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 Bost, A. G.
Right arrow Articles by Denison, M. R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Bost, A. G.
Right arrow Articles by Denison, M. R.

 Previous Article  |  Next Article 

Journal of Virology, April 2000, p. 3379-3387, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Four Proteins Processed from the Replicase Gene Polyprotein of Mouse Hepatitis Virus Colocalize in the Cell Periphery and Adjacent to Sites of Virion Assembly

Anne Gibson Bost,1 Robert H. Carnahan,2 Xiao Tao Lu,3 and Mark R. Denison1,3,*

Department of Microbiology and Immunology,1 Department of Cell Biology,2 and Department of Pediatrics and the Elizabeth B. Lamb Center for Pediatric Research,3 Vanderbilt University, Nashville, Tennessee 37232

Received 28 September 1999/Accepted 21 December 1999

The replicase gene (gene 1) of the coronavirus mouse hepatitis virus (MHV) encodes two co-amino-terminal polyproteins presumed to incorporate all the virus-encoded proteins necessary for viral RNA synthesis. The polyproteins are cotranslationally processed by viral proteinases into at least 15 mature proteins, including four predicted cleavage products of less than 25 kDa that together would comprise the final 59 kDa of protein translated from open reading frame 1a. Monospecific antibodies directed against the four distinct domains detected proteins of 10, 12, and 15 kDa (p1a-10, p1a-12, and p1a-15) in MHV-A59-infected DBT cells, in addition to a previously identified 22-kDa protein (p1a-22). When infected cells were probed by immunofluorescence laser confocal microscopy, p1a-10, -22, -12, and -15 were detected in discrete foci that were prominent in the perinuclear region but were widely distributed throughout the cytoplasm as well. Dual-labeling experiments demonstrated colocalization of the majority of p1a-22 in replication complexes with the helicase, nucleocapsid, and 3C-like proteinase, as well as with p1a-10, -12, and -15. p1a-22 was also detected in separate foci adjacent to the replication complexes. The majority of complexes containing the gene 1 proteins were distinct from sites of accumulation of the M assembly protein. However, in perinuclear regions the gene 1 proteins and nucleocapsid were intercalated with sites of M protein localization. These results demonstrate that the complexes known to be involved in RNA synthesis contain multiple gene 1 proteins and are closely associated with structural proteins at presumed sites of virion assembly.

* Corresponding author. Mailing address: Department of Pediatrics, Vanderbilt University Medical Center, D7235 MCN, Nashville, TN 37232-2581. Phone: (615) 343-9881. Fax: (615) 343-9723. E-mail: mark.denison{at}mcmail.vanderbilt.edu.

Journal of Virology, April 2000, p. 3379-3387, Vol. 74, No. 7
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Baliji, S., Cammer, S. A., Sobral, B., Baker, S. C. (2009). Detection of Nonstructural Protein 6 in Murine Coronavirus-Infected Cells and Analysis of the Transmembrane Topology by Using Bioinformatics and Molecular Approaches. J. Virol. 83: 6957-6962 [Abstract] [Full Text]  
  • Miknis, Z. J., Donaldson, E. F., Umland, T. C., Rimmer, R. A., Baric, R. S., Schultz, L. W. (2009). Severe Acute Respiratory Syndrome Coronavirus nsp9 Dimerization Is Essential for Efficient Viral Growth. J. Virol. 83: 3007-3018 [Abstract] [Full Text]  
  • Oostra, M., Hagemeijer, M. C., van Gent, M., Bekker, C. P. J., te Lintelo, E. G., Rottier, P. J. M., de Haan, C. A. M. (2008). Topology and Membrane Anchoring of the Coronavirus Replication Complex: Not All Hydrophobic Domains of nsp3 and nsp6 Are Membrane Spanning. J. Virol. 82: 12392-12405 [Abstract] [Full Text]  
  • van Hemert, M. J., de Wilde, A. H., Gorbalenya, A. E., Snijder, E. J. (2008). The in Vitro RNA Synthesizing Activity of the Isolated Arterivirus Replication/Transcription Complex Is Dependent on a Host Factor. J. Biol. Chem. 283: 16525-16536 [Abstract] [Full Text]  
  • Zust, R., Miller, T. B., Goebel, S. J., Thiel, V., Masters, P. S. (2008). Genetic Interactions between an Essential 3' cis-Acting RNA Pseudoknot, Replicase Gene Products, and the Extreme 3' End of the Mouse Coronavirus Genome. J. Virol. 82: 1214-1228 [Abstract] [Full Text]  
  • Sparks, J. S., Lu, X., Denison, M. R. (2007). Genetic Analysis of Murine Hepatitis Virus nsp4 in Virus Replication. J. Virol. 81: 12554-12563 [Abstract] [Full Text]  
  • Oostra, M., te Lintelo, E. G., Deijs, M., Verheije, M. H., Rottier, P. J. M., de Haan, C. A. M. (2007). Localization and Membrane Topology of Coronavirus Nonstructural Protein 4: Involvement of the Early Secretory Pathway in Replication. J. Virol. 81: 12323-12336 [Abstract] [Full Text]  
  • Deming, D. J., Graham, R. L., Denison, M. R., Baric, R. S. (2007). Processing of Open Reading Frame 1a Replicase Proteins nsp7 to nsp10 in Murine Hepatitis Virus Strain A59 Replication. J. Virol. 81: 10280-10291 [Abstract] [Full Text]  
  • Donaldson, E. F., Graham, R. L., Sims, A. C., Denison, M. R., Baric, R. S. (2007). Analysis of Murine Hepatitis Virus Strain A59 Temperature-Sensitive Mutant TS-LA6 Suggests that nsp10 Plays a Critical Role in Polyprotein Processing. J. Virol. 81: 7086-7098 [Abstract] [Full Text]  
  • Donaldson, E. F., Sims, A. C., Graham, R. L., Denison, M. R., Baric, R. S. (2007). Murine Hepatitis Virus Replicase Protein nsp10 Is a Critical Regulator of Viral RNA Synthesis. J. Virol. 81: 6356-6368 [Abstract] [Full Text]  
  • Cai, Y., Liu, Y., Zhang, X. (2007). Suppression of Coronavirus Replication by Inhibition of the MEK Signaling Pathway. J. Virol. 81: 446-456 [Abstract] [Full Text]  
  • Sawicki, S. G., Sawicki, D. L., Siddell, S. G. (2007). A Contemporary View of Coronavirus Transcription. J. Virol. 81: 20-29 [Full Text]  
  • van Aken, D., Zevenhoven-Dobbe, J., Gorbalenya, A. E., Snijder, E. J. (2006). Proteolytic maturation of replicase polyprotein pp1a by the nsp4 main proteinase is essential for equine arteritis virus replication and includes internal cleavage of nsp7. J. Gen. Virol. 87: 3473-3482 [Abstract] [Full Text]  
  • Graham, R. L., Denison, M. R. (2006). Replication of Murine Hepatitis Virus Is Regulated by Papain-Like Proteinase 1 Processing of Nonstructural Proteins 1, 2, and 3. J. Virol. 80: 11610-11620 [Abstract] [Full Text]  
  • Joseph, J. S., Saikatendu, K. S., Subramanian, V., Neuman, B. W., Brooun, A., Griffith, M., Moy, K., Yadav, M. K., Velasquez, J., Buchmeier, M. J., Stevens, R. C., Kuhn, P. (2006). Crystal structure of nonstructural protein 10 from the severe acute respiratory syndrome coronavirus reveals a novel fold with two zinc-binding motifs.. J. Virol. 80: 7894-7901 [Abstract] [Full Text]  
  • Su, D., Lou, Z., Sun, F., Zhai, Y., Yang, H., Zhang, R., Joachimiak, A., Zhang, X. C., Bartlam, M., Rao, Z. (2006). Dodecamer Structure of Severe Acute Respiratory Syndrome Coronavirus Nonstructural Protein nsp10.. J. Virol. 80: 7902-7908 [Abstract] [Full Text]  
  • Graham, R. L., Sims, A. C., Brockway, S. M., Baric, R. S., Denison, M. R. (2005). The nsp2 Replicase Proteins of Murine Hepatitis Virus and Severe Acute Respiratory Syndrome Coronavirus Are Dispensable for Viral Replication. J. Virol. 79: 13399-13411 [Abstract] [Full Text]  
  • Peti, W., Johnson, M. A., Herrmann, T., Neuman, B. W., Buchmeier, M. J., Nelson, M., Joseph, J., Page, R., Stevens, R. C., Kuhn, P., Wuthrich, K. (2005). Structural Genomics of the Severe Acute Respiratory Syndrome Coronavirus: Nuclear Magnetic Resonance Structure of the Protein nsP7. J. Virol. 79: 12905-12913 [Abstract] [Full Text]  
  • Brockway, S. M., Lu, X. T., Peters, T. R., Dermody, T. S., Denison, M. R. (2004). Intracellular Localization and Protein Interactions of the Gene 1 Protein p28 during Mouse Hepatitis Virus Replication. J. Virol. 78: 11551-11562 [Abstract] [Full Text]  
  • Prentice, E., McAuliffe, J., Lu, X., Subbarao, K., Denison, M. R. (2004). Identification and Characterization of Severe Acute Respiratory Syndrome Coronavirus Replicase Proteins. J. Virol. 78: 9977-9986 [Abstract] [Full Text]  
  • Tan, Y.-J., Teng, E., Shen, S., Tan, T. H. P., Goh, P.-Y., Fielding, B. C., Ooi, E.-E., Tan, H.-C., Lim, S. G., Hong, W. (2004). A Novel Severe Acute Respiratory Syndrome Coronavirus Protein, U274, Is Transported to the Cell Surface and Undergoes Endocytosis. J. Virol. 78: 6723-6734 [Abstract] [Full Text]  
  • Ivanov, K. A., Thiel, V., Dobbe, J. C., van der Meer, Y., Snijder, E. J., Ziebuhr, J. (2004). Multiple Enzymatic Activities Associated with Severe Acute Respiratory Syndrome Coronavirus Helicase. J. Virol. 78: 5619-5632 [Abstract] [Full Text]  
  • Egloff, M.-P., Ferron, F., Campanacci, V., Longhi, S., Rancurel, C., Dutartre, H., Snijder, E. J., Gorbalenya, A. E., Cambillau, C., Canard, B. (2004). The severe acute respiratory syndrome-coronavirus replicative protein nsp9 is a single-stranded RNA-binding subunit unique in the RNA virus world. Proc. Natl. Acad. Sci. USA 101: 3792-3796 [Abstract] [Full Text]  
  • Brockway, S. M., Clay, C. T., Lu, X. T., Denison, M. R. (2003). Characterization of the Expression, Intracellular Localization, and Replication Complex Association of the Putative Mouse Hepatitis Virus RNA-Dependent RNA Polymerase. J. Virol. 77: 10515-10527 [Abstract] [Full Text]  
  • Thiel, V., Ivanov, K. A., Putics, A., Hertzig, T., Schelle, B., Bayer, S., Weissbrich, B., Snijder, E. J., Rabenau, H., Doerr, H. W., Gorbalenya, A. E., Ziebuhr, J. (2003). Mechanisms and enzymes involved in SARS coronavirus genome expression. J. Gen. Virol. 84: 2305-2315 [Abstract] [Full Text]  
  • Yount, B., Denison, M. R., Weiss, S. R., Baric, R. S. (2002). Systematic Assembly of a Full-Length Infectious cDNA of Mouse Hepatitis Virus Strain A59. J. Virol. 76: 11065-11078 [Abstract] [Full Text]  
  • Ng, L. F. P., Liu, D. X. (2002). Membrane Association and Dimerization of a Cysteine-Rich, 16-Kilodalton Polypeptide Released from the C-Terminal Region of the Coronavirus Infectious Bronchitis Virus 1a Polyprotein. J. Virol. 76: 6257-6267 [Abstract] [Full Text]  
  • Gosert, R., Kanjanahaluethai, A., Egger, D., Bienz, K., Baker, S. C. (2002). RNA Replication of Mouse Hepatitis Virus Takes Place at Double-Membrane Vesicles. J. Virol. 76: 3697-3708 [Abstract] [Full Text]  
  • Kanjanahaluethai, A., Baker, S. C. (2000). Identification of Mouse Hepatitis Virus Papain-Like Proteinase 2 Activity. J. Virol. 74: 7911-7921 [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»