Objective 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. Fields of science 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) Topic(s) ERC-AG-ID1 - ERC Advanced Grant Interdisciplinary Panel Call for proposal ERC-2010-AdG_20100224 See other projects for this call Funding Scheme ERC-AG - ERC Advanced Grant Host institution CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS EU contribution € 1 559 647,20 Address RUE MICHEL ANGE 3 75794 Paris France See on map Region Ile-de-France Ile-de-France Paris Activity type Research Organisations Administrative Contact Julie Zittel (Ms.) Principal investigator Vincent,jean,marie,christian Croquette (Dr.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data Beneficiaries (3) Sort alphabetically Sort by EU Contribution Expand all Collapse all CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS France EU contribution € 1 559 647,20 Address RUE MICHEL ANGE 3 75794 Paris See on map Region Ile-de-France Ile-de-France Paris Activity type Research Organisations Administrative Contact Julie Zittel (Ms.) Principal investigator Vincent,jean,marie,christian Croquette (Dr.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data UNIVERSITAT DE BARCELONA Spain EU contribution € 466 240,80 Address GRAN VIA DE LES CORTS CATALANES 585 08007 Barcelona See on map Region Este Cataluña Barcelona Activity type Higher or Secondary Education Establishments Administrative Contact Xavier Gutierrez (Mr.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data THE PENNSYLVANIA STATE UNIVERSITY United States EU contribution € 167 678,00 Address TECHNOLOGY CENTER BUILDING 110 16802 7000 UNIVERSITY PARK PA See on map Activity type Higher or Secondary Education Establishments Administrative Contact Lynn P. Mitchell (Mr.) Links Contact the organisation Opens in new window Website Opens in new window Total cost No data