Objectif We propose to investigate the enzymes responsible for DNA replication and repair in micromanipulation experiments with a resolution of a single base. The detailed mechanism by which DNA is synthesized base after base and the coordination of the enzymes involved in this process are not fully understood. We shall develop new magnetic tweezers using lithographic techniques associated with evanescent field detection to address these issues. We shall build arrays of these devices working in parallel, each one on a single DNA molecule and where the measurement of its extension reveals enzymatic activity. The DNA molecule in these devices will form a hairpin the opening of which can be detected with a single base resolution.We will study the different enzymes involved in DNA replication. Firstly, we wish to follow in real time the incorporation of bases one by one by a DNA-polymerase and investigate the proof-reading mechanism of this enzyme. We shall also investigate the translocation mechanisms of different helicases involved in DNA replication and repair. Finally, we plan to study the cooperative action between different enzymes involved in the replication machinery with the help of parallelized micro-tweezers: the coupling between helicase and primase in the lagging strand synthesis, the coupling between the helicase and polymerase during leading strand synthesis and the coordination between leading and lagging strand synthesis.Moreover observing a DNA-polymerase at the single base level is the first step of a DNA sequencing method. Preliminary experiments demonstrate that the unzipping assay is a new way to determine the position of a small DNA sequence with single base resolution. We shall investigate different experimental schemes to achieve this goal. Champ scientifique natural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes Programme(s) FP7-IDEAS-ERC - Specific programme: "Ideas" implementing the Seventh Framework Programme of the European Community for research, technological development and demonstration activities (2007 to 2013) Thème(s) ERC-AG-ID1 - ERC Advanced Grant Interdisciplinary Panel Appel à propositions ERC-2010-AdG_20100224 Voir d’autres projets de cet appel Régime de financement ERC-AG - ERC Advanced Grant Institution d’accueil CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS Contribution de l’UE € 1 559 647,20 Adresse RUE MICHEL ANGE 3 75794 Paris France Voir sur la carte Région Ile-de-France Ile-de-France Paris Type d’activité Research Organisations Contact administratif Julie Zittel (Ms.) Chercheur principal Vincent,jean,marie,christian Croquette (Dr.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée Bénéficiaires (3) Trier par ordre alphabétique Trier par contribution de l’UE Tout développer Tout réduire CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS France Contribution de l’UE € 1 559 647,20 Adresse RUE MICHEL ANGE 3 75794 Paris Voir sur la carte Région Ile-de-France Ile-de-France Paris Type d’activité Research Organisations Contact administratif Julie Zittel (Ms.) Chercheur principal Vincent,jean,marie,christian Croquette (Dr.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée UNIVERSITAT DE BARCELONA Espagne Contribution de l’UE € 466 240,80 Adresse GRAN VIA DE LES CORTS CATALANES 585 08007 Barcelona Voir sur la carte Région Este Cataluña Barcelona Type d’activité Higher or Secondary Education Establishments Contact administratif Xavier Gutierrez (Mr.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée THE PENNSYLVANIA STATE UNIVERSITY États-Unis Contribution de l’UE € 167 678,00 Adresse TECHNOLOGY CENTER BUILDING 110 16802 7000 UNIVERSITY PARK PA Voir sur la carte Type d’activité Higher or Secondary Education Establishments Contact administratif Lynn P. Mitchell (Mr.) Liens Contacter l’organisation Opens in new window Site web Opens in new window Coût total Aucune donnée
