Near-simultaneous DNA cleavage by the subunits of the junction-resolving enzyme T4 endonuclease VII.
In common with a number of other DNA junction-resolving enzymes, endonuclease VII of bacteriophage T4 binds to a four-way DNA junction as a dimer, and cleaves two strands of the junction. We have used a supercoil-stabilized cruciform substrate to probe the simultaneity of cleavage at the two sites. Active endonuclease VII converts the supercoiled circular DNA directly into linear product, indicating that the two cleavage reactions must occur within the lifetime of the protein-junction complex. By contrast, a heterodimer of active enzyme and an inactive mutant endonuclease VII leads to the formation of nicked circular product, showing that the subunits operate fully independently.
Giraud-Panis
M J
MJ
CRC Nucleic Acid Structure Research Group, Department of Biochemistry, The University, Dundee, UK.
Lilley
D M
DM
D017122:Bacteriophage T4 / Q000201:enzymology; D004278:DNA, Superhelical / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D019281:Dimerization; D004706:Endodeoxyribonucleases / Q000235:genetics / Q000378:metabolism*; D005816:Genetic Complementation Test; D009154:Mutation; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000378:metabolism; D011995:Recombination, Genetic; D013329:Structure-Activity Relationship
D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; C035571:endodeoxyribonuclease VII