title: Experimental Approaches to Study Genome Packaging of Influenza A Viruses.
abstract: The genome of influenza A viruses (IAV) consists of eight single-stranded negative sense viral RNAs (vRNAs) encapsidated into viral ribonucleoproteins (vRNPs). It is now well established that genome packaging (i.e., the incorporation of a set of eight distinct vRNPs into budding viral particles), follows a specific pathway guided by segment-specific cis-acting packaging signals on each vRNA. However, the precise nature and function of the packaging signals, and the mechanisms underlying the assembly of vRNPs into sub-bundles in the cytoplasm and their selective packaging at the viral budding site, remain largely unknown. Here, we review the diverse and complementary methods currently being used to elucidate these aspects of the viral cycle. They range from conventional and competitive reverse genetics, single molecule imaging of vRNPs by fluorescence in situ hybridization (FISH) and high-resolution electron microscopy and tomography of budding viral particles, to solely in vitro approaches to investigate vRNA-vRNA interactions at the molecular level.
authors: Architecture et Réactivité de l'ARN, Université de Strasbourg, Centre National de la Recherche Scientifique (CNRS), Institut de Biologie Moléculaire et Cellulaire (IBMC), 15 rue René Descartes, 67084 Strasbourg, France. c.isel@ibmc-cnrs.unistra.fr.|Département de Virologie, Unité de Génétique Moléculaire des Virus à ARN, Institut Pasteur, 75015 Paris, France. c.isel@ibmc-cnrs.unistra.fr.
authors: Catherine
authors: C
authors: Isel
authors: Département de Virologie, Unité de Génétique Moléculaire des Virus à ARN, Institut Pasteur, 75015 Paris, France. sandie.munier@pasteur.fr.|Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 3569, 75016 Paris, France. sandie.munier@pasteur.fr.|Unité de Génétique Moléculaire des Virus à ARN, Sorbonne Paris Cité, Université Paris Diderot, 75013 Paris, France. sandie.munier@pasteur.fr.
authors: Sandie
authors: S
authors: Munier
authors: Département de Virologie, Unité de Génétique Moléculaire des Virus à ARN, Institut Pasteur, 75015 Paris, France. nadia.naffakh@pasteur.fr.|Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 3569, 75016 Paris, France. nadia.naffakh@pasteur.fr.|Unité de Génétique Moléculaire des Virus à ARN, Sorbonne Paris Cité, Université Paris Diderot, 75013 Paris, France. nadia.naffakh@pasteur.fr.
authors: Nadia
authors: N
authors: Naffakh
mesh_terms: D055032:Electron Microscope Tomography; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D009980:Influenza A virus / Q000502:physiology*; D046529:Microscopy, Electron, Transmission; D059386:Reverse Genetics; D000072760:Single Molecule Imaging; D014773:Virology / Q000379:methods*; D019065:Virus Assembly
keywords: RNA-RNA interaction; competitive reverse genetics; influenza virus; packaging assay; packaging signal; single-molecule FISH