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Content archived on 2024-05-29

Role of the plant DDB1 complex in chromatin remodelling at loci of UV damaged DNA and developmental genes

Objective

Research on DNA repair in animals and on photomorphogenesis in plants led to the discovery of common factors, the DNA Damage Binding complex formed by DDB1 and associated proteins, which associates with chromatin remodelling and ubiquitination activities.

In mammals this complex is tightly bound to chromatin upon UV irradiation and could participate in early steps to make the DNA lesions more accessible to the repair machinery. A plant-specific CDD complex containing proteins COP10, DDB1 and DET1 has been recently discovered, exhibits similar ubiquitination activities and binds nonacetylated histone H2B.

We will investigate the multifaceted roles that this complex may play in plant cells to interface protein ubiquitination, chromatin remodelling, DNA repair and the regulation of light-responsive genes at a global level during de-etiolation. Purification of the complex from dark- or light-grown seedlings, and irradiation of mononucleosomes with UV prior to binding assays will allow to determine the relative af finity of the plant CDD complex for these substrates.

Analysis of the bound histones and of their ubiquitination and acetylation states will be performed. CDD function will also be assessed using a plant in vitro DNA repair system recently established in t he host laboratory, focusing on the possible requirement of the CDD activity in mono- or polyubiquitination to efficiently repair naked or chromatin-packaged DNA in vitro.

Its genomic targets will be identified by ChIP, compared to the repertoire of genes known to be misregulated in photomorphogenic mutants and their methyl-cytosine content will be examined. In complement, FRET and other in vivo approaches will be used to investigate the CDD complex association with euchromatin and transcriptionaly-inactive heterochromatin.

This project contributes to the current strong interest in studying epigenetic controls in plants and would allow investigating a challenging new field of gene regulatory mechanisms.

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Keywords

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Topic(s)

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Call for proposal

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FP6-2004-MOBILITY-5
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Funding Scheme

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EIF - Marie Curie actions-Intra-European Fellowships

Coordinator

STAZIONE ZOOLOGICA ANTON DOHRN
EU contribution
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Total cost

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