Structural Basis of a Novel Heme Binding Bacterial One-Component Switch
One-component systems (OCSs) integrate sensory and effector functions within a single protein, enabling rapid gene expression changes in response to environmental cues. Here, we characterized a novel heme binding OCS protein, FG214, from Fimbriimonas ginsengisoli, a redox-regulated helix-turn-helix transcription factor in which heme iron ligand state controls a monomer-to-dimer switch. Data supporting this included our observation of the FG214 PAS domain binding a hexacoordinate heme b in oxidized conditions and undergoing a slate of redox and ligand-dependent conformational changes, transitioning from a monomer to a homodimer. Spectroscopic and structural data revealed that oxidation stabilizes the likely HTH-PAS intramolecular domain interface, while reduction of the heme iron dissociates the HTH, freeing previously-sequestered homodimerization surfaces. Similar effects were seen by addition of a small molecule ferric heme ligand, as directly visualized with a 1.47 [A] crystal structure of an imidazole-bound truncated construct. Using in vitro DNA-binding assays, we identified an artificial promoter sequence and demonstrated ligand-enhanced protein-DNA binding. Finally, we performed in vivo proof of concept experiments establishing FG214 as a redox-sensitive scaffold for biosensor engineering. Together, these findings define FG214 as a novel heme-binding PAS DNA binding protein, complementing known heme-PAS two-component signaling switches.