The GacS/A-Rsm Pathway Positively Regulates Motility and Flagella Synthesis in Azotobacter vinelandii.

Azotobacter vinelandii is a motile bacterium that possesses an unusual pattern of peritrichous flagellation for members of the Pseudomonadaceae family. Unlike what has been reported for Pseudomonas spp. FleQ is not the master regulator of motility in A. vinelandii, this role is performed by FlhDC. Other factors involved in the regulation of motility are AlgU (σE) and CydR which act as negative regulators. In some members of the Enterobacteriaceae and Pseudomonadaceae families, the GacS/A-Rsm pathway is another important factor regulating motility. In the present study, the involvement of the GacS/A-Rsm pathway in regulating the motility of A. vinelandii was explored; we found that contrary to what has been reported for most of the strains studied of Pseudomonas species, GacS/A, through the Rsm system, positively controlled swimming motility. We show that the target of this regulation is the synthesis of flagella, which most likely occurs in an FlhDC-independent manner.

Study of the sRNA RsmY involved in the genetic regulation of the synthesis of alginate and alkyl resorcinols in Azotobacter vinelandii.

Azotobacter vineladii is a Gram-negative bacterium that produces alginate and poly-hydroxybutyrate (PHB), two polymers of biotechnological interest. This bacterium has the ability to form desiccation-resistant cysts. In the cyst the membrane phospholipids are replaced with a family of phenolic lipids called alkylresorcinols (ARs). The alginate, PHB, and ARs are controlled by the GacS/A two-component system and the small regulatory RNA (sRNA) RsmZ1, belonging to the Rsm (Csr) regulatory system. The Rsm (Csr) systems usually possess two or more sRNAs, in this regard A. vinelandii is the bacterium with the highest number of rsm-sRNAs. Originally, the presence of two sRNAs of the RsmY family (RsmY1 and RsmY2) was reported, but in a subsequent work it was suggested that they conformed to a single sRNA. In this work we provide genetic evidence confirming that rsmY1 and rsmY2 constitute a single gene. Also, it was established that rsmY mutation decreased alginate and ARs production, but did not affect the PHB synthesis. Transcriptional studies showed that rsmY has its higher expression during the stationary growth phase, and in the absence of RsmZ1, rsmY increases its transcription. Interestingly, rsmY expression was influenced by the carbon source, but its expression did not correlate with alginate production.