A model for topoisomerase I-mediated insertions and deletions with duplex DNA substrates containing branches, nicks, and gaps.

摘要

The ability of DNA topoisomerase I to promote insertions and deletions in vitro has been studied at nucleotide resolution for structurally diverse DNA substrates that uncouple the cleavage and ligation reactions of the enzyme. Topoisomerase I-mediated ligations afforded DNA duplexes having deletions and insertions with "branched" substrates and deletions up to 18 nucleotides in length with substrates containing nicks or gaps. In addition, a number of the acceptor substrates altered the preferred site of DNA cleavage, thereby increasing the diversity of accessible ligation products. Also demonstrated by the production of two "recombinant" duplexes from a single set of reactants was the potential for amplification of such alterations. These findings illustrate plausible mechanisms by which topoisomerase I-mediated illegitimate recombination may obtain at a molecular level.