2 Aim of the Thesis

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In anticipation of a pandemic outbreak of influenza A virus with respect to the persistent threat of evolving human and avian variants with highly pathogenic potential detailed knowledge about the viral replication cycle and impact on the host cell are urgently ask for. Influenza A virus hijacks cellular biosynthetic pathways, intracellular transport mechanisms and biomolecular resources while interacting with host proteins for an efficient replication and production of progeny virus particles. Every single step of the replication cycle from entry to budding needs to be investigated to allow the development of preventive and therapeutic antiviral strategies.

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Therefore this work focuses on the viral transcription and translation as well as the involvement and response of the host cell.

Due to essential drawbacks of existing nucleic acid detection techniques in the cellular context the development of a widely applicable, highly specific and sensitive method to study viral mRNA is one of the most relevant issues. The present thesis was undertaken to explore the suitability of FIT-PNA molecules to investigate the viral transcription in the early phase of influenza A virus infection in living host cells. To assess the spatial, temporal and quantitative progression of viral mRNA molecules a specific RT-qPCR shall be combined with a study in living infected cells using CLSM.

Previous studies already assessed the alterations of the cellular proteome after infection to identify viral host-interaction partners. With the help of quantitative proteomics based on mass spectrometry the early phase of an influenza A virus infection shall be characterized to reveal host factors involved in viral replication and analyse the temporal progression of viral protein synthesis.


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