Resolution of Holliday junctions in genetic recombination: RuvC protein nicks DNA at the point of strand exchange.
The RuvC protein of Escherichia coli catalyzes the resolution of recombination intermediates during genetic recombination and the recombinational repair of damaged DNA. Resolution involves specific recognition of the Holliday structure to form a complex that exhibits twofold symmetry with the DNA in an open configuration. Cleavage occurs when strands of like polarity are nicked at the sequence 5'-WTT decreases S-3' (where W is A or T and S is G or C). To determine whether the cleavage site needs to be located at, or close to, the point at which DNA strands exchange partners, Holliday structures were constructed with the junction points at defined sites within this sequence. We found that the efficiency of resolution was optimal when the cleavage site was coincident with the position of DNA strand exchange. In these studies, junction targeting was achieved by incorporating uncharged methyl phosphonates into the DNA backbone, providing further evidence for the importance of charge-charge repulsions in determining DNA structure.
Imperial Cancer Research Fund, Clare Hall Laboratories, South Mimms, Herts, United Kingdom.
R J
RJ
Bennett
S C
SC
West
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D004249:DNA Damage; D003850:Deoxyribonuclease I / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D029968:Escherichia coli Proteins; D007536:Isomerism; D008957:Models, Genetic; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009943:Organophosphorus Compounds / Q000378:metabolism; D011995:Recombination, Genetic
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D009943:Organophosphorus Compounds; C069806:ruvC protein, E coli; C032627:methylphosphonic acid; D004706:Endodeoxyribonucleases; D003850:Deoxyribonuclease I