Analysis of double stranded DNA-dependent activities of Deinococcus radiodurans RecA protein.
In this study, the double-stranded DNA-dependent activities of Deinococcus radiodurans RecA protein (Dr RecA) were characterized. The interactions of the Dr RecA protein with double-stranded DNA were determined, especially dsDNA-dependent ATP hydrolysis by the Dr RecA protein and the DNA strand exchange reaction, in which multiple branch points exist on a single RecA protein-DNA complex. A nucleotide cofactor (ATP or dATP ) was required for the Dr RecA protein binding to duplex DNA. In the presence of dATP, the nucleation step in the binding process occurred more rapidly than in the presence of ATP. Salts inhibited the binding of the Dr RecA protein to double-stranded DNA. Double-stranded DNA-dependent ATPase activities showed a different sensitivity to anion species. Glutamate had only a minimal effect on the double-stranded DNA-dependent ATPase activities, up to a concentration of 0.7 M. In the competition experiment for Dr RecA protein binding, the Dr RecA protein manifested a higher affinity to double-stranded DNA than was observed for single-stranded DNA.
Kim
Jong-Il
JI
Department of Food and Microbial Technology, Seoul Women's University, 126 Kongnung 2-dong, Nowon-Gu, Seoul 139-774, Republic of Korea. jikim@swu.ac.kr
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism*; D034301:Deinococcus / Q000378:metabolism*; D003838:Deoxyadenine Nucleotides / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay / Q000379:methods; D004789:Enzyme Activation / Q000187:drug effects; D018698:Glutamic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D011189:Potassium Chloride / Q000494:pharmacology; D011485:Protein Binding; D011938:Rec A Recombinases / Q000378:metabolism*; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors
D004269:DNA, Bacterial; D003838:Deoxyadenine Nucleotides; D018698:Glutamic Acid; D012965:Sodium Chloride; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D004247:DNA; D011938:Rec A Recombinases; D000251:Adenosine Triphosphatases; C026600:2'-deoxyadenosine triphosphate