Catalytic DNA Polymerization Can Be Expedited by Active Product Release

The sequence‐specific hybridization of DNA facilitates its use as a building block for designer nanoscale structures and reaction networks that perform computations. However, the strong binding energy of Watson–Crick base pairing that underlies this specificity also causes the DNA dehybridization rate to depend sensitively on sequence length and temperature. This strong dependency imposes stringent constraints on the design of multi‐step DNA reactions. Here we show how an ATP‐dependent helicase, Rep‐X, can drive specific dehybridization reactions at rates independent of sequence length, removing the constraints of equilibrium on DNA hybridization and dehybridization. To illustrate how this new capacity can speed up designed DNA reaction networks, we show that Rep‐X extends the range of conditions where the primer exchange reaction, which catalytically adds a domain provided by a hairpin template to a DNA substrate, proceeds rapidly. The rate of the primer exchange reaction is a strongly peaked function of the binding strength between the reactant and catalyst strands. This trend holds for many catalytic reactions and is known as Sabatier's principle. A helicase that consumes ATP to selectively remove the products from the catalysts can speed up the reaction in the strong‐binding regime and reduce the dependency of the catalytic rate on the binding strength.