Reconstitution of a eukaryotic replisome reveals the mechanism of asymmetric distribution of DNA polymerases

Eukaryotes require 3 DNA polymerases for normal replisome operations, DNA polymerases (Pol) alpha, delta and epsilon. Recent biochemical and structural studies support the asymmetric use of these polymerases on the leading and lagging strands. Pol epsilon interacts with the 11-subunit CMG helicase, forming a 15-protein leading strand complex that acts processively in leading strand synthesis in vitro, but Pol epsilon is inactive on the lagging strand. The opposite results are observed for Pol delta with CMG. Pol delta is highly active on the lagging strand in vitro, but has only feeble activity with CMG on the leading strand. Pol a also functions with CMG to prime both strands, and is even capable of extending both strands with CMG present. However, extensive DNA synthesis by Pol a is sharply curtailed by the presence of either Pol epsilon or Pol delta, which limits the role of the low fidelity Pol a to the initial priming of synthesis.