Insights into signal transduction revealed by the low resolution structure of the FixJ response regulator.
Two-component regulatory systems mediate most of the bacterial cells responses to a variety of signals. In Sinorhizobium meliloti, the FixL-FixJ couple controls the expression of the nitrogen fixation genes through the binding of the two-domains response regulator FixJ to the fixK and nifA promoters. Phosphorylation of the N-terminal regulatory domain activates the protein and releases the inhibition of the C-terminal DNA-binding domain that occurs in the unphosphorylated protein. Insights into the transition from the inactive to the active form are provided by the architecture of the unphosphorylated response regulator reported in this study. The relative position and orientation of the N and C-terminal domains were defined from the molecular envelope restored from small-angle X-ray scattering (SAXS) data. The involvement of the alpha4-beta5-alpha5 surface of the regulatory domain, the linker region and the C-terminal helix of the DNA-binding domain in the interdomain interface of unphosphorylated FixJ was supported by biochemical investigations. These results, together with the previously reported studies on the phosphorylated regulatory domain of FixJ, emphasize the role of the alpha4-beta5-alpha5 surface in mediating a flow of information in this response regulator. This first study by SAXS of proteins from two-component systems suggests that the method could be successfully applied to other members of this family and could be suitable for the study of multidomain proteins and protein-protein complexes regulated through molecular interfaces in the low micromolar range.
Birck
Catherine
C
Groupe de Cristallographie Biologique, CNRS-IPBS, 205 route de Narbonne, 31077- Cedex, Toulouse, France.
Malfois
Marc
M
Svergun
Dmitri
D
Samama
Jean
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009586:Nitrogen Fixation / Q000235:genetics; D010766:Phosphorylation; D011487:Protein Conformation; D012542:Scattering, Radiation; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D016962:Sinorhizobium meliloti / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; C069289:FixJ protein, Bacteria