Cooperativity mutants of the gamma delta resolvase identify an essential interdimer interaction.
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.
Hughes
R E
RE
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06510.
Hatfull
G F
GF
Rice
P
P
Steitz
T A
TA
Grindley
N D
ND
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
D004251:DNA Transposable Elements; D046911:Macromolecular Substances; D009713:Nucleotidyltransferases; D019895:Transposases