Obiettivo Mapping and sequencing of individual human genes have advanced at a rapid pace. In order to fully understand gene regulation and safely control the expression of important human disease related genes in somatic gene therapy, it is not sufficient to know the regulatory and coding sequences of each gene and of individual factors involved in their expression. It is equally essential to understand the dynamic, three-dimensional nuclear organization and compartmentalization of higher order chromatin structures in relation to other nuclear domains representing the machineries for transcription, splicing, replication and repair. No single laboratory has the intellectual and methodological scope to tackle this problem on its own. In this proposal the expertise required for an interdisciplinary and integrated European effort is brought together by 10 laboratories from 5 European countries. We will combine multi-colour fluorescence in situ hybridization (FISH) of chromatin structures (from the level of individual chromatin loop domains to the level of entire chromosome territories) together with the immunocytochemical localization of essential protein complexes, as well as in situ assays of nascent RNA synthesis. 3D-analyses will be undertaken at the light microscopic level, including newly developed microscope types with improved resolution and advanced 3D-image analysis procedures. 3D electron microscopic studies will supplement these light microscopic investigations. At the level of entire chromosomes we will perform measurements of the volume, shape and surface of their respective interphase territories and study the 3D-positioning of centromeric and telomeric subregions, as well as of selected individual genes. Model studies of normal human cell nuclei include the overall organization of the active and inactive X-chromosome territories and the three-dimensional topology of the major histocompatibility complex (MHC) in relation to the 3D-distribution of nascent RNA, replication clusters and specific protein domains. For comparison, the 3D-topology of aberrant higher order chromatin structures, such as homogeneously stained regions (HSR) and double minute chromosomes (DMC), will be studied in tumour cell nuclei. The applicants expect that their combined efforts will (i) establish tools for 3D-gene mapping and 3D-genome analysis and (ii) provide a better understanding of the influence of higher order chromatin structures in situ on gene regulation and function in human normal and tumour cell nuclei. Such an approach will contribute to provide the necessary basis for an understanding of the topology and dynamics of specific, nuclear 3D-structures during development, differentiation and malignancy. Campo scientifico natural sciencesmathematicspure mathematicstopologynatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesphysical sciencesopticsmicroscopynatural sciencesbiological sciencesgeneticsRNAnatural sciencesbiological sciencesgeneticschromosomes Programma(i) FP4-BIOMED 2 - Specific research, technological development and demonstration programme in the field of biomedicine and health, 1994-1998 Argomento(i) 5.2 - Analysis of gene function and interaction Invito a presentare proposte Data not available Meccanismo di finanziamento CSC - Cost-sharing contracts Coordinatore Ruprecht-Karls-Universität Heidelberg Contributo UE Nessun dato Indirizzo 10/I,Richard-Wagner-Strasse 80333 München Germania Mostra sulla mappa Costo totale Nessun dato Partecipanti (6) Classifica in ordine alfabetico Classifica per Contributo UE Espandi tutto Riduci tutto Centre National de la Recherche Scientifique Francia Contributo UE Nessun dato Indirizzo 2,Place Jussieu 75251 Paris Mostra sulla mappa Costo totale Nessun dato European Molecular Biology Laboratory Germania Contributo UE Nessun dato Indirizzo Meyerhofstraße 1 69012 Heidelberg Mostra sulla mappa Costo totale Nessun dato Imperial Cancer Research Fund (ICRF) Regno Unito Contributo UE Nessun dato Indirizzo 44 Lincoln's Inn Fields WC2A 3PX London Mostra sulla mappa Costo totale Nessun dato Universiteit van Amsterdam Paesi Bassi Contributo UE Nessun dato Indirizzo 12,Plantage Muidergracht 1018 TV Amsterdam Mostra sulla mappa Costo totale Nessun dato Université Joseph Fourier Francia Contributo UE Nessun dato Indirizzo Domaine Universitaire 38041 Grenoble Mostra sulla mappa Costo totale Nessun dato Université de Lausanne Svizzera Contributo UE Nessun dato Indirizzo 27,Rue du Bugnon 1005 Lausanne Mostra sulla mappa Costo totale Nessun dato
