H2020Индивидуална стипендия2017–2019

Rep. Fork Restart · Restart of blocked replication fork at replication fork barrier

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

Период: 2017-05-01 → 2019-04-30
Финансиране от ЕС: 183 455 €
Участници: 1
Схема: MSCA-IF-EF-ST

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Накратко на български

Механизмите за рестартиране на блокираното копиране на ДНК се анализират чрез използването на специален протеинов бариер. Разбирането на този процес помага да се обясни генетичната нестабилност, която води до рак и неврологични заболявания.

Този кратък обзор е генериран от изкуствен интелект

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

Резултати накратко

Restart of blocked replication fork at replication fork barrier

Each human cell is created after genetic information stored in DNA, which contains 12 billion DNA bases. These have to be maintained intact the whole life and they have to be properly copied during development. Even an adult has to replicate billions of bases every day (e.g. for cell renewal in intestine or skin). However, replication is constantly perturbed, and errors occur. This can drive evolution, but at the same time replication errors cause genetic instability, which can lead to numerous diseases like cancer, and numerous neurological, neurodegenerative and neuromuscular disorders. My research proposal addressed a specific mechanism of replication-dependent genetic instability. Replication forks (RFs) copy all DNA and they can get stalled by different obstacles like for example DNA-bound proteins (as in Fig.1). The RF encountering a barrier on DNA might be resolved by one of the following three strategies: (i) Stabilisation of stalled RFs allows to wait for the rescue by (ii) an adjacent converging RF. Low numbers of converging RF are present in common fragile sites, which are frequent sites of rearrangement in cancer, and are particularly sensitive. (iii) Therefore, cells have developed a mechanism to form a new RF and restart the stalled RF (Fig.1: Scheme showing how stalled replication fork can be resolved). The restart represents a solution for these regions, but at a cost. The restarted RF is error prone, but the cell does not lose precious genetic information as we showed in Marie Curie funded project. We used a model protein barrier (called “RTS1”), which leads to the restart of the stalled RF. The overall objective was to understand how the cell deals with such a challenging situation. How does it restart? What precedes the restart? Where is the restarted RF formed? How far can the restarted RF travel? The data in the project contributes to our knowledge how cells cope with fragile and difficult to replicate regions. This will ultimately allow the design of a new generation of drugs which can be exploited in cancer treatment as well as in prevention to several diseases.

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

Цел на проекта

Errors in replication of DNA lead to serious diseases like cancer, inherited neurological and muscular diseases. Replication is constantly challenged by various obstacles that can result in error-prone replication. Replication fork barriers (RFBs) include proteins bound to DNA, structure forming sequences and DNA/RNA hybrids. Replication forks (RFs) stalled or blocked at these barriers can be rescued by a converging fork. But if this fails blocked RF must be restarted to complete replication. Restart using homologous recombination (HR) can lead to gross chromosomal rearrangements and copy number variations. The newly restarted replication fork is also prone to slippage leading to mutation and rearrangements. The research questions I will address are how this RF is restarted and why it is error prone. I will focus on restart at a site-specific RFB in fission yeast, RTS1. Restart at RTS1 has been well characterised in the host laboratory. My project will identify DNA structures formed during the RF restart and characterise recruitment of factors involved in the restart of the collapsed RF. The acquired data will enable us to further understand the mechanism of RF restart and to unravel the cause of the error prone nature of newly restarted RF. I consider this fellowship a key step in my career as it allows me to transit from the analysis of site-specific lesions in prokaryotes (previous doctoral fellowship) to eukaryotic cells, gain experience in a wide range of techniques (2D gels, Electron microscopy, ChIP) and build contacts with experts in the field of eukaryotic replication restart. This is necessary for my future independent research and will enable me to set up my lab to study molecular actors and DNA structures during bypass of single lesions in bacteria and eukaryotic cells. The mentoring during the fellowship will provide me with the necessary training to successfully establish myself as an independent investigator.

Оригинален текст от CORDIS (на английски).

Участници

THE UNIVERSITY OF SUSSEX · BrightonКоординаторОбединеното кралство