Repression of ctrA and chpT by a transcriptional regulator of the Xre family that is expressed by RpoN3 and its cognate activator protein in Cereibacter sphaeroides

Cereibacter sphaeroides is an α-proteobacteria that has two flagellar systems. Fla1 directs the assembly of a single subpolar flagellum, and Fla2 directs the assembly of multiple polar flagella. The fla2 genes are controlled by the two-component system CckA/ChpT/CtrA. In the wild-type strain, the fla2 genes are not expressed under the growth conditions commonly used in the laboratory, and thus far, their expression has only been reported in strains carrying either a gain-of-function version of CckA or a null mutation in osp, a negative regulator of CckA. In this work, the differential swimming response of two Fla2 + strains in response to the inclusion of a divalent ion in the culture medium was investigated. This analysis led to identifying a new transcriptional regulator of the XRE family, XrpA. This protein severely reduces the expression of ctrA and, consequently, the expression of the genes activated by this transcription factor. We show that XrpA binds to the control region of ctrA and chpT, suggesting that XrpA directly represses their expression. Additionally, we determined that RpoN3, one of the four RpoN paralogues of RpoN present in C. sphaeroides, and its cognate activator protein AprX are required for the expression of xrpA. XrpA is conserved in several species of Rhodobacterales and a σ54 promoter consensus sequence is present in its control region and a homologue of AprX cooccurs with it. These results support the idea that these proteins form a novel regulatory module that controls the TCS CckA/ChpT/CtrA in C. sphaeroides and other related species.