Abstract
The influenza A virus genome consists of eight-segmented, single-stranded, negative-sense RNAs. Each genomic viral RNA segment (vRNA) encodes different viral proteins that are necessary for efficient virus replication, and forms a ribonucleoprotein complex (RNP) together with viral nucleoproteins and an RNA polymerase complex. Later in infection, progeny virions, which are released from the plasma membrane of the infected cell, must incorporate the eight separate vRNAs to be infectious. However, the mechanism by which the segmented vRNAs are incorporated into each progeny virion remains unclear. To elucidate the genome packaging mechanism of influenza A virus, we examined the architecture of RNPs within progeny virions using several electron microscopic analyses. We demonstrated that each progeny virion incorporates eight RNPs arranged in a specific pattern, in which seven RNPs surround the central one. Such characteristic arrangement is found in all influenza A virus strains tested here, suggesting that the mechanism by which well-organized eight RNPs is incorporated into virion is common to influenza A viruses. In addition, there seem to be physical interactions among the eight RNPs via nucleic acid-like structures, suggesting that there are specific interactions among the eight vRNAs in the form of RNPs. These results indicate that influenza A virion selectively packages a complete set of eight separate vRNAs.
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