Archives of Virology

, Volume 158, Issue 6, pp 1151–1157 | Cite as

A serine-to-asparagine mutation at position 314 of H5N1 avian influenza virus NP is a temperature-sensitive mutation that interferes with nuclear localization of NP

  • Nattamon Siboonnan
  • Wittawat Wiriyarat
  • Chompunuch Boonarkart
  • Warunya Chakritbudsabong
  • Anan Jongkaewwattana
  • Pilaipan Puthavathana
  • Prasert Auewarakul
  • Ornpreya SuptawiwatEmail author
Original Article


We have generated a temperature-sensitive (ts) mutant from a human isolate of the H5N1 avian influenza virus by classical adaptation in cell culture. After 20 passages at low temperature, the virus showed a ts phenotype. The ts mutant also showed an attenuated phenotype after nasal inoculation in mice. Using reverse genetics, we generated reassortants carrying individual genomic segments of the wild-type and mutant viruses in an A/Puerto Rico/8/34 background, and found that the nucleoprotein (NP) gene could confer the ts phenotype. This mutant NP contains a serine-to-asparagine mutation at position 314 (S314N). The mutant NP protein showed a defect in nuclear localization at high temperature in mammalian cells.


Influenza Virus MDCK Cell Avian Influenza Virus H5N1 Avian Influenza Virus S314N Mutation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported by Thailand Research Fund (TRG5180011). S.N. and B.C. were supported by the postgraduate scholarship and research assistant programs, respectively, of the Faculty of Medicine Siriraj Hospital, pHw2000 plasmid was kindly provided by R.G. Webster.


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Copyright information

© Springer-Verlag Wien 2013

Authors and Affiliations

  • Nattamon Siboonnan
    • 1
  • Wittawat Wiriyarat
    • 2
  • Chompunuch Boonarkart
    • 1
  • Warunya Chakritbudsabong
    • 2
  • Anan Jongkaewwattana
    • 3
  • Pilaipan Puthavathana
    • 1
  • Prasert Auewarakul
    • 1
  • Ornpreya Suptawiwat
    • 1
    Email author
  1. 1.Department of Microbiology, Faculty of Medicine Siriraj HospitalMahidol UniversityBangkokThailand
  2. 2.Department of Preclinic and Applied Animal Science, Faculty of Veterinary ScienceMahidol UniversityNakhonpathomThailand
  3. 3.National Center of Genetic Engineering and BiotechnologyPathumthaniThailand

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