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The role of the iron-sulpur cluster in human DNA polymerase delta

Objective

The human genome is composed of 3 billion base pairs of DNA that are replicated prior to every cell division with an astonishing accuracy. High-fidelity replication is necessary to maintain genome stability, and hence to avoid premature ageing and cancer. This project aims at understanding the role of the iron-sulphur (FeS) cluster in human DNA polymerase delta (Pol δ), one of the major replicases.
FeS clusters are ancient and versatile co-factors that are commonly known for their function in electron transport in the mitochondrial respiratory chain. In recent years, a surprising number of proteins involved in DNA metabolism have been discovered to contain an FeS cluster including all replicative DNA polymerases in yeast. While the requirement of an FeS cluster for the function of replicases was demonstrated, the actual role of the FeS cluster in these enzymes has remained largely elusive.
The TRISCPOL project aims to: 1) Confirm and characterise the FeS cluster in human Pol δ by using iron incorporation assays, UV-visible and electron paramagnetic resonance spectroscopy. 2) Determine the role of the FeS cluster in human Pol δ in vitro. Purified Pol δ will be subjected to oxidative stress conditions, and then changes in structural and functional features will be measured. A combination of techniques including deuterium-hydrogen exchange mass spectrometry, as well as DNA binding, DNA synthesis, processivity and fidelity assays, will be employed. 3) Define the role of the FeS cluster in human Pol δ in vivo. Mutants in the FeS cluster-binding region will be engineered and tested for their ability to functionally complement cells depleted of endogenous Pol δ.
Unveiling the role of the FeS cluster in human Pol δ will shed new light on the principle of eukaryotic DNA replication. Moreover, this knowledge will contribute to our understanding of the molecular basis of cancer and may eventually allow the development of novel strategies of treatment.

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MSCA-IF-EF-ST - Standard EF

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2015

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Coordinator

UNIVERSITAT ZURICH
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 187 419,60
Address
RAMISTRASSE 71
8006 Zurich
Switzerland

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Region
Schweiz/Suisse/Svizzera Zürich Zürich
Activity type
Higher or Secondary Education Establishments
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Total cost

The total costs incurred by this organisation to participate in the project, including direct and indirect costs. This amount is a subset of the overall project budget.

€ 187 419,60
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