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Viruses 2016, 8(8), 218; doi:10.3390/v8080218

Experimental Approaches to Study Genome Packaging of Influenza A Viruses

1,2,* , 2,3,4
and
2,3,4,*
1
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
2
Département de Virologie, Unité de Génétique Moléculaire des Virus à ARN, Institut Pasteur, 75015 Paris, France
3
Centre National de la Recherche Scientifique (CNRS), Unité Mixte de Recherche 3569, 75016 Paris, France
4
Unité de Génétique Moléculaire des Virus à ARN, Sorbonne Paris Cité, Université Paris Diderot, 75013 Paris, France
*
Authors to whom correspondence should be addressed.
Academic Editor: Polly Roy
Received: 10 June 2016 / Revised: 26 July 2016 / Accepted: 1 August 2016 / Published: 9 August 2016
(This article belongs to the Special Issue RNA Packaging)
View Full-Text   |   Download PDF [2437 KB, uploaded 9 August 2016]   |  

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. View Full-Text
Keywords: influenza virus; packaging signal; packaging assay; single-molecule FISH; RNA-RNA interaction; competitive reverse genetics influenza virus; packaging signal; packaging assay; single-molecule FISH; RNA-RNA interaction; competitive reverse genetics
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Isel, C.; Munier, S.; Naffakh, N. Experimental Approaches to Study Genome Packaging of Influenza A Viruses. Viruses 2016, 8, 218.

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