title: Crystal structure of a phosphatase-resistant mutant of sporulation response regulator Spo0F from Bacillus subtilis.
abstract: BACKGROUND
  Spo0F, a phosphotransferase containing an aspartyl pocket, is involved in the signaling pathway (phosphorelay) controlling sporulation in Bacillus subtilis. It belongs to the superfamily of bacterial response regulatory proteins, which are activated upon phosphorylation of an invariant aspartate residue. This phosphorylation is carried out in a divalent cation dependent reaction catalyzed by cognate histidine kinases. Knowledge of the Spo0F structure would provide valuable information that would enable the elucidation of its function as a secondary messenger in a system in which a phosphate is donated from Spo0F to Spo0B, the third of four main proteins that constitute the phosphorelay.
  
  
  RESULTS
  We have determined the crystal structure of a Rap phosphatase resistant mutant, Spo0F Tyr13-->Ser, at 1.9 A resolution. The structure was solved by single isomorphous replacement and anomalous scattering techniques. The overall structural fold is (beta/alpha)5 and contains a central beta sheet. The active site of the molecule is formed by three aspartate residues and a lysine residue which come together at the C terminus of the beta sheet. The active site accommodates a calcium ion.
  
  
  CONCLUSIONS
  The structural analysis reveals that the overall topology and metal-binding coordination at the active site are similar to those of the bacterial chemotaxis response regulator CheY. Structural differences between Spo0F and CheY in the vicinity of the active site provide an insight into how similar molecular scaffolds can be adapted to perform different biological roles by the alteration of only a few amino acid residues. These differences may contribute to the observed stability of the phosphorylated species of Spo0F, a feature demanded by its role as a secondary messenger within the phosphorelay system which controls sporulation.
authors.affiliation: Department of Molecular and Experimental Medicine, Scripps Research Institute, La Jolla, CA 92037, USA.
authors.lastname: Madhusudan
authors.forename: J
authors.initials: J
authors.lastname: Zapf
authors.forename: J M
authors.initials: JM
authors.lastname: Whiteley
authors.forename: J A
authors.initials: JA
authors.lastname: Hoch
authors.forename: N H
authors.initials: NH
authors.lastname: Xuong
authors.forename: K I
authors.initials: KI
authors.lastname: Varughese
mesh_terms: D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000737:chemistry; D001426:Bacterial Proteins / Q000737:chemistry*; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D018360:Crystallography, X-Ray; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000737:chemistry*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012486:Salmonella typhimurium / Q000737:chemistry; D016415:Sequence Alignment; D015398:Signal Transduction; D013171:Spores, Bacterial / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; C080951:Spo0F protein, Bacillus subtilis; D010770:Phosphotransferases; D010744:Phosphoric Monoester Hydrolases; D002118:Calcium