Cleavage of double-crossover molecules by T4 endonuclease VII.
DNA double-crossover molecules containing two Holliday junctions have been prepared and treated with endonuclease VII, the resolvase from bacteriophage T4. One molecule contains antiparallel double-helical domains, and the other molecule contains parallel domains. The parallel double-crossover model system has been made tractable by closing the free ends of the molecule, to convert it to a catenane. The products resulting from the two substrates differ substantially. The molecule containing antiparallel helical domains is cleaved three nucleotides 3' to the crossover points, in a fashion similar to single Holliday junction analogs. The molecule containing parallel helical domains is cleaved, but the major points of scission are five nucleotides 5' to a branch point on the crossover strands and six nucleotides 3' to the same branch point on the non-crossover strands. The major sites of scission reflect features of molecular symmetry in each case, suggesting that the resolvase recognizes structural features. The cleavage results suggest that the antiparallel structure is the natural substrate, if the Holliday junction is unconstrained within the cell. It is straightforward to reconcile antiparallel Holliday junctions with the conventional parallel paradigm of recombination. Nevertheless, the cleavage of the parallel molecule shows that a parallel substrate could also be cleaved symmetrically by endonuclease VII (but with different products) if the molecule were constrained to assume that conformation within the cell.
Fu
T J
TJ
Institute of Genetics, University of Köln, Germany.
Kemper
B
B
Seeman
N C
NC
D001483:Base Sequence; D004247:DNA / Q000378:metabolism*; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005609:Free Radicals; D006878:Hydroxides / Q000737:chemistry; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011995:Recombination, Genetic; D013379:Substrate Specificity
D005609:Free Radicals; D006878:Hydroxides; D004247:DNA; D004706:Endodeoxyribonucleases; C035571:endodeoxyribonuclease VII