J. Virol., Aug 1996, 5016-5024, Vol 70, No. 8
Copyright © 1996, American Society for Microbiology

Rescue of a synthetic chloramphenicol acetyltransferase RNA into influenza virus-like particles obtained from recombinant plasmids

I Mena, A Vivo, E Perez and A Portela
Centro Nacional de Biologia Celular y Retrovirus, Instituto de Salud Carlos III, Madrid, Spain.

We have shown previously that COS-1 cells infected with a vaccinia virus recombinant (vTF7-3) which expresses the T7 RNA polymerase gene and then transfected with four pGEM-derived plasmids encoding the influenza A virus core proteins (nucleoprotein, PB1, PB2, and PA polypeptides) can express a synthetic influenza virus-like chloramphenicol [correction of chloramphenical] acetyltransferase (CAT) RNA (I. Mena, S. de la Luna, C. Albo, J. Martin, A. Nieto, J. Ortin, and A. Portela, J. Gen. Virol. 75:2109-2114, 1994). Here we demonstrate that by supplying the vTF7-3-infected cells with plasmids containing cDNAs of all 10 influenza virus-encoded proteins, the transfected CAT RNA can be expressed and rescued into particles that are budded into the supernatant fluids. The released particles can transfer the enclosed CAT RNA to MDCK cultures and resemble true influenza virions in that they require trypsin treatment to deliver the RNA to fresh cells and are neutralized by a monoclonal antibody specific for the influenza A virus hemagglutinin. Moreover, analysis by electron microscopy showed that the culture medium harvested from the transfected cells contained vesicles that could be labeled with an anti- HA monoclonal antibody and that were similar in size and morphology to authentic influenza virus particles. It is also shown that detection of recombinant particles capable of transmitting the CAT RNA does not require expression of the influenza virus nonstructural protein NS1. All of these data indicate that influenza virus-like particles enclosing a synthetic virus-like RNA can be assembled in cells expressing all viral structural components from recombinant plasmids.

This article has been cited by other articles:

• Chen, B. J., Leser, G. P., Morita, E., Lamb, R. A. (2007). Influenza Virus Hemagglutinin and Neuraminidase, but Not the Matrix Protein, Are Required for Assembly and Budding of Plasmid-Derived Virus-Like Particles. J. Virol. 81: 7111-7123 [Abstract] [Full Text]  
• McCown, M. F., Pekosz, A. (2006). Distinct domains of the influenza a virus m2 protein cytoplasmic tail mediate binding to the m1 protein and facilitate infectious virus production.. J. Virol. 80: 8178-8189 [Abstract] [Full Text]  
• Cheung, T. K. W., Guan, Y., Ng, S. S. F., Chen, H., Wong, C. H. K., Peiris, J. S. M., Poon, L. L. M. (2005). Generation of recombinant influenza A virus without M2 ion-channel protein by introduction of a point mutation at the 5' end of the viral intron. J. Gen. Virol. 86: 1447-1454 [Abstract] [Full Text]  
• Muraki, Y., Washioka, H., Sugawara, K., Matsuzaki, Y., Takashita, E., Hongo, S. (2004). Identification of an amino acid residue on influenza C virus M1 protein responsible for formation of the cord-like structures of the virus. J. Gen. Virol. 85: 1885-1893 [Abstract] [Full Text]  
• Neumann, G., Whitt, M. A., Kawaoka, Y. (2002). A decade after the generation of a negative-sense RNA virus from cloned cDNA - what have we learned?. J. Gen. Virol. 83: 2635-2662 [Abstract] [Full Text]  
• Lommer, B. S., Luo, M. (2002). Structural Plasticity in Influenza Virus Protein NS2 (NEP). J. Biol. Chem. 277: 7108-7117 [Abstract] [Full Text]  
• Paragas, J., Talon, J., O'Neill, R. E., Anderson, D. K., Garcia-Sastre, A., Palese, P. (2001). Influenza B and C Virus NEP (NS2) Proteins Possess Nuclear Export Activities. J. Virol. 75: 7375-7383 [Abstract] [Full Text]  
• Bullido, R., Gómez-Puertas, P., Saiz, M. J., Portela, A. (2001). Influenza A Virus NEP (NS2 Protein) Downregulates RNA Synthesis of Model Template RNAs. J. Virol. 75: 4912-4917 [Abstract] [Full Text]  
• Crescenzo-Chaigne, B., van der Werf, S. (2001). Nucleotides at the extremities of the viral RNA of influenza C virus are involved in type-specific interactions with the polymerase complex. J. Gen. Virol. 82: 1075-1083 [Abstract] [Full Text]  
• Elton, D., Simpson-Holley, M., Archer, K., Medcalf, L., Hallam, R., McCauley, J., Digard, P. (2001). Interaction of the Influenza Virus Nucleoprotein with the Cellular CRM1-Mediated Nuclear Export Pathway. J. Virol. 75: 408-419 [Abstract] [Full Text]  
• Gómez-Puertas, P., Albo, C., Pérez-Pastrana, E., Vivo, A., Portela, A. (2000). Influenza Virus Matrix Protein Is the Major Driving Force in Virus Budding. J. Virol. 74: 11538-11547 [Abstract] [Full Text]  
• Wagner, E., Engelhardt, O. G., Weber, F., Haller, O., Kochs, G. (2000). Formation of virus-like particles from cloned cDNAs of Thogoto virus. J. Gen. Virol. 81: 2849-2853 [Abstract] [Full Text]  
• Lee, K. J., Novella, I. S., Teng, M. N., Oldstone, M. B. A., de la Torre, J. C. (2000). NP and L Proteins of Lymphocytic Choriomeningitis Virus (LCMV) Are Sufficient for Efficient Transcription and Replication of LCMV Genomic RNA Analogs. J. Virol. 74: 3470-3477 [Abstract] [Full Text]  
• Neumann, G., Watanabe, T., Kawaoka, Y. (2000). Plasmid-Driven Formation of Influenza Virus-Like Particles. J. Virol. 74: 547-551 [Abstract] [Full Text]  
• Neumann, G., Watanabe, T., Ito, H., Watanabe, S., Goto, H., Gao, P., Hughes, M., Perez, D. R., Donis, R., Hoffmann, E., Hobom, G., Kawaoka, Y. (1999). Generation of influenza A viruses entirely from cloned cDNAs. Proc. Natl. Acad. Sci. USA 96: 9345-9350 [Abstract] [Full Text]  
• Teng, M. N., Collins, P. L. (1998). Identification of the Respiratory Syncytial Virus Proteins Required for Formation and Passage of Helper-Dependent Infectious Particles. J. Virol. 72: 5707-5716 [Abstract] [Full Text]  
• Hoffmann, E., Neumann, G., Kawaoka, Y., Hobom, G., Webster, R. G. (2000). A DNA transfection system for generation of influenza A virus from eight plasmids. Proc. Natl. Acad. Sci. USA 97: 6108-6113 [Abstract] [Full Text]