MONITORING THE CHANGES OF MYOD INTERACTION WITH TRANSCRIPTIONAL CO-ACTIVATORS AND CO-REPRESSORS AND THEIR SUB-NUCLEAR LOCALIZATION DURING THE MYOGENIC PROGRAM

This research is dedicated to the identification of molecular targets for pharmacological manipulation of the myogenic program, a critical issue in both setting up therapeutical interventions for neuromuscolar diseases and enhancing the efficiency of muscle-mediated gene therapy. In this project are proposed experiments aimed at addressing the spatial and temporal dynamic of changes of protein complexes containing MyoD with either co-repressors or co-activators. These experiments will be performed in both established myogenic cell lines and primary coltures of satellite muscle cells, by using FRET analysis to monitor protein-protein interactions in living cells and co-localization studies by immunofluoresce deconvolution analysis to investigate changes in sub-nuclear redistribution of endogenous complexes implicated in the control of muscle specific transcription. This analysis will be conducted under different conditions, such as proliferation of undifferentiated myoblasts, their spontaneous differentiation, or in the presence of agents which promote or inhibit differentiation. The results gathered from this study may provide important information relative to the dynamic of formation and spatial distribution of nuclear protein complexes that regulate muscle-specific transcription and hence the myogenic program.