Generation and Comprehensive Analysis of an Influenza Virus Polymerase Cellular Interaction Network

RNA Metabolism, IBMM, ULB, rue Profs. Jeener & Brachet 12, 6041 Gosselies, Belgium.
Journal of Virology (Impact Factor: 4.44). 12/2011; 85(24):13010-8. DOI: 10.1128/JVI.02651-10
Source: PubMed


The influenza virus transcribes and replicates its genome inside the nucleus of infected cells. Both activities are performed by the viral RNA-dependent RNA polymerase that is composed of the three subunits PA, PB1, and PB2, and recent studies have shown that it requires host cell factors to transcribe and replicate the viral genome. To identify these cellular partners, we generated a comprehensive physical interaction map between each polymerase subunit and the host cellular proteome. A total of 109 human interactors were identified by yeast two-hybrid screens, whereas 90 were retrieved by literature mining. We built the FluPol interactome network composed of the influenza virus polymerase (PA, PB1, and PB2) and the nucleoprotein NP and 234 human proteins that are connected through 279 viral-cellular protein interactions. Analysis of this interactome map revealed enriched cellular functions associated with the influenza virus polymerase, including host factors involved in RNA polymerase II-dependent transcription and mRNA processing. We confirmed that eight influenza virus polymerase-interacting proteins are required for virus replication and transcriptional activity of the viral polymerase. These are involved in cellular transcription (C14orf166, COPS5, MNAT1, NMI, and POLR2A), translation (EIF3S6IP), nuclear transport (NUP54), and DNA repair (FANCG). Conversely, we identified PRKRA, which acts as an inhibitor of the viral polymerase transcriptional activity and thus is required for the cellular antiviral response.

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Available from: Lionel Tafforeau,
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    • "When RNPs reach the nuclear membrane, nuclear importation is mediated by the binding of the nucleoprotein with the alpha importins, which then bind importin-β [129]. Simultaneously, the importins must interact with PA, PB1 and PB2 [130], and this affects the interaction of the RNP with the same importins [131]. Specifically, subunits PB1 and PA are imported by Ran- BP5 or karyopherin beta3 (also known as importin beta3, or importin 5), whilst subunit PB2 is imported by importin alpha-3 or importin alpha-7. "
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    • "Another strategy to identify host factors interacting with viral proteins is to use the yeast two-hybrid system (Shapira et al., 2009; Tafforeau et al., 2011). In particular, Tafforeau et al. (2011) used this system to identify several partners of H5N1 polymerase subunits. However, the approach is limited by a general high rate of false positives and the caveat of protein interactions not taking place in a relevant cellular context. "
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    • "In a separate recent screen, host genes specifically important for influenza viral polymerase (RdRP) function were identified, which are also potential candidates for antiviral drug development [136]. A chemical biology screen of 200,000 synthetic compounds recently identified naphthalimides as an antiviral chemical class that activated a new host defense factor, REDD1, which in turn inhibited influenza NS1 and viral replication [137]. "
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