H2020Индивидуална стипендия2016–2018

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

„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“

Период: 2016-05-01 → 2018-04-30
Финансиране от ЕС: 187 420 €
Участници: 1
Схема: MSCA-IF-EF-ST

Линиите свързват координатора с партньорите.

Накратко на български

Железните серни клъстери в човешкия ензим ДНК полимераза делта се изследват, за да се разбере как те влияят върху сглобяването на протеина. Това помага за по-доброто разбиране на механизмите, които водят до нестабилност на генома и развитие на рак.

Този кратък обзор е генериран от изкуствен интелект

Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.

Резултати накратко

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

According to official reports released by the European Parliament Commission, it is estimated that 1.2 million of Europeans died of cancer in 2012. The faithful duplication of the genome prior to each cell division (DNA replication) is one of the most essential cellular processes. Deregulation of DNA replication is known to be one of the cellular mechanisms that contribute to genome instability and ultimately can cause cancer. Therefore, revealing the mechanisms of human DNA replication is an important step towards a comprehensive understanding of the cellular processes underlying cancer aetiology and malignant transformation. Eukaryotic DNA replication is performed through the collaborative effort of the three replicative DNA polymerases Pol α, Pol δ and Pol ε. A recent study in yeast suggests a pivotal role of Pol δ in DNA replication. Additionally, Pol δ activity is also implicated in DNA repair, DNA damage tolerance and homologous recombination. Iron-sulphur (FeS) clusters are ancient partners in the origin of life that predate cells, acetyl-CoA metabolism, DNA and the RNA world. In recent years, a surprising number of proteins involved in DNA metabolism have been discovered to contain this inorganic cofactor. Intriguingly, all replicative DNA polymerases and DNA primase in yeast coordinate an FeS cluster. Most of our understanding of human DNA replication as a process has been extrapolated from research on model organisms. The studies on the role of the FeS cluster in yeast Pol δ clearly demonstrated the requirement of this cofactor for the stable assembly of the holo-enzyme. The objective of this project was to determine the role of the FeS cluster in human Pol δ. First, I confirmed experimentally that human Pol δ – like its yeast homologue – binds to an FeS cluster. Second, I characterised the role of the FeS cluster in human Pol δ in vitro by employing a variety of biochemical techniques. Finally, I studied the role of the FeS cluster in human Pol δ in vivo by using a combination of protein engineering, molecular and cell biology methods.

Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз

Цел на проекта

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.

Оригинален текст от CORDIS (на английски).

Участници

UNIVERSITAT ZURICH · ZurichКоординаторШвейцария