Evidence for a functional similarity between the two-component regulatory systems RegSR, ActSR, and RegBA (PrrBA) in α-Proteobacteria

Abstract.

The symbiotic bacteria Bradyrhizobium japonicum and Sinorhizobium meliloti, and the purple photosynthetic bacteria Rhodobacter capsulatus, Rhodovulum sulfidophilum, Roseobacter denitrificans and Rhodobacter sphaeroides possess homologous two-component regulatory systems, namely RegSR, ActSR, RegBA and PrrBA. The respective response regulators of these bacteria control expression of different regulons that are involved in N₂ fixation, CO₂ fixation, photosynthesis or acid tolerance. We therefore asked whether the regulators are functionally exchangeable or whether they have disparate functions in the different species, despite the amino acid sequence similarity. In this study, we showed that purified B. japonicum RegR bound in vitro to genuine DNA targets for Rba. capsulatus RegA, and that RegA was phosphorylated in vitro when RegS_C (a soluble variant of the sensor kinase RegS) was added to an Escherichia coli extract containing overexpressed RegA. In vivo, RegA and S. meliloti ActR activated transcription of the B. japonicum fixR-nifA operon, normally a target for RegR. The genes for both regulators, regA and actR, were able to complement a B. japonicum regR mutant with respect to the formation of a nitrogen-fixing symbiosis with soybean. Vice versa, RegR activated in Rba. capsulatus the expression of the photosynthesis operon puc, normally a target for RegA. In conclusion, the results show that B. japonicum RegR, Rba. capsulatus RegA, and S. meliloti ActR are functionally similar.