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Journal of Virology, March 2006, p. 2318-2325, Vol. 80, No. 5
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.5.2318-2325.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Hierarchy among Viral RNA (vRNA) Segments in Their Role in vRNA Incorporation into Influenza A Virions

Yukiko Muramoto,1,2,3 Ayato Takada,2,3 Ken Fujii,2,3 Takeshi Noda,3,4 Kiyoko Iwatsuki-Horimoto,2,3 Shinji Watanabe,5 Taisuke Horimoto,2,3 Hiroshi Kida,1 and Yoshihiro Kawaoka2,3,4,5*

Laboratory of Microbiology, Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo 060-0818,1 Division of Virology, Department of Microbiology and Immunology,2 International Research Center for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639,4 Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan,3 Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin—Madison, Madison, Wisconsin 537065

Received 29 August 2005/ Accepted 6 December 2005

The genome of influenza A viruses comprises eight negative-strand RNA segments. Although all eight segments must be present in cells for efficient viral replication, the mechanism(s) by which these viral RNA (vRNA) segments are incorporated into virions is not fully understood. We recently found that sequences at both ends of the coding regions of the HA, NA, and NS vRNA segments of A/WSN/33 play important roles in the incorporation of these vRNAs into virions. In order to similarly identify the regions of the PB2, PB1, and PA vRNAs of this strain that are critical for their incorporation, we generated a series of mutant vRNAs that possessed the green fluorescent protein gene flanked by portions of the coding and noncoding regions of the respective segments. For all three polymerase segments, deletions at the ends of their coding regions decreased their virion incorporation efficiencies. More importantly, these regions not only affected the incorporation of the segment in which they reside, but were also important for the incorporation of other segments. This effect was most prominent with the PB2 vRNA. These findings suggest a hierarchy among vRNA segments for virion incorporation and may imply intersegment association of vRNAs during virus assembly.

* Corresponding author. Mailing address: Institute of Medical Science, University of Tokyo, Shirokanedai, Minato-ku, Tokyo 108-8639, Japan. Phone: 81-3-5449-5310. Fax: 81-3-5449-5408. E-mail: kawaoka{at}ims.u-tokyo.ac.jp.

Journal of Virology, March 2006, p. 2318-2325, Vol. 80, No. 5
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.5.2318-2325.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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