This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Biswas, S. K.
Right arrow Articles by Nayak, D. P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Biswas, S. K.
Right arrow Articles by Nayak, D. P.

 Previous Article  |  Next Article 

J Virol, July 1998, p. 5493-5501, Vol. 72, No. 7
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Influenza Virus Nucleoprotein Interacts with Influenza Virus Polymerase Proteins

Siddhartha K. Biswas, Paul L. Boutz, and Debi P. Nayak*

Department of Microbiology and Immunology, Jonsson Comprehensive Cancer Center, UCLA School of Medicine, Los Angeles, California 90095-1747

Received 29 January 1998/Accepted 30 March 1998

Influenza virus nucleoprotein (NP) is a critical factor in the viral infectious cycle in switching influenza virus RNA synthesis from transcription mode to replication mode. In this study, we investigated the interaction of NP with the viral polymerase protein complex. Using coimmunoprecipitation with monospecific or monoclonal antibodies, we observed that NP interacted with the RNP-free polymerase protein complex in influenza virus-infected cells. In addition, coexpression of the components of the polymerase protein complex (PB1, PB2, or PA) with NP either together or pairwise revealed that NP interacts with PB1 and PB2 but not PA. Interaction of NP with PB1 and PB2 was confirmed by both coimmunoprecipitation and histidine tagging of the NP-PB1 and NP-PB2 complexes. Further, it was observed that NP-PB2 interaction was rather labile and sensitive to dissociation in 0.1% sodium dodecyl sulfate and that the stability of NP-PB2 interaction was regulated by the sequences present at the COOH terminus of NP. Analysis of NP deletion mutants revealed that at least three regions of NP interacted independently with PB2. A detailed analysis of the COOH terminus of NP by mutation of serine-to-alanine (SA) residues either individually or together demonstrated that SA mutations in this region did not affect the binding of NP to PB2. However, some SA mutations at the COOH terminus drastically affected the functional activity of NP in an in vivo transcription-replication assay, whereas others exhibited a temperature-sensitive phenotype and still others had no effect on the transcription and replication of the viral RNA. These results suggest that a direct interaction of NP with polymerase proteins may be involved in regulating the switch of viral RNA synthesis from transcription to replication.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Jonsson Comprehensive Cancer Center, UCLA School of Medicine, 10833 Le Conte Ave., Los Angeles, CA 90095-1747. Phone: (310) 825-8558. Fax: (310) 206-3865. E-mail: dnayak{at}ucla.edu.

J Virol, July 1998, p. 5493-5501, Vol. 72, No. 7
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Li, Z., Watanabe, T., Hatta, M., Watanabe, S., Nanbo, A., Ozawa, M., Kakugawa, S., Shimojima, M., Yamada, S., Neumann, G., Kawaoka, Y. (2009). Mutational Analysis of Conserved Amino Acids in the Influenza A Virus Nucleoprotein. J. Virol. 83: 4153-4162 [Abstract] [Full Text]  
  • Noton, S. L., Simpson-Holley, M., Medcalf, E., Wise, H. M., Hutchinson, E. C., McCauley, J. W., Digard, P. (2009). Studies of an Influenza A Virus Temperature-Sensitive Mutant Identify a Late Role for NP in the Formation of Infectious Virions. J. Virol. 83: 562-571 [Abstract] [Full Text]  
  • Newcomb, L. L., Kuo, R.-L., Ye, Q., Jiang, Y., Tao, Y. J., Krug, R. M. (2009). Interaction of the Influenza A Virus Nucleocapsid Protein with the Viral RNA Polymerase Potentiates Unprimed Viral RNA Replication. J. Virol. 83: 29-36 [Abstract] [Full Text]  
  • Ng, A. K.-L., Zhang, H., Tan, K., Li, Z., Liu, J.-h., Chan, P. K.-S., Li, S.-M., Chan, W.-Y., Au, S. W.-N., Joachimiak, A., Walz, T., Wang, J.-H., Shaw, P.-C. (2008). Structure of the influenza virus A H5N1 nucleoprotein: implications for RNA binding, oligomerization, and vaccine design. FASEB J. 22: 3638-3647 [Abstract] [Full Text]  
  • Wasilenko, J. L., Lee, C. W., Sarmento, L., Spackman, E., Kapczynski, D. R., Suarez, D. L., Pantin-Jackwood, M. J. (2008). NP, PB1, and PB2 Viral Genes Contribute to Altered Replication of H5N1 Avian Influenza Viruses in Chickens. J. Virol. 82: 4544-4553 [Abstract] [Full Text]  
  • Tsfasman, T. M., Markushin, S. G., Akopova, I. I., Ghendon, Y. Z. (2007). Molecular mechanisms of reversion to the ts+ (non-temperature-sensitive) phenotype of influenza A cold-adapted (ca) virus strains. J. Gen. Virol. 88: 2724-2729 [Abstract] [Full Text]  
  • Torreira, E., Schoehn, G., Fernandez, Y., Jorba, N., Ruigrok, R. W.H., Cusack, S., Ortin, J., Llorca, O. (2007). Three-dimensional model for the isolated recombinant influenza virus polymerase heterotrimer. Nucleic Acids Res 35: 3774-3783 [Abstract] [Full Text]  
  • Vreede, F. T., Brownlee, G. G. (2007). Influenza Virion-Derived Viral Ribonucleoproteins Synthesize both mRNA and cRNA In Vitro. J. Virol. 81: 2196-2204 [Abstract] [Full Text]  
  • Mullin, A. E., Dalton, R. M., Amorim, M. J., Elton, D., Digard, P. (2004). Increased amounts of the influenza virus nucleoprotein do not promote higher levels of viral genome replication. J. Gen. Virol. 85: 3689-3698 [Abstract] [Full Text]  
  • Fodor, E., Smith, M. (2004). The PA Subunit Is Required for Efficient Nuclear Accumulation of the PB1 Subunit of the Influenza A Virus RNA Polymerase Complex. J. Virol. 78: 9144-9153 [Abstract] [Full Text]  
  • Vreede, F. T., Jung, T. E., Brownlee, G. G. (2004). Model Suggesting that Replication of Influenza Virus Is Regulated by Stabilization of Replicative Intermediates. J. Virol. 78: 9568-9572 [Abstract] [Full Text]  
  • Area, E., Martin-Benito, J., Gastaminza, P., Torreira, E., Valpuesta, J. M., Carrascosa, J. L., Ortin, J. (2004). 3D structure of the influenza virus polymerase complex: Localization of subunit domains. Proc. Natl. Acad. Sci. USA 101: 308-313 [Abstract] [Full Text]  
  • Gastaminza, P., Perales, B., Falcon, A. M., Ortin, J. (2003). Mutations in the N-Terminal Region of Influenza Virus PB2 Protein Affect Virus RNA Replication but Not Transcription. J. Virol. 77: 5098-5108 [Abstract] [Full Text]  
  • Portela, A., Digard, P. (2002). The influenza virus nucleoprotein: a multifunctional RNA-binding protein pivotal to virus replication. J. Gen. Virol. 83: 723-734 [Abstract] [Full Text]  
  • Lee, M. T. M., Bishop, K., Medcalf, L., Elton, D., Digard, P., Tiley, L. (2002). Definition of the minimal viral components required for the initiation of unprimed RNA synthesis by influenza virus RNA polymerase. Nucleic Acids Res 30: 429-438 [Abstract] [Full Text]  
  • Perez, D. R., Donis, R. O. (2001). Functional Analysis of PA Binding by Influenza A Virus PB1: Effects on Polymerase Activity and Viral Infectivity. J. Virol. 75: 8127-8136 [Abstract] [Full Text]  
  • Naffakh, N., Massin, P., Escriou, N., Crescenzo-Chaigne, B., van der Werf, S. (2000). Genetic analysis of the compatibility between polymerase proteins from human and avian strains of influenza A viruses. J. Gen. Virol. 81: 1283-1291 [Abstract] [Full Text]  
  • Perales, B., Sanz-Ezquerro, J. J., Gastaminza, P., Ortega, J., Santarén, J. F., Ortín, J., Nieto, A. (2000). The Replication Activity of Influenza Virus Polymerase Is Linked to the Capacity of the PA Subunit To Induce Proteolysis. J. Virol. 74: 1307-1312 [Abstract] [Full Text]  
  • Medcalf, L., Poole, E., Elton, D., Digard, P. (1999). Temperature-Sensitive Lesions in Two Influenza A Viruses Defective for Replicative Transcription Disrupt RNA Binding by the Nucleoprotein. J. Virol. 73: 7349-7356 [Abstract] [Full Text]  
  • Elton, D., Medcalf, L., Bishop, K., Harrison, D., Digard, P. (1999). Identification of Amino Acid Residues of Influenza Virus Nucleoprotein Essential for RNA Binding. J. Virol. 73: 7357-7367 [Abstract] [Full Text]  
  • Mena, I., Jambrina, E., Albo, C., Perales, B., Ortín, J., Arrese, M., Vallejo, D., Portela, A. (1999). Mutational Analysis of Influenza A Virus Nucleoprotein: Identification of Mutations That Affect RNA Replication. J. Virol. 73: 1186-1194 [Abstract] [Full Text]  
  • Uhrig, J. F., Soellick, T.-R., Minke, C. J., Philipp, C., Kellmann, J.-W., Schreier, P. H. (1999). Homotypic interaction and multimerization of nucleocapsid protein of tomato spotted wilt tospovirus: Identification and characterization of two interacting domains. Proc. Natl. Acad. Sci. USA 96: 55-60 [Abstract] [Full Text]  
  • Majumder, A., Basak, S., Raha, T., Chowdhury, S. P., Chattopadhyay, D., Roy, S. (2001). Effect of Osmolytes and Chaperone-like Action of P-protein on Folding of Nucleocapsid Protein of Chandipura Virus. J. Biol. Chem. 276: 30948-30955 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»