Eukaryotic replication

Our understanding of DNA replication has made gigantic leaps forward since the discovery of the double helical form of DNA by Watson and Crick in 1952. We know many of the structures and functions of the proteins and enzymes involved. Many of these discoveries have been made by studying DNA replication in simple systems, such as viruses or bacteria. These continue to yield valuable insights, but recently, advances in the reconstitution of the yeast replisome have made it possible to gain insights into eukaryotic replication.

Here we take a multi-pronged approach based on single-molecule investigations to studying eukaryotic replication. We employ magnetic tweezers to study a relatively simple replicative helicase from simian virus that is capable of unwinding DNA ahead of a eukaryotic replisome. And we study the dynamics of replication in the yeast replisome using a combination of magnetic tweezers and single-molecule fluorescence approaches. Currently, in the latter system we focus on examining the influence of physical parameters such as torque and twist on several stages of replication.

Researchers currently involved in the project

  • Mariana Koeber
  • Richard Janissen
  • Humberto Sanchez
  • Kaley McCluskey
  • Theo van Laar

Collaborations

  • The Xiaojiang Chen Lab (University of Southern California, USA)
  • The John Diffley Lab (Francis Crick Institute, UK)

Publications specific to this project

B.A. Berghuis, M. Koeber, T. van Laar, and N.H. Dekker
High-throughput, high-force probing of DNA-protein interactions with magnetic tweezers
Methods, online publication March 30 (2016) PDF