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Journal of Virology, April 2004, p. 3880-3888, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.3880-3888.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Defective RNA Replication and Late Gene Expression in Temperature-Sensitive Influenza Viruses Expressing Deleted Forms of the NS1 Protein

Ana M. Falcón,1 Rosa M. Marión,1 Thomas Zürcher,1,{dagger} Paulino Gómez,2,{ddagger} Agustín Portela,2 Amelia Nieto,1 and Juan Ortín1*

Centro Nacional de Biotecnología, CSIC, 28049 Madrid,1 División de Productos Biológicos y Biotecnología, Agencia Española del Medicamento, 28220 Madrid, Spain2

Received 3 September 2003/ Accepted 12 December 2003

Influenza A virus mutants expressing C-terminally deleted forms of the NS1 protein (NS1-81 and NS1-110) were generated by plasmid rescue. These viruses were temperature sensitive and showed a small plaque size at the permissive temperature. The accumulation of virion RNA in mutant virus-infected cells was reduced at the restrictive temperature, while the accumulation of cRNA or mRNA was not affected, indicating that the NS1 protein is involved in the control of transcription versus replication processes in the infection. The synthesis and accumulation of late virus proteins were reduced in NS1-81 mutant-infected cells at the permissive temperature and were essentially abolished for both viruses at the restrictive temperature, while synthesis and accumulation of nucleoprotein (NP) were unaffected. Probably as a consequence, the nucleocytoplasmic export of virus NP was strongly inhibited at the restrictive temperature. These results indicate that the NS1 protein is essential for nuclear and cytoplasmic steps during the virus cycle.

* Corresponding author. Mailing address: Centro Nacional de Biotecnología, CSIC, Campus de Cantoblanco, 28049 Madrid, Spain. Phone: 34 91 585 4557. Fax: 34 91 585 4506. E-mail: jortin{at}cnb.uam.es.

{dagger} Present address: GlaxoSmithKline Medicines Research Centre, Stevenage, Herts SG1 2NY, United Kingdom.

{ddagger} Present address: Centro de Astrobiología (CAB-CSIC), 28550 Madrid, Spain.

Journal of Virology, April 2004, p. 3880-3888, Vol. 78, No. 8
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.8.3880-3888.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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