A genome-wide analysis of the spatio-temporal distribution of herpesviral proteins in the course of infection

• Antragstellerin: Privatdozentin Dr. Susanne Bailer, Ph.D.
• Fachliche Zuordnung: Virologie
• Förderung: Förderung von 2008 bis 2013
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 52364154

Crucial aspects of the herpesviral life cycle including DNA replication, gene transcription and RNA export, capsid morphogenesis, and egress occur in the host nucleus and require that numerous herpesviral proteins are transported between the host cytosol and the nucleus. The coordinated spatio-temporal distribution of viral proteins between nucleus and cytoplasm is thus vital for viral replication. The docking of incoming herpesviral capsids to the nuclear pore complex (NPC) and the release of viral DNA into the nuclear interior represents a variant of nuclear import and another critical step of viral infection. In general, transport of proteins and some RNAs between cytoplasm and nucleus is mediated by a direct or indirect binding to transport factors of the importin ( and (-families and occurs along a gradient of the small GTPase Ran. So far the mechanism of nucleo-cytoplasmic transport is unknown for a large number of viral proteins and those viral proteins mediating capsid docking have not been identified. In this collaborative project between two groups with high expertise in nucleocytoplasmic transport (S. Bailer) and high-throughput-based screening technologies in infectious diseases (J. Haas), we will apply a novel approach to comprehensively analyse the interactions between herpesviral proteins and all known transport factors of the eukaryotic importin ( and (-families by performing large-scale yeast-two-hybrid (Y2H) screens and in vitro biochemical assays in parallel. The novel viral cargoes of these transport factors will be functionally verified and the subcellular localization of viral proteins and their nuclear transport signals determined systematically. The data received will then be combined by bioinformatical tools to create a dynamic model of the spatio-temporal distribution of viral proteins and capsids during herpesviral infection. This analysis will not only considerably increase our understanding of herpesviral biology and reveal potential novel targets for viral treatment, but also improve the in silico prediction of novel consensus sequences for cellular nuclear transport cargoes.

• DFG-Verfahren: Sachbeihilfen
• Beteiligte Person: Professor Dr. Jürgen Haas