title: Isolation and characterization of intermediates in site-specific recombination.
abstract: Cre, the site-specific recombinase from bacteriophage P1, catalyzes a recombination reaction between specific DNA sequences designated as lox sites. The breakage and rejoining of partners during this recombination process must be highly concerted because it has not been possible to detect intermediates of the reaction with wild-type Cre. Several mutant Cre proteins have been isolated that produce significant amounts of a possible intermediate product of the recombination reaction. The product has been identified as a Holliday structure in which one set of the DNA strands of the recombining partners has been exchanged. Wild-type Cre protein is capable of acting on this structure to form recombinant products, which is consistent with this being an intermediate in the recombination reaction. Characterization of the Holliday structure indicated that one set of strands in the recombining partners was always exchanged preferentially before the other set. In addition, it has been found that certain Cre mutants that are unable to carry out recombination in vitro are able to resolve the intermediate. This suggests that these mutants are defective in a step in the reaction that precedes the formation of the Holliday intermediate.
authors.affiliation: Experimental Station, E.I. Dupont de Nemours & Co., Wilmington, DE 19898.
authors.forename: R
authors.initials: R
authors.lastname: Hoess
authors.forename: A
authors.initials: A
authors.lastname: Wierzbicki
authors.forename: K
authors.initials: K
authors.lastname: Abremski
mesh_terms: D003090:Coliphages / Q000235:genetics*; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D004279:DNA, Viral / Q000235:genetics / Q000648:ultrastructure; D004926:Escherichia coli / Q000235:genetics*; D019426:Integrases; D009154:Mutation; D011995:Recombination, Genetic; D014764:Viral Proteins
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins; C045073:Cre recombinase; D004254:DNA Nucleotidyltransferases; D019426:Integrases; D004262:DNA Restriction Enzymes