The RecB subunit of the Escherichia coli RecBCD enzyme couples ATP hydrolysis to DNA unwinding.
The RecB subunit of the Escherichia coli RecBCD enzyme has previously been reported to possess DNA-dependent ATPase activity (Hickson, I. D., Robson, C. N., Atkinson, K. E., Hutton, L., and Emmerson, P. T. (1985) J. Biol. Chem. 260, 1224-1229). Here we demonstrate that a specific interaction between RecB protein and ATP can also be shown by photoaffinity labeling with the ATP analogue 8-azido-ATP. Furthermore, the capacity of the RecB protein to support ATP hydrolysis varies with the structure and length of the DNA cofactor. Single-stranded linear and circular DNA are markedly better in promoting ATP hydrolysis than duplex DNA. The purified RecB protein can function as a DNA helicase, displacing oligonucleotides annealed to viral M13 DNA in an ATP-dependent and orientation-specific manner.
Boehmer
P E
PE
Department of Biochemistry and Genetics, University of Newcastle upon Tyne, Medical School, United Kingdom.
Emmerson
P T
PT
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism*; D000345:Affinity Labels; D001386:Azides / Q000737:chemistry; D001483:Base Sequence; D002384:Catalysis; D002850:Chromatography, Gel; D004247:DNA / Q000378:metabolism*; D004265:DNA Helicases / Q000378:metabolism; D004264:DNA Topoisomerases, Type I / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D006868:Hydrolysis; D007700:Kinetics; D008969:Molecular Sequence Data; D010777:Photochemistry
D000345:Affinity Labels; D001386:Azides; D029968:Escherichia coli Proteins; C017040:8-azidoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D004247:DNA; D005090:Exodeoxyribonucleases; D043211:Exodeoxyribonuclease V; C468322:exodeoxyribonuclease V, E coli; D000251:Adenosine Triphosphatases; D004265:DNA Helicases; D004264:DNA Topoisomerases, Type I