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Mechanism of nucleosome assembly during DNA replication

Project description

Understanding how post-translational modifications of histones get passed on

Nucleosomes are DNA coiled around histones (proteins). They are the way in which DNA is packaged to fit into the cell's nucleus and, in repeating units, look like beads on a string under a microscope. This complex of DNA and proteins (chromatin) relaxes during transcription to provide transcription factors and other DNA binding proteins access to the DNA. Epigenetic modifications to histone proteins such as methylation can affect gene expression. This, in turn, plays a role in cell fate decisions, tissue development, and immunity. Proper replication of these post-translational modifications along with the DNA prior to cell division is critical, yet mechanisms are largely unknown. EpiRep is studying nucleosome assembly during DNA replication to shed light on this important process.

Objective

Proper inheritance of the epigenetic information during cell division controls cell fate decisions, tissues homeostasis and development, ensuring disease avoidance. We have little understanding however of the mechanisms by which epigenetic information, specifically histone post-translational modifications in nucleosomes, are replicated in parallel to the DNA prior to cell division. This process strictly depends on proper nucleosome formation on the newly synthesized DNA strands. Here, I will study the molecular mechanism of nucleosome assembly during DNA replication. Nucleosome dynamics during DNA replication is controlled by an interconnected network of histone chaperones that converges on the key Chromatin Assembly Factor 1 (CAF-1). I recently elucidated the molecular mechanism of CAF-1-mediated nucleosome assembly, in absence of any other replication components. I developed a quantitative (NAQ) assay that allows, for the first time, the quantification of nucleosome assembly activity in vitro. In cells, CAF-1 is recruited to replication forks by the DNA polymerase processivity factor PCNA. Here, I will capitalize and expand on the assays above to integrate structural data, quantitative biochemical and biophysical measurements, and functional analyses, to elucidate how CAF-1 crosstalks to PCNA, DNA polymerases and other components of the DNA replication machinery in S phase. Specifically, the proposed research will 1) uncover how CAF-1 recruitment by PCNA affects its function in nucleosome assembly, and 2) examine how CAF-1 activity is regulated during ongoing DNA replication. This work will reveal the mechanism of nucleosome assembly during DNA replication and its interplay with S phase signaling. A mechanistic understanding of this pathway will uncover the fundamental principles that control genome and epigenome stability, thus cell fate decisions and disease avoidance.

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Topic(s)

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ERC-STG - Starting Grant

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Call for proposal

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(opens in new window) ERC-2019-STG

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Host institution

KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN - KNAW
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.

€ 1 500 000,00
Address
KLOVENIERSBURGWAL 29 HET TRIPPENHUIS
1011 JV AMSTERDAM
Netherlands

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Total cost

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€ 1 500 000,00

Beneficiaries (1)

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