Recognition of core binding sites by bacteriophage integrases.
Bacteriophage integrases promote recombination between DNA molecules that carry attachment sites. They are members of a large and widely distributed family of site-specific recombinases with diverse biological roles. The integrases of phages lambda and HK022 are closely related members 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 recombinational strand exchange, within an integrase binding motif called the extended core binding site. There are four imperfectly repeated copies of this motif in each set of phage attachment sites, but only two, B' and C, contain major specificity determinants. When these specificity determinants were replaced by the corresponding nucleotides from a site with the alternative specificity, the resulting mutant was recombined by both integrases. Thus, the determinants act by impeding recombination promoted by the non-cognate integrase. We found that identical nucleotide substitutions within different core site copies had different effects on recombination, suggesting that integrase does not recognize each of the extended core binding sites in the same way. Finally, substitution at several positions in lambda integrase with the corresponding HK022-specific amino acids prevents recombination of lambda attachment 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.
Dorgai
L
L
Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, Bethesda, MD 20892, USA.
Sloan
S
S
Weisberg
R A
RA
D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000201:enzymology*; D001665:Binding Sites / Q000235:genetics; D003090:Coliphages / Q000201:enzymology; D019426:Integrases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic / Q000235:genetics; D016415:Sequence Alignment; D014764:Viral Proteins / Q000378:metabolism
D014764:Viral Proteins; D019426:Integrases