Description du projet
Méthylation de l’ADN: séquence d’ADN, densité de méthylation et liaison du facteur de transcription
Les changements au niveau de l’expression de certains gènes à court et à long terme jouent un rôle crucial dans la santé et la maladie. La méthylation, à savoir l’ajout d’un groupement méthyle à l’ADN, est un mécanisme clé de régulation de l’expression génétique. Il empêche l’expression des gènes en inhibant la liaison des facteurs de transcription; ses mécanismes demeurent toutefois mal compris. Le projet ReaDMe, financé par le Conseil européen de la recherche, identifiera les régulateurs transcriptionnels qui répondent à la méthylation de l’ADN in vivo grâce à une combinaison d’outils de génomique, d’édition du génome et de protéomique dans les cellules embryonnaires et somatiques. Les connaissances acquises contribueront au développement d’un dispositif intégré permettant d’identifier et de caractériser le site du génome où la méthylation de l'ADN influence les profils de liaison des facteurs de transcription in vivo.
Objectif
DNA and chromatin modifications are essential for proper control of gene expression during development. How these marks alter transcriptional programs and modulate binding patterns of sequence specific transcription factors (TF) remains poorly understood. This currently limits our interpretation of epigenomic maps towards their incorporation into predictive models of gene regulation.
ReaDMe has the ambitious goal to systematically define the sensitivity of TFs to local levels of DNA methylation in vivo. We will use a combination of genomics, genome editing and proteomics tools to comprehensively identify transcriptional regulators that respond to DNA methylation. As a first approach, we will interrogate changes in the global TF binding landscape when DNA methylation is ablated from the genome. Using both embryonic stem cells and somatic cells, these experiments are aimed at identifying sites that are occupied by TFs in a DNA methylation dependent manner within different cellular context. Secondly, we will combine parallelized chromosomal insertions with targeted footprinting to determine the link between DNA sequence context, methylation density and TF binding. In a third approach we will define the global chromatin proteome as a function of DNA methylation. Through the use of a novel and orthogonal proteomics assay, we will characterize DNA methylation sensitive changes in the chromatin-bound proteome. Candidate factors predicted from all approaches will be validated and functionally characterized through direct genome-wide mapping as well as loss of function analysis.
ERC funding would enable ReaDMe to develop an integrated setup to in vivo identify and characterize where DNA methylation influences the cis-regulatory landscape by modulating binding profiles of trans-acting factors. This goal represents a crucial step towards comprehensive understanding of the genomic readout of DNA methylation and its impact on gene regulation.
Champ scientifique
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteinsproteomics
- natural sciencesbiological sciencesgeneticsDNA
- medical and health sciencesmedical biotechnologycells technologiesstem cells
- natural sciencesbiological sciencesgeneticsgenomes
- natural sciencesbiological sciencesgeneticsepigenetics
Programme(s)
Thème(s)
Régime de financement
ERC-ADG - Advanced GrantInstitution d’accueil
4058 Basel
Suisse