Geometric arrangements of Tn3 resolvase sites.
Site-specific recombination by Tn3 resolvase normally occurs in vitro and in vivo only between directly repeated res sites on the same supercoiled DNA molecule. However, with multiply interlinked catenane substrates consisting of two DNA rings each containing a single res site, resolvase efficiently carried out intermolecular recombination. The topology of the knots produced by several rounds of this reaction proves that the DNA within the synaptic intermediate is coiled in an interwound (plectonemic) fashion rather than wrapped solenoidally around resolvase as in previously characterized supercoiled DNA-protein complexes. The synaptic intermediate can contain equivalently supercoil, catenane, or knot crossings as long as the res sites have a right-handed coiling and a particular relative orientation. The structure of the product knots and catenanes also shows the path the DNA takes during strand exchange. Intermolecular recombination within multiply linked catenanes required negative supercoiling, as does the standard intramolecular reaction.
Benjamin
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Department of Molecular and Cell Biology, University of California, Berkeley 94720.
Cozzarelli
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D004264:DNA Topoisomerases, Type I / Q000378:metabolism; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics; D008961:Models, Structural; D009690:Nucleic Acid Conformation; D009713:Nucleotidyltransferases / Q000378:metabolism*; D010957:Plasmids; D015183:Restriction Mapping; D019895:Transposases
D004251:DNA Transposable Elements; D009713:Nucleotidyltransferases; D019895:Transposases; D004264:DNA Topoisomerases, Type I