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Human DNA replication with purified proteins

Laboratory of Molecular Biology

Thursday, December 03, 2020 Competition Funded PhD Project (Students Worldwide)

About the Project

Accurate genome duplication is essential for all forms of life. The molecular machine that orchestrates this process is the replisome: a complex ensemble of proteins that co-ordinates DNA template unwinding, DNA synthesis, histone transfer and several other replication-coupled processes. Our laboratory uses a combination of biochemical reconstitution and structural biology to examine replisome structure and function. While I was a postdoc, I reconstituted regulated eukaryotic DNA replication for the first time using purified proteins from budding yeast. This system has enabled us to address several important questions including how leading-strand replication is started and how the replisome responds to DNA template damage. We are currently working towards reconstituting a human replisome entirely with purified proteins. This will enable us to investigate how the human replisome interfaces with DNA repair and recombination pathways that are required to sustain DNA replication under conditions of replication stress that can impede replisome progression. This is important because there are many factors required to maintain replication fork progression in human cells that are absent from budding yeast.

We are looking for an enthusiastic and ambitious student to join our research group. During your PhD you will further develop a reconstituted replication system to investigate how the human replisome collaborates with DNA repair and recombination pathways to sustain replisome progression under stress conditions. You will become an expert in protein purification and complex biochemical reconstitution and will also have the opportunity to gain experience in structural biology. Our focused research group provides an excellent environment to learn new techniques whilst working on an independent research project.

Funding Notes


Taylor, M. R. G. & Yeeles, J. T. P. (2018). The initial response of a eukaryotic replisome to DNA damage. Mol Cell 70, 1067-1080 e1012.
Bartetić, D., Jenkyn-Bedford, M., Aria, A., Cannone, G., Skehel, M., Yeeles, J.T.P. (2020) Cryo-EM structure of the fork protection complex bound to CMG at a replication fork. Mol Cell 78, 926-940 e13.
Guilliam, T and Yeeles J.T.P. (2020) Reconstitution of translesion synthesis reveals a mechanism of eukaryotic DNA replication restart. Nat Struct Mol Biol 27, 450-460.

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