Objectif TGF-ß related growth factors signal through Smad proteins that transfer the signal from the receptor directly into the nucleus. The receptor-regulated Smads (R-Smads) Smad2 and SmadS arephosphorylated by the receptor kinase and form complexes with Smad4 that accumulate in the nucleus, where they participate in the regulation of gene transcription. Like Smad4, Smad2 and SmadS exhibit nucleocytoplasmic shuttling irrespective of the presence or absence of a signal.This project aims at answering two clearly define d questions. Firstly, how does phosphorylation of R-Smadstrigger nuclear accumulation despite their constitutive shuttling properties? And secondly, which import and export receptors (karyopherins) or other regulatory proteins are involved in these transport processes? To answer these questions, a set of powerful experimental approaches will be employed. In the first phase, a novel GFP photo-activation approach will allow detailed studies of nucleocytoplasmic transport kinetics in real time in vivo. I will be able to dissect import from export and study both processes independently. In the second phase, I will take advantage of an in vitro import assay using permeabilised cells depleted of cytoplasm to directly compare the import characteristics of recombinant full length phosphorylated, recombinant unphosphorylated Smad2 as well as of active PhosphoSmad2/Smad4 complexes under different conditions. In a more wide-ranging approach, novel players involved in R-Smad import and export will be identified by a screen using a n siRNA library targeting over 8000 human mRNAs.A quantitative in vivo export assay will be used to further characterise the effect of knock down of candidate proteins involved in R-Smad export. The project involves a plethora of novel methods, including cut ting edge microscopy and employment of functional genomics using an RNAi library. Champ scientifique natural sciencesmathematicspure mathematicsmathematical analysisdifferential equationsnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencesdevelopmental biologynatural sciencesphysical sciencesopticsmicroscopyconfocal microscopynatural sciencesmathematicsapplied mathematicsmathematical model Programme(s) FP6-MOBILITY - Human resources and Mobility in the specific programme for research, technological development and demonstration "Structuring the European Research Area" under the Sixth Framework Programme 2002-2006 Thème(s) MOBILITY-2.1 - Marie Curie Intra-European Fellowships (EIF) Appel à propositions FP6-2002-MOBILITY-5 Voir d’autres projets de cet appel Régime de financement IIF - Marie Curie actions-Incoming International Fellowships Coordinateur CANCER RESEARCH UK - LONDON RESEARCH INSTITUTE Contribution de l’UE Aucune donnée Adresse 61 Lincoln's Inn Fields LONDON Royaume-Uni Voir sur la carte Liens Site web Opens in new window Coût total Aucune donnée
