Borrelia burgdorferi rel is responsible for generation of guanosine-3′-diphosphate-5′-triphosphate and growth control

The global transcriptional regulator (p)ppGpp (guanosine-3'-diphosphate-5'-triphosphate and guanosine-3',S'-bisphosphate, collectively) produced by the relA and spoT genes in Escherichia coli allows bacteria to adapt to different environmental stresses. The genome of Borrelia burgdorferi encodes a single chromosomal rel gene (11130198) (B. burgdorferi rei [rel(Bbu)]) homologous to relA and spoT of E. coli. Its role in (p)ppGpp synthesis, bacterial growth, and modulation of gene expression has not been studied in detail. We constructed a rel(Bbu). deletion mutant in an infectious B. burgdorferi 297 strain and isolated an extrachromosomally complemented derivative of this mutant. The mutant did not synthesize rel(Bbu) mRNA, Rel(Bbu) protein, or (p)ppGpp. This synthesis was restored in the complemented derivative, confirming that rel(Bbu). is necessary and sufficient for (p)ppGpp synthesis and degradation in B. burgdorferi. The rel(Bbu), mutant grew well during log phase in complete BSK-H but reached lower cell concentrations in the stationary phase than the wild-type parent, suggesting that (p)ppGpp may be an important factor in the ability of B. burgdorferi to adapt to stationary phase. Deletion of rel(Bbu) did not eliminate the temperature-elicited OspC shift, nor did it alter bmp gene expression or B. burgdorferi antibiotic susceptibility. Although deletion of rel(Bbu) eliminated B. burgdorferi virulence for mice, which was not restored by complementation, we suggest that rel(Bbu).-dependent accumulation of (p)ppGpp may be important for in vivo survival of this pathogen.