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 Narayanan, K.
Right arrow Articles by Makino, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Narayanan, K.
Right arrow Articles by Makino, S.

 Previous Article  |  Next Article 

Journal of Virology, September 2000, p. 8127-8134, Vol. 74, No. 17
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of the Coronavirus M Protein and Nucleocapsid Interaction in Infected Cells

Krishna Narayanan, Akihiko Maeda, Junko Maeda, and Shinji Makino*

Department of Microbiology and Immunology, The University of Texas Medical Branch at Galveston, Galveston, Texas 77555-1019, and Department of Microbiology and Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, Texas 78712-1095

Received 30 March 2000/Accepted 8 June 2000

Coronavirus contains three envelope proteins, M, E and S, and a nucleocapsid, which consists of genomic RNA and N protein, within the viral envelope. We studied the macromolecular interactions involved in coronavirus assembly in cells infected with a murine coronavirus, mouse hepatitis virus (MHV). Coimmunoprecipitation analyses demonstrated an interaction between N protein and M protein in infected cells. Pulse-labeling experiments showed that newly synthesized, unglycosylated M protein interacted with N protein in a pre-Golgi compartment, which is part of the MHV budding site. Coimmunoprecipitation analyses further revealed that M protein interacted with only genomic-length MHV mRNA, mRNA 1, while N protein interacted with all MHV mRNAs. These data indicated that M protein interacted with the nucleocapsid, consisting of N protein and mRNA 1, in infected cells. The M protein-nucleocapsid interaction occurred in the absence of S and E proteins. Intracellular M protein-N protein interaction was maintained after removal of viral RNAs by RNase treatment. However, the M protein-N protein interaction did not occur in cells coexpressing M protein and N protein alone. These data indicated that while the M protein-N protein interaction, which is independent of viral RNA, occurred in the M protein-nucleocapsid complex, some MHV function(s) was necessary for the initiation of M protein-nucleocapsid interaction. The M protein-nucleocapsid interaction, which occurred near or at the MHV budding site, most probably represented the process of specific packaging of the MHV genome into MHV particles.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, The University of Texas Medical Branch at Galveston, Galveston, TX 77555-1019. Phone: (409) 772-2323. Fax: (409) 772-5065. E-mail: shmakino{at}utmb.edu.

Journal of Virology, September 2000, p. 8127-8134, Vol. 74, No. 17
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Siu, Y. L., Teoh, K. T., Lo, J., Chan, C. M., Kien, F., Escriou, N., Tsao, S. W., Nicholls, J. M., Altmeyer, R., Peiris, J. S. M., Bruzzone, R., Nal, B. (2008). The M, E, and N Structural Proteins of the Severe Acute Respiratory Syndrome Coronavirus Are Required for Efficient Assembly, Trafficking, and Release of Virus-Like Particles. J. Virol. 82: 11318-11330 [Abstract] [Full Text]  
  • Verma, S., Lopez, L. A., Bednar, V., Hogue, B. G. (2007). Importance of the Penultimate Positive Charge in Mouse Hepatitis Coronavirus A59 Membrane Protein. J. Virol. 81: 5339-5348 [Abstract] [Full Text]  
  • Huang, C., Peters, C. J., Makino, S. (2007). Severe Acute Respiratory Syndrome Coronavirus Accessory Protein 6 Is a Virion-Associated Protein and Is Released from 6 Protein-Expressing Cells. J. Virol. 81: 5423-5426 [Abstract] [Full Text]  
  • Saikatendu, K. S., Joseph, J. S., Subramanian, V., Neuman, B. W., Buchmeier, M. J., Stevens, R. C., Kuhn, P. (2007). Ribonucleocapsid Formation of Severe Acute Respiratory Syndrome Coronavirus through Molecular Action of the N-Terminal Domain of N Protein. J. Virol. 81: 3913-3921 [Abstract] [Full Text]  
  • Schaecher, S. R., Mackenzie, J. M., Pekosz, A. (2007). The ORF7b Protein of Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) Is Expressed in Virus-Infected Cells and Incorporated into SARS-CoV Particles. J. Virol. 81: 718-731 [Abstract] [Full Text]  
  • Huang, C., Ito, N., Tseng, C.-T. K., Makino, S. (2006). Severe acute respiratory syndrome coronavirus 7a accessory protein is a viral structural protein.. J. Virol. 80: 7287-7294 [Abstract] [Full Text]  
  • Neuman, B. W., Adair, B. D., Yoshioka, C., Quispe, J. D., Orca, G., Kuhn, P., Milligan, R. A., Yeager, M., Buchmeier, M. J. (2006). Supramolecular architecture of severe acute respiratory syndrome coronavirus revealed by electron cryomicroscopy.. J. Virol. 80: 7918-7928 [Abstract] [Full Text]  
  • Verma, S., Bednar, V., Blount, A., Hogue, B. G. (2006). Identification of Functionally Important Negatively Charged Residues in the Carboxy End of Mouse Hepatitis Coronavirus A59 Nucleocapsid Protein. J. Virol. 80: 4344-4355 [Abstract] [Full Text]  
  • Huang, C., Narayanan, K., Ito, N., Peters, C. J., Makino, S. (2006). Severe Acute Respiratory Syndrome Coronavirus 3a Protein Is Released in Membranous Structures from 3a Protein-Expressing Cells and Infected Cells. J. Virol. 80: 210-217 [Abstract] [Full Text]  
  • Hsieh, P.-K., Chang, S. C., Huang, C.-C., Lee, T.-T., Hsiao, C.-W., Kou, Y.-H., Chen, I-Y., Chang, C.-K., Huang, T.-H., Chang, M.-F. (2005). Assembly of Severe Acute Respiratory Syndrome Coronavirus RNA Packaging Signal into Virus-Like Particles Is Nucleocapsid Dependent. J. Virol. 79: 13848-13855 [Abstract] [Full Text]  
  • Youn, S., Collisson, E. W., Machamer, C. E. (2005). Contribution of Trafficking Signals in the Cytoplasmic Tail of the Infectious Bronchitis Virus Spike Protein to Virus Infection. J. Virol. 79: 13209-13217 [Abstract] [Full Text]  
  • Hurst, K. R., Kuo, L., Koetzner, C. A., Ye, R., Hsue, B., Masters, P. S. (2005). A Major Determinant for Membrane Protein Interaction Localizes to the Carboxy-Terminal Domain of the Mouse Coronavirus Nucleocapsid Protein. J. Virol. 79: 13285-13297 [Abstract] [Full Text]  
  • Calvo, E., Escors, D., Lopez, J. A., Gonzalez, J. M., Alvarez, A., Arza, E., Enjuanes, L. (2005). Phosphorylation and subcellular localization of transmissible gastroenteritis virus nucleocapsid protein in infected cells. J. Gen. Virol. 86: 2255-2267 [Abstract] [Full Text]  
  • Yu, I-M., Gustafson, C. L. T., Diao, J., Burgner, J. W. II, Li, Z., Zhang, J., Chen, J. (2005). Recombinant Severe Acute Respiratory Syndrome (SARS) Coronavirus Nucleocapsid Protein Forms a Dimer through Its C-terminal Domain. J. Biol. Chem. 280: 23280-23286 [Abstract] [Full Text]  
  • Schelle, B., Karl, N., Ludewig, B., Siddell, S. G., Thiel, V. (2005). Selective Replication of Coronavirus Genomes That Express Nucleocapsid Protein. J. Virol. 79: 6620-6630 [Abstract] [Full Text]  
  • Brunner, J. E., Nguyen, J. H. C., Roehl, H. H., Ho, T. V., Swiderek, K. M., Semler, B. L. (2005). Functional Interaction of Heterogeneous Nuclear Ribonucleoprotein C with Poliovirus RNA Synthesis Initiation Complexes. J. Virol. 79: 3254-3266 [Abstract] [Full Text]  
  • Ito, N., Mossel, E. C., Narayanan, K., Popov, V. L., Huang, C., Inoue, T., Peters, C. J., Makino, S. (2005). Severe Acute Respiratory Syndrome Coronavirus 3a Protein Is a Viral Structural Protein. J. Virol. 79: 3182-3186 [Abstract] [Full Text]  
  • Chen, H., Gill, A., Dove, B. K., Emmett, S. R., Kemp, C. F., Ritchie, M. A., Dee, M., Hiscox, J. A. (2005). Mass Spectroscopic Characterization of the Coronavirus Infectious Bronchitis Virus Nucleoprotein and Elucidation of the Role of Phosphorylation in RNA Binding by Using Surface Plasmon Resonance. J. Virol. 79: 1164-1179 [Abstract] [Full Text]  
  • Huang, Y., Yang, Z.-y., Kong, W.-p., Nabel, G. J. (2004). Generation of Synthetic Severe Acute Respiratory Syndrome Coronavirus Pseudoparticles: Implications for Assembly and Vaccine Production. J. Virol. 78: 12557-12565 [Abstract] [Full Text]  
  • Bosch, B. J., de Haan, C. A. M., Rottier, P. J. M. (2004). Coronavirus Spike Glycoprotein, Extended at the Carboxy Terminus with Green Fluorescent Protein, Is Assembly Competent. J. Virol. 78: 7369-7378 [Abstract] [Full Text]  
  • Buchholz, U. J., Bukreyev, A., Yang, L., Lamirande, E. W., Murphy, B. R., Subbarao, K., Collins, P. L. (2004). Contributions of the structural proteins of severe acute respiratory syndrome coronavirus to protective immunity. Proc. Natl. Acad. Sci. USA 101: 9804-9809 [Abstract] [Full Text]  
  • Chen, Z., Pei, D., Jiang, L., Song, Y., Wang, J., Wang, H., Zhou, D., Zhai, J., Du, Z., Li, B., Qiu, M., Han, Y., Guo, Z., Yang, R. (2004). Antigenicity Analysis of Different Regions of the Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein. Clin. Chem. 50: 988-995 [Abstract] [Full Text]  
  • Lobigs, M., Lee, E. (2004). Inefficient Signalase Cleavage Promotes Efficient Nucleocapsid Incorporation into Budding Flavivirus Membranes. J. Virol. 78: 178-186 [Abstract] [Full Text]  
  • Escors, D., Izeta, A., Capiscol, C., Enjuanes, L. (2003). Transmissible Gastroenteritis Coronavirus Packaging Signal Is Located at the 5' End of the Virus Genome. J. Virol. 77: 7890-7902 [Abstract] [Full Text]  
  • Kuo, L., Masters, P. S. (2003). The Small Envelope Protein E Is Not Essential for Murine Coronavirus Replication. J. Virol. 77: 4597-4608 [Abstract] [Full Text]  
  • Narayanan, K., Chen, C.-J., Maeda, J., Makino, S. (2003). Nucleocapsid-Independent Specific Viral RNA Packaging via Viral Envelope Protein and Viral RNA Signal. J. Virol. 77: 2922-2927 [Abstract] [Full Text]  
  • Kuo, L., Masters, P. S. (2002). Genetic Evidence for a Structural Interaction between the Carboxy Termini of the Membrane and Nucleocapsid Proteins of Mouse Hepatitis Virus. J. Virol. 76: 4987-4999 [Abstract] [Full Text]  
  • Curtis, K. M., Yount, B., Baric, R. S. (2002). Heterologous Gene Expression from Transmissible Gastroenteritis Virus Replicon Particles. J. Virol. 76: 1422-1434 [Abstract] [Full Text]  
  • Escors, D., Camafeita, E., Ortego, J., Laude, H., Enjuanes, L. (2001). Organization of Two Transmissible Gastroenteritis Coronavirus Membrane Protein Topologies within the Virion and Core. J. Virol. 75: 12228-12240 [Abstract] [Full Text]  
  • Verheije, M. H., Kroese, M. V., Rottier, P. J. M., Meulenberg, J. J. M. (2001). Viable porcine arteriviruses with deletions proximal to the 3' end of the genome. J. Gen. Virol. 82: 2607-2614 [Abstract] [Full Text]  
  • Narayanan, K., Makino, S. (2001). Cooperation of an RNA Packaging Signal and a Viral Envelope Protein in Coronavirus RNA Packaging. J. Virol. 75: 9059-9067 [Abstract] [Full Text]  
  • Escors, D., Ortego, J., Laude, H., Enjuanes, L. (2001). The Membrane M Protein Carboxy Terminus Binds to Transmissible Gastroenteritis Coronavirus Core and Contributes to Core Stability. J. Virol. 75: 1312-1324 [Abstract] [Full Text]  
  • Lim, K. P., Liu, D. X. (2001). The Missing Link in Coronavirus Assembly. RETENTION OF THE AVIAN CORONAVIRUS INFECTIOUS BRONCHITIS VIRUS ENVELOPE PROTEIN IN THE PRE-GOLGI COMPARTMENTS AND PHYSICAL INTERACTION BETWEEN THE ENVELOPE AND MEMBRANE PROTEINS. J. Biol. Chem. 276: 17515-17523 [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»