Descripción del proyecto
Descubrir el orden oculto en el aparente desorden puede estimular el desarrollo de fármacos específicos
Muchas dolencias respiratorias y varias enfermedades prevalentes en la infancia como el sarampión y las paperas se deben a paramixovirus. En Estados Unidos, el virus sincitial respiratorio (VSR) es la principal causa de hospitalización infantil. El desarrollo de vacunas es una prioridad de la Organización Mundial de la Salud, ya que se carece de medicamentos eficaces para controlar las enfermedades paramixovirales. Un trastorno poco común en el complejo replicativo de los paramixovirus podría ser una posible diana para nuevas terapias, pero la caracterización previa de los cambios conformacionales en la maquinaria de replicación está resultando complicada. El proyecto financiado con fondos europeos DynamicAssemblies prevé elucidar esa maquinaria con una resolución atómica sin precedentes; un primer paso fundamental hacia el desarrollo de fármacos específicos.
Objetivo
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.
Ámbito científico
- natural sciencesbiological sciencesmicrobiologyvirology
- natural sciencesearth and related environmental sciencesgeologymineralogycrystallography
- natural sciencesphysical sciencesopticsspectroscopyemission spectroscopy
- natural sciencesbiological sciencesgeneticsRNA
- natural sciencesbiological sciencesgeneticsgenomes
Programa(s)
Régimen de financiación
ERC-ADG - Advanced GrantInstitución de acogida
75015 PARIS 15
Francia