title: Step-arrest mutants of FLP recombinase: implications for the catalytic mechanism of DNA recombination.
abstract: The site-specific recombinase (FLP) encoded by the yeast plasmid 2 micron circle belongs to the integrase (of phage lambda) family of recombinases. The sparse homology within the members of this family contrasts with the invariance of three residues, His-396, Arg-399, and Tyr-433 (the numbers correspond to the family alignment positions), among them. We report here results on substrate recognition and catalysis by FLP proteins altered at these residues. Mutations of the conserved His and Tyr that aborted the reaction at specific steps of catalysis permitted genetic dissection of the possible biochemical steps of recombination. We provide indirect evidence that recombination by FLP proceeds through a Holliday junction intermediate.
authors.affiliation: Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, California 92037.
authors.forename: R L
authors.initials: RL
authors.lastname: Parsons
authors.forename: P V
authors.initials: PV
authors.lastname: Prasad
authors.forename: R M
authors.initials: RM
authors.lastname: Harshey
authors.forename: M
authors.initials: M
authors.lastname: Jayaram
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D010582:Bacteriophage lambda / Q000235:genetics; D001483:Base Sequence; D004254:DNA Nucleotidyltransferases / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011995:Recombination, Genetic; D013379:Substrate Specificity
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase