Resolvase-catalysed reactions between res sites differing in the central dinucleotide of subsite I.
The resolvase-catalysed reaction between two res sites in a circular DNA substrate normally gives two circular recombination products linked in a two-noded catenane. Homology between the two res sites at the central overlap dinucleotide of subsite I is important for recombination. Reactions between res sites differing at one position in the central dinucleotide (AC X AT) gave a low yield of recombinants containing mismatched base-pairs, but gave large amounts of a non-recombinant four-noded knot. This result was predicted by a 'simple rotation' model for strand exchange. The mismatch is evidently recognized only after commitment to an initial 180 degrees rotation of the resolvase-linked DNA ends, and it induces a second 180 degrees rotation which restores correct base-pairing at the overlap, giving the four-noded product. Correct base-pairing is not essential for religation, but may be important for release of the products. Characteristic patterns of 4, 6, 8 and 10 node knots, or 4, 8, 12 and 16 node knots were obtained, depending on the reaction conditions and the resolvase. Two pathways for multiple rounds of rotation in 360 degrees steps are inferred. The results support a model for strand exchange by supercoil-directed subunit rotation within a resolvase tetramer.
Stark
W M
WM
Institute of Genetics, University of Glasgow, UK.
Grindley
N D
ND
Hatfull
G F
GF
Boocock
M R
MR
D001483:Base Sequence; D002384:Catalysis; D004247:DNA / Q000235:genetics / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D006026:Glycoside Hydrolases / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009713:Nucleotidyltransferases / Q000378:metabolism*; D010957:Plasmids; D011995:Recombination, Genetic; D013379:Substrate Specificity; D019895:Transposases; D043324:beta-Fructofuranosidase
D004247:DNA; D009713:Nucleotidyltransferases; D019895:Transposases; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase