Mechanism and structure of the human DNA replication machinery
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Accurate and timely chromosome replication is essential to preserve genome integrity. The cellular machinery responsible for this process is called the replisome. The eukaryotic replisome is a large, sophisticated and highly regulated molecular machine that possesses all of the activities required for rapid and efficient chromosome replication. Replisomes must synthesise vast quantities of DNA, whilst keeping mistakes to an absolute minimum. To illustrate the scale of the challenge, you will likely synthesise in excess of 100,000 kilometres of DNA whilst reading this summary.
As a post doc, I reconstituted for the first time the regulated assembly of a eukaryotic replisome with purified proteins from budding yeast1. The development of this system marked the beginning of a new era in the study of eukaryotic chromosome replication. This has enabled us to address important questions in ways that were not previously possible, including: how the yeast replisome achieves rapid and efficient DNA replication2, how it navigates chromatin templates3, and how it responds to DNA template damage4. In contrast to the progress made with the yeast replisome, at present, a comparable system using purified human proteins is yet to be established.
We are looking for an ambitious and enthusiastic student to join our research group. During your PhD we will seek to develop a reconstituted system to assemble a functional replisome with purified human proteins, which we will use for mechanistic and structural analysis. You will become an expert in protein purification and complex biochemical reconstitution, whilst also having the opportunity to gain experience in structural biology. Our group provides an excellent environment to learn new techniques, both directly from myself and from experienced post docs in the lab, whilst working on an independent project.
- Yeeles, J.T., Deegan, T.D., Janska, A., Early, A. and Diffley, J.F. (2015)
Regulated eukaryotic DNA replication origin firing with purified proteins.
Nature 519(7544): 431-435
- Yeeles, J.T.P., Janska, A., Early, A., and Diffley, J.F.X. (2017).
How the Eukaryotic Replisome Achieves Rapid and Efficient DNA Replication.
Mol Cell 65, 105-116.
- Kurat, C.F., Yeeles, J.T.P., Patel, H., Early, A., and Diffley, J.F.X. (2017).
Chromatin Controls DNA Replication Origin Selection, Lagging-Strand Synthesis, and Replication Fork Rates.
Mol Cell 65, 117-130.
- Taylor, M.R.G., and Yeeles, J.T.P. (2018).
The Initial Response of a Eukaryotic Replisome to DNA Damage.
Mol Cell 70, 1067-1080 e1012.