Bolaturbo & Bola Turbo

 

Bolaturbo & bola turbo

In the current study we investigated the effect of bolA on the regulation of genes related to flagellar biosynthesis and metabolism, using microarrays and ChIP-seq. The results clearly demonstrate that bolA is able to repress the expression of genes involved in a number of key steps in flagellar biosynthesis, as well as modulating additional metabolic pathways connecting to peptidoglycan synthesis, the major component of bacterial cell walls (see Fig. S3 in the supplemental material). In addition, bolA strongly affects the production of polysaccharides such as glycolipids and chitosan, which are involved in the formation of the extracellular matrix in biofilms. The chitosan production was significantly increased in the bolA+ strain, while the glycolipids were significantly repressed.

To elucidate the mode of action of this transcription factor, we performed in silico analysis of the bolA promoter sequence and identified three distinct DNA binding motifs. Using these motifs, we were able to construct a consensus DNA binding site for bolA. This was verified in a number of cloned PCR products using different primers and in all cases the bolA coding region was bound to the consensus DNA fragment.

At the mRNA level, bolA directly regulates 33% of the genes related to flagellar biosynthesis. The repression observed at 1 h and the reduction in motility capacity that occurs 3 h after bolA induction are correlated with these repressions and confirm the direct role of bolA in the regulation of flagellar gene expression. ChIP-seq results also show that bolA is able to directly interact with the mreB promoter and significantly affect its expression.

The overall effect of bolA on central Bola turbo carbon metabolism is demonstrated by the induction of genes encoding amino acid pathways and the significant increase in the transcription of TCA cycle enzymes. In addition, the bolA+ strain exhibits a pronounced resistance to various antibiotics and detergents, confirming that bolA has a strong effect on the ability of bacteria to withstand environmental stressors.

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