Functional analysis of Arg-308 mutants of Flp recombinase. Possible role of Arg-308 in coupling substrate binding to catalysis.
The arginine residue at position 308 in the Flp recombinase corresponds to the only invariant arginine within the Int family of recombinases. Alterations of this residue result in Flp variants that retain substrate recognition, but form weaker protein-DNA complexes than wild type Flp. Furthermore, their DNA cleavage activity is significantly diminished. A conservative change of R308K results in a functional Flp variant; however, this protein has a lowered temperature optimum for recombination. The Arg-308 mutants can be stabilized on the DNA substrate through cooperativity with a partner Flp mutant that is tight binding. Thus, interactions between Flp monomers must be a relevant feature of the normal recombination reaction.
Research Institute of Scripps Clinic, La Jolla, California 92037.
R L
RL
Parsons
B R
BR
Evans
L
L
Zheng
M
M
Jayaram
D001120:Arginine; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D002384:Catalysis; D004247:DNA / Q000378:metabolism*; D004254:DNA Nucleotidyltransferases / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D013329:Structure-Activity Relationship; D013696:Temperature
D001425:Bacterial Outer Membrane Proteins; D004247:DNA; D001120:Arginine; D004254:DNA Nucleotidyltransferases; C045119:FLP recombinase