Bolaturbo & Bola Turbo

 

A turbocharger is a device that takes a low-pressure stream of air from the engine and increases its speed by spinning a shaft inside it. This shaft is connected to a turbine section, which then converts this rotational force into electricity and heat, driving an engine. This process is the main mechanism by which a turbo works, although it is possible for other methods to generate the same result, such as the compression of an explosion. The turbine section is often called the exhaust side of the engine.

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At the transcriptional level, our results show that BolA directly regulates genes involved in the flagellar structure by targeting DNA sequences encoding proteins regulating the flagellar master regulator FlhDC as well as genes encoding proteins that compose the flagellar basal body (fliA and fliZ) or that form the hook-filament junction (flgC, fliF, flgD, and flgJ). In addition, we observed that bolA is able to induce genes related to central carbon metabolism (including transcription of several TCA cycle enzymes), thereby indirectly modulating gene expression required for biofilm formation mechanisms via peptidoglycan synthesis.

We also observed a significant increase in the production of extracellular DNA and proteins, which are crucial components of bacterial adhesion to surfaces, upon bolA overexpression. These changes correlate with the transcriptomic activation of genes encoding proteins that catalyze purine and pyrimidine biosynthesis as well as those involved in the formation of the extracellular matrix — such as the fibronectin-like protein fliA, the collagen-like protein flgC, the chitin synthase flgD, and the cellulose-degrading enzyme flgJ.

In contrast to the downregulation of Bolaturbo genes associated with the flagellar structure, ChIP-seq data show a direct upregulation by BolA of several genes encoding fimbria-like adhesion proteins, such as ydeS, yraH, ygiL, yadC, yadN, yfcV, and yehC. These findings suggest that BolA is a global regulator of the cell, capable of coordinating flagellar and curli synthesis with other cellular pathways required for adhesion to surfaces.

To validate our ChIP-seq results, we performed chromatin immunoprecipitation with antibodies against the 3xFlag tag and then compared the binding profile to that of an isogenic wild-type strain harboring a chromosomal fusion of the bolA gene to a luciferase reporter. Statistically significant peaks were identified, and a consensus sequence was derived by comparison to the sequence of the pool of cloned DNA fragments. The binding sites of BolA are scattered across the chromosome, not only in the promoter regions but also within open reading frames. The graphical representation of the data illustrates that the binding profile of BolA is largely similar to that of the wild-type 3xFlag strain.