Kinetic modeling of the recA protein promoted renaturation of complementary DNA strands.
Quantitative agarose gel assays reveal that the recA protein promoted renaturation of complementary DNA strands (phi X DNA) proceeds in two stages. The first stage results in the formation of unit-length duplex DNA as well as a distribution of other products ("initial products"). In the second stage, the initial products are converted to complex multipaired DNA structures ("network DNA"). In the presence of ATP, the initial products are formed within 2 min and are then rapidly converted to network DNA. In the absence of ATP, the initial products are formed nearly as fast as with ATP present, but they are converted to network DNA at a much lower rate. The time-dependent formation of initial products and network DNA from complementary single strands for both the ATP-stimulated and ATP-independent reactions can be modeled by using a simple two-step sequential kinetic scheme. This model indicates that the primary effect of ATP in the recA protein promoted renaturation reaction is not on the initial pairing step (which leads to the formation of initial products) but rather is to increase the rate at which subsequent pairing events can occur.
Bryant
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FR
Department of Biochemistry, Johns Hopkins University, School of Hygiene and Public Health, Baltimore, Maryland 21205.
Menge
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KL
Nguyen
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D000255:Adenosine Triphosphate / Q000378:metabolism; D010584:Bacteriophage phi X 174 / Q000378:metabolism; D004270:DNA, Circular / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism*; D007700:Kinetics; D008962:Models, Theoretical; D009695:Nucleic Acid Renaturation; D011485:Protein Binding; D011938:Rec A Recombinases / Q000378:metabolism*
D004270:DNA, Circular; D004279:DNA, Viral; D000255:Adenosine Triphosphate; D011938:Rec A Recombinases