Topoisomerase II Is Crucial for Fork Convergence during Vertebrate Replication Termination

Abstract

Summary: Termination of DNA replication occurs when two replication forks converge upon the same stretch of DNA. Resolution of topological stress by topoisomerases is crucial for fork convergence in bacteria and viruses, but it is unclear whether similar mechanisms operate during vertebrate termination. Using Xenopus egg extracts, we show that topoisomerase II (Top2) resolves topological stress to prevent converging forks from stalling during termination. Under these conditions, stalling arises due to an inability to unwind the final stretch of DNA ahead of each fork. By promoting fork convergence, Top2 facilitates all downstream events of termination. Converging forks ultimately overcome stalling independently of Top2, indicating that additional mechanisms support fork convergence. Top2 acts throughout replication to prevent the accumulation of topological stress that would otherwise stall converging forks. Thus, termination poses evolutionarily conserved topological problems that can be mitigated by careful execution of the earlier stages of replication. : To complete DNA synthesis replication, forks must converge on the same stretch of DNA. In vertebrates this process occurs rapidly, but it is unclear which mechanisms support fork convergence. Heintzman et al. find that topoisomerase II promotes fork convergence by preventing accumulation of topological stress earlier during replication. Keywords: DNA replication, termination, topoisomerase, fork stalling, genome stability, Xenopus, chemotherapy, cancer