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The Incorporation of Influenza A Virus Genome Segments Does Not Absolutely Require Wild-Type Sequences

¹ Institute of Medical Science, University of Tokyo;
² University of Wisconsin-Madison

³ E-mail: kawaokay{at}svm.vetmed.wisc.edu

The efficient incorporation of influenza virus genome segments into virions is mediated by cis-acting regions at both ends of the viral RNAs. We previously showed that nucleotides 16 to 26 at the 3' end of the NS viral RNA are important for efficient virion incorporation and that nucleotides 27 to 56 also contribute to this process. To further understand the signaling requirements for genome packaging, we performed linker-scanning mutagenesis in this latter region and found that nucleotides 27 to 35 make an appreciable contribution to the efficient incorporation of the NS segment. We then generated an NS vRNA library composed of an RNA population with randomized nucleotides at positions 16 to 35 such that the virus could select the sequences it required for virion incorporation. The sequences selected differed from the wild-type sequence and no conserved nucleotides were selected. The ability of non-wild-type sequences to function in this manner indicates that the incorporation of influenza A virus genome segments does not absolutely require specific sequences.

Received 16 January 2009; accepted 18 March 2009.