T7 endonuclease I resolves Holliday junctions formed in vitro by RecA protein.
T7 endonuclease I is known to bind and cleave four-way junctions in DNA. Since these junctions serve as analogues of Holliday junctions that arise during genetic recombination, we have investigated the action of T7 endonuclease I on recombination intermediates containing Holliday junctions. We find that addition of T7 endonuclease I to strand exchange reactions catalysed by RecA protein of Escherichia coli leads to the formation of duplex products that correspond to 'patch' and 'splice' type recombinants. Resolution of the recombination intermediates occurs by the introduction of nicks at the site of the Holliday junction. The recombinant molecules contain 5'-phosphate and 3'-hydroxyl termini which may be ligated to restore the integrity of the DNA.
Müller
B
B
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, UK.
Jones
C
C
West
S C
SC
D004269:DNA, Bacterial / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics*; D011938:Rec A Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic; D013604:T-Phages / Q000201:enzymology*
D004269:DNA, Bacterial; D011938:Rec A Recombinases; D003850:Deoxyribonuclease I