Descrizione del progetto
La scoperta dell’ordine nascosto nel disordine apparente può stimolare lo sviluppo mirato di farmaci
Molti disturbi respiratori e una serie di malattie infantili comuni quali il morbillo e la parotite sono dovuti a paramixovirus. Negli Stati Uniti, il virus respiratorio sinciziale umano (RSV) è la principale causa di ospedalizzazione infantile. Lo sviluppo del vaccino è una priorità dell’Organizzazione mondiale della sanità in quanto mancano farmaci efficaci per la gestione della malattia paramixovirale. Un disordine insolito nel complesso replicativo dei paramixovirus potrebbe rappresentare un potenziale bersaglio di nuove terapie, ma la caratterizzazione dei prerequisiti dei cambiamenti conformazionali dinamici nel meccanismo di replicazione è stata impegnativa. Il progetto DynamicAssemblies, finanziato dall’UE, prevede di chiarire quel meccanismo con una risoluzione atomica senza precedenti, il primo passo fondamentale per lo sviluppo mirato di farmaci.
Obiettivo
Paramyxoviruses, including, measles and a number of dangerous human pathogens, are negative strand RNA viruses that express their own machinery for transcription and replication. Different interactions between the nucleoprotein (N) and the phosphoprotein (P) are essential for chaperoning and assembly of N on newly synthesized RNA genomes to form nucleocapsids (NCs), as well as for initiating replication and transcription. Both N and tetrameric P exhibit extensive conformational disorder, with very long, unfolded regions that host important post-translational modification sites as well as regulatory interactions with host and viral partners. The presence of this level of disorder, in viruses whose genetic information is normally so parsimoniously exploited, remains unexplained. The elaboration of time-resolved, atomic resolution descriptions of the interaction trajectories of these highly disordered N:P complexes is extremely challenging, requiring the development of adapted methodologies that can account for their intrinsic flexibility. The role of N and P has been rendered yet more enigmatic following our recent observation that when mixed in solution they form liquid-like droplets. Such membraneless organelles are revolutionizing our understanding of cellular chemical biology, although their physical basis is poorly understood. Our aim is to describe these important complexes at atomic resolution, in particular to understand the role of the extensive conformational dynamics of N and P in the replication cycle. Our recent success in engineering soluble N:P complexes from measles that assemble into NCs, combined with ongoing development of NMR-based methods to investigate the structure, dynamics and interaction kinetics of large, intrinsically disordered proteins, fluorescence spectroscopy, cryoEM, SAXS, crystallography and molecular simulation, will provide the essential tools to investigate the functional mechanisms of these previously inaccessible complexes.
Campo scientifico
- medical and health scienceshealth sciencesinfectious diseasesRNA viruses
- natural sciencesbiological sciencesmicrobiologyvirology
- natural sciencesearth and related environmental sciencesgeologymineralogycrystallography
- natural sciencesphysical sciencesopticsspectroscopyemission spectroscopy
- natural sciencesbiological sciencesgeneticsRNA
Programma(i)
Argomento(i)
Meccanismo di finanziamento
ERC-ADG - Advanced GrantIstituzione ospitante
75015 PARIS 15
Francia