The GGDEF-EAL protein CdgB from Azospirillum baldaniorum Sp245, is a dual function enzyme with potential polar localization

• Published in: PloS one 2022-11, Vol.17 (11), p.e0278036-e0278036
• Main Authors: Viruega-Góngora, Víctor I, Acatitla-Jácome, Iris S, Zamorano-Sánchez, David, Reyes-Carmona, Sandra R, Xiqui-Vázquez, María L, Baca, Beatriz Eugenia, Ramírez-Mata, Alberto
• Format: Article
• Language: English
• Subjects: Analysis; Azo compounds; Azospirillum; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biofilms; Biology and Life Sciences; Cell membranes; Cloning; Cyclic GMP - metabolism; Domains; E coli; Enzymes; Extracellular polymers; Gene Expression Regulation, Bacterial; Genes; Localization; Medicine and Health Sciences; Motility; Mutation; Phosphodiesterase; Phosphoric Diester Hydrolases - metabolism; Plant growth; Plasmids; Properties; Proteins; Research and Analysis Methods; Servers; Swimming
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0278036

Azospirillum baldaniorum Sp245, a plant growth-promoting rhizobacterium, can form biofilms through a process controlled by the second messenger cyclic diguanylate monophosphate (c-di-GMP). A. baldaniorum has a variety of proteins potentially involved in controlling the turnover of c-di-GMP many of which are coupled to sensory domains that could be involved in establishing a mutualistic relationship with the host. Here, we present in silico analysis and experimental characterization of the function of CdgB (AZOBR_p410089), a predicted MHYT-PAS-GGDEF-EAL multidomain protein from A. baldaniorum Sp245. When overproduced, CdgB behaves predominantly as a c-di-GMP phosphodiesterase (PDE) in A. baldaniorum Sp245. It inhibits biofilm formation and extracellular polymeric substances production and promotes swimming motility. However, a CdgB variant with a degenerate PDE domain behaves as diguanylate cyclase (DGC). This strongly suggest that CdgB is capable of dual activity. Variants with alterations in the DGC domain and the MHYT domain negatively affects extracellular polymeric substances production and induction of swimming motility. Surprisingly, we observed that overproduction of CdgB results in increased c-di-GMP accumulation in the heterologous host Escherichia coli, suggesting under certain conditions, the WT CdgB variant can behave predominantly as a DGC. Furthermore, we also demonstrated that CdgB is anchored to the cell membrane and localizes potentially to the cell poles. This localization is dependent on the presence of the MHYT domain. In summary, our results suggest that CdgB can provide versatility to signaling modules that control motile and sessile lifestyles in response to key environmental signals in A. baldaniorum.