Two forms of RecA-single-stranded DNA-adenosine 5'-O-(3-thiotriphosphate) complexes with different activities for cleavage of phage phi 80 cI repressor.
The kinetics of cleavage of the phage phi 80 cI repressor by Escherichia coli RecA protein were studied. The rate of cleavage in the presence of single-stranded DNA (ssDNA) and either adenosine 5'-O-(3-thiotriphosphate) (ATP gamma S), ATP or dATP is very low in the first hour at 37 degrees C and then increases sharply as incubation continues. The initial rate of cleavage of the repressor is greatly increased by incubating the RecA protein with ssDNA prior to addition of ATP gamma S and the repressor. However, when ATP gamma S is present during preincubation of RecA protein with ssDNA, the stimulatory effect of preincubation is greatly reduced. This difference in the effect of preincubation in two different conditions can be explained by formation of RecA-ssDNA-ATP gamma S complexes with different activities for cleavage of the repressor. The active complex is formed by binding of ATP gamma S to a complex of RecA protein and ssDNA. However, when the RecA protein binds to ATP gamma S prior to its binding to ssDNA, the resulting complex has no or only very weak cleavage activity toward the repressor.
Eguchi
Y
Y
Department of Biology, Faculty of Science, Osaka University, Japan.
Ogawa
T
T
Ogawa
H
H
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D001435:Bacteriophages / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004926:Escherichia coli; D007700:Kinetics; D011938:Rec A Recombinases / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism*; D013997:Time Factors; D014157:Transcription Factors / Q000378:metabolism*; D014364:Tryptophan / Q000235:genetics; D014764:Viral Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D012097:Repressor Proteins; D014157:Transcription Factors; D014764:Viral Proteins; C022571:adenosine 5'-O-(3-thiotriphosphate); D014364:Tryptophan; D000255:Adenosine Triphosphate; D011938:Rec A Recombinases