Analysis of the BadR regulon in Borrelia burgdorferi

Borrelia burgdorferi, the causative agent of Lyme disease, relies on tightly coordinated gene expression to quickly adapt and survive in the tick vector and mammalian host. BadR, an ROK (repressor, open reading frame, kinase) family transcriptional regulator, binds directly to B. burgdorferi promoter DNA, however, many questions concerning the role for BadR in gene regulation remain unanswered. In particular, there are conflicting reports concerning what genes are regulated by BadR in B. burgdorferi. Furthermore, previous studies have suggested important roles for BadR in unfed ticks, but the BadR regulon has not been defined under such conditions. Additionally, although BadR regulates rpoS expression in a growth phase-dependent manner, it remains unknown whether BadR regulates other genes during different growth phases.

To address these questions, we cultivated a B. burgdorferi badR mutant and wild-type strain under various conditions and analyzed the transcriptome using RNA-sequencing. When spirochetes were grown at 37 °C and collected at the mid-logarithmic and stationary phase of growth, 211 and 272 genes were differentially expressed in the badR mutant, respectively. A total of 79 genes were differentially expressed when spirochetes were grown at 23 °C. A vast majority of genes identified in this study encode proteins of unknown function.

Complex transcriptional regulation mechanisms coordinate the expression of genes required for the survival of B. burgdorferi throughout its tick-mammal enzootic lifecycle. As part of this process, BadR functions as a global regulatory protein and regulates B. burgdorferi virulence gene expression. Combined, this work supports a role for BadR in global B. burgdorferi gene regulation by modulating expression of different sets of genes at different stages of the enzootic lifecycle. We anticipate that investigating the function of genes in the BadR regulon will lead to the identification of novel virulence factors for therapeutic and vaccine development.