X-ray structure of aminopeptidase A from Escherichia coli and a model for the nucleoprotein complex in Xer site-specific recombination

The structure of aminopeptidase A (PepA), which functions as a DNA-binding protein in Xer site-specific recombination and in transcriptional control o f the carAB operon in Escherichia coli, has been determined at 2.5 ij resol ution. In Xer recombination at cer, PepA and the arginine repressor (ArgR) serve as accessory proteins, ensuring that recombination is exclusively int ramolecular, In contrast, PepA homologues from other species have no known DNA-binding activity and are not implicated in transcriptional regulation o r control of site-specific recombination, PepA comprises two domains, which have similar folds to the two domains of bovine lens leucine aminopeptidas e (LAP), However, the N-terminal domain of PepA, which probably plays a sig nificant role in DNA binding, is rotated by 19 degrees compared with its po sition in LAP. PepA is a homohexamer of 32 symmetry, A groove that runs fro m one trimer face across the 2-fold molecular axis to the other trimer face is proposed to be the DNA-binding site. Molecular modelling supports a str ucture of the Xer complex in which PepA, ArgR and a second PepA molecule ar e sandwiched along their 3-fold molecular axes, and the accessory sequences of the two recombination sites wrap around the accessory proteins as a rig ht-handed superhelix such that three negative supercoils are trapped.