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The role of the N-terminal caspase cleavage site in the nucleoprotein of influenza A virus in vitro and in vivo

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Abstract

The N-terminal caspase cleavage in the nucleoprotein (NP) of influenza A virus is correlated with the host origin of the virus, thus could be a molecular determinant for host range. We studied how mutations targeting the NP cleavage motif of human and avian influenza viruses affect virus replication in vitro and in vivo. The “avian-like” D16→G substitution in the NP, which makes this protein resistant to cleavage, did not significantly affect the human A/Puerto Rico/8/34 (H1N1) virus replication in vitro but decreased the lethality of this virus in mice by 68-fold. Gene incompatibility contributed to the attenuated phenotype of the reassortant A/Puerto Rico/8/34 virus with avian NP derived from A/Teal/Hong Kong/w312/97 (H6N1) virus in vitro and in vivo. Insertion of the “human-like” G16→D mutation into avian NP, which resulted in susceptibility to caspase cleavage, did not rescue virulence, but made the reassortant virus even more attenuated. Introducing the human-like G16→D substitution into the NP of highly pathogenic A/Vietnam/1203/04 (H5N1) virus decreased lethality in mice. We confirmed that position 16, which associated with the N-terminal caspase cleavage of the NP, is important for optimal virus fitness in vitro and in vivo. An avian-like mutation at position 16 in the NP of human virus as well as a human-like substitution at this residue in avian NP both resulted in virus attenuation.

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Acknowledgments

We thank Jon P. Seiler for excellent technical assistance. Special thanks to Julia Cay Jones for editing the manuscript and Kristina S. Makarova for illustrations. This study was supported by NIAID contract AI-95357, by NIH Cancer Center Support (CORE) grant CA-21765, and by the American Lebanese Syrian Associated Charities (ALSAC).

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Author notes

  1. H. Ozaki

    Present address: Division of Project Research, Creative Research Initiative, Hokkaido University, Kita 21, Nishi 10, Kita-ku, Sapporo, 001-0021, Japan

Authors and Affiliations

  1. Division of Virology, Department of Infectious Diseases, St Jude Children’s Research Hospital, 332 North Lauderdale Street, Memphis, TN, 38105, USA

    A. S. Lipatov, H.-L. Yen, R. Salomon, H. Ozaki, E. Hoffmann & R. G. Webster

  2. United States Department of Agriculture, Southeast Poultry Research Laboratory, Agricultural Research Service, 934 College Station Road, Athens, GA, 30605, USA

    A. S. Lipatov

Authors
  1. A. S. Lipatov
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  2. H.-L. Yen
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  3. R. Salomon
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  4. H. Ozaki
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  5. E. Hoffmann
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  6. R. G. Webster
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Corresponding author

Correspondence to R. G. Webster.

Additional information

These data have been presented in part at The 3rd Orthomyxovirus Research Conference, July 28–31, 2005. Queen’s College, Cambridge, UK.

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Lipatov, A.S., Yen, HL., Salomon, R. et al. The role of the N-terminal caspase cleavage site in the nucleoprotein of influenza A virus in vitro and in vivo. Arch Virol 153, 427–434 (2008). https://doi.org/10.1007/s00705-007-0003-8

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  • DOI: https://doi.org/10.1007/s00705-007-0003-8

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