RECOGNITION OF CORE BINDING-SITES BY BACTERIOPHAGE INTEGRASES

Bacteriophage integrases promote recombination between DNA molecules t hat carry attachment sites. They are members of a large and widely dis tributed family of site-specific recombinases with diverse biological roles. The integrases of phages lambda and HK022 are closely related m embers' of this family, but neither protein efficiently recombines the attachment sites of the other phage. The nucleotides responsible for this specificity difference are located close to the points of recombi national strand exchange, within an integrase binding motif called the extended core binding site. There are four imperfectly repeated copie s of this motif in each set of phage attachment sites, but only two, B ' and C, contain major specificity determinants. When these specificit y determinants were replaced by the corresponding nucleotides from a s ite with the alternative specificity, the resulting mutant was recombi ned by both integrases. Thus, the determinants act by impeding recombi nation promoted by the non-cognate integrase. We found that identical nucleotide substitutions within different core site copies had differe nt effects on recombination, suggesting that integrase does not recogn ize each of the extended core binding sites in the same way. Finally, substitution at several positions in lambda integrase with the corresp onding HK022-specific amino acids prevents recombination of lambda att achment sites, and this defect can be suppressed in an allele-specific manner by appropriate substitutions of HK022-specific nucleotides in the extended core binding sites. (C) 1998 Academic Press Limited.