title: Cooperativity mutants of the gamma delta resolvase identify an essential interdimer interaction.
abstract: gamma delta resolvase, a transposon-encoded site-specific recombinase, catalyzes the resolution of the cointegrate intermediate of gamma delta transposition. The recombination reaction involves the formation of a catalytic nucleoprotein complex whose structure is determined by specific protein-DNA and protein-protein interactions. We have isolated many resolvase mutants and have identified four that are unable to mediate a subclass of higher order protein-protein interactions necessary for recombination. This mutant phenotype is characterized by an inability to catalyze recombination, a loss of cooperative binding to res DNA, and a failure to induce looping out of the DNA between two resolvase binding sites within res. The amino acid side chains identified by the cooperativity mutants cluster on a surface of the protein that mediates an interaction between resolvase dimers in a crystallographic tetramer. We have therefore identified a region of resolvase that mediates an interdimer protein-protein interaction necessary for the formation of the recombinogenic synaptic intermediate.
authors.affiliation: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06510.
authors.forename: R E
authors.initials: RE
authors.lastname: Hughes
authors.forename: G F
authors.initials: GF
authors.lastname: Hatfull
authors.forename: P
authors.initials: P
authors.lastname: Rice
authors.forename: T A
authors.initials: TA
authors.lastname: Steitz
authors.forename: N D
authors.initials: ND
authors.lastname: Grindley
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004251:DNA Transposable Elements; D046911:Macromolecular Substances; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009713:Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids; D011487:Protein Conformation; D011995:Recombination, Genetic; D019895:Transposases
chemical_list: D004251:DNA Transposable Elements; D046911:Macromolecular Substances; D009713:Nucleotidyltransferases; D019895:Transposases