonstrate that rising levels of c‐di‐GMP under growth conditions otherwise characterized by low c‐di‐GMP levels caused a switch to a non‐motile, auto‐aggregative P. aeruginosa phenotype. This phenotypic switch became apparent before any c‐di‐GMP‐dependent role on transcription, translation, or protein abundance was observed. Our results suggest that rising global c‐di‐GMP pools first affects the motility phenotype of P. aeruginosa by altering protein functionality and only then global gene transcription.

Characterization of a soluble library of thePseudomonas aeruginosaPAO1 membrane proteome with emphasis on c-di-GMP turnover enzymes

“…Most of the identified proteins are membrane-integrated or likely to be membrane-associated ( Table 1 ). To our knowledge, the dataset presented in this work is thus far the largest set of proteins detected from the c-di-GMP network of P. aeruginosa (Blonder et al 2004 , Düvel et al 2012 , Casabona et al 2013 , Kumari et al 2014 , Bense et al 2022 ). In our nanodiscs library, GGDEF/EAL proteins were detected together with their described interaction partner(s) in the overlapping fractions, indicating preservation of these interactions.…”

Transcriptome Dynamics of Pseudomonas aeruginosa during Transition from Overlapping To Non-Overlapping Cell Cycles

“…Bacteria were fixed by addition of glutaraldehyde (final concentration 2%) for 30 min, and addition of formaldehyde (final concentration 5%) into the culture medium. EM sample preparation was performed as previously described ( 44 ) with slight modifications. In brief, samples were washed twice with TE-buffer and fixed to poly-l-lysine coated round coverslips.…”

Inhibitory Effects of the Fungal Pigment Rubiginosin C on Hyphal and Biofilm Formation in Candida albicans and Candida auris

“…A 50 µL aliquot was added to round poly-L-lysine pretreated coverslips and incubated (room temperature, 10 min). Further processing was carried out as previously described with slight modifications [45]. In brief, cells were fixed for 10 min on the coverslip with TE buffer including 1% glutaraldehyde (final concentration).…”