Obiettivo 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. Campo scientifico natural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsenzymes Programma(i) 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) Argomento(i) ERC-AG-ID1 - ERC Advanced Grant Interdisciplinary Panel Invito a presentare proposte ERC-2010-AdG_20100224 Vedi altri progetti per questo bando Meccanismo di finanziamento ERC-AG - ERC Advanced Grant Istituzione ospitante CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS Contributo UE € 1 559 647,20 Indirizzo RUE MICHEL ANGE 3 75794 Paris Francia Mostra sulla mappa Regione Ile-de-France Ile-de-France Paris Tipo di attività Research Organisations Contatto amministrativo Julie Zittel (Ms.) Ricercatore principale Vincent,jean,marie,christian Croquette (Dr.) Collegamenti Contatta l’organizzazione Opens in new window Sito web Opens in new window Costo totale Nessun dato Beneficiari (3) Classifica in ordine alfabetico Classifica per Contributo UE Espandi tutto Riduci tutto CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS Francia Contributo UE € 1 559 647,20 Indirizzo RUE MICHEL ANGE 3 75794 Paris Mostra sulla mappa Regione Ile-de-France Ile-de-France Paris Tipo di attività Research Organisations Contatto amministrativo Julie Zittel (Ms.) Ricercatore principale Vincent,jean,marie,christian Croquette (Dr.) Collegamenti Contatta l’organizzazione Opens in new window Sito web Opens in new window Costo totale Nessun dato UNIVERSITAT DE BARCELONA Spagna Contributo UE € 466 240,80 Indirizzo GRAN VIA DE LES CORTS CATALANES 585 08007 Barcelona Mostra sulla mappa Regione Este Cataluña Barcelona Tipo di attività Higher or Secondary Education Establishments Contatto amministrativo Xavier Gutierrez (Mr.) Collegamenti Contatta l’organizzazione Opens in new window Sito web Opens in new window Costo totale Nessun dato THE PENNSYLVANIA STATE UNIVERSITY Stati Uniti Contributo UE € 167 678,00 Indirizzo TECHNOLOGY CENTER BUILDING 110 16802 7000 UNIVERSITY PARK PA Mostra sulla mappa Tipo di attività Higher or Secondary Education Establishments Contatto amministrativo Lynn P. Mitchell (Mr.) Collegamenti Contatta l’organizzazione Opens in new window Sito web Opens in new window Costo totale Nessun dato