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 Ng, L. F. P.
Right arrow Articles by Liu, D. X.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ng, L. F. P.
Right arrow Articles by Liu, D. X.

 Previous Article  |  Next Article 

Journal of Virology, June 2002, p. 6257-6267, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6257-6267.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

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

Lisa F. P. Ng1 and D. X. Liu1,2*

Institute of Molecular Agrobiology, The National University of Singapore, Singapore 117604,1 School of Biological Sciences, Nanyang Technological University, Singapore 6376162

Received 6 December 2001/ Accepted 20 March 2002

More than 10 mature proteins processed from coronavirus gene 1-encoded polyproteins have been identified in virus-infected cells. Here, we report the identification of the most C-terminal cleavage product of the 1a polyprotein as a 16-kDa protein in infectious bronchitis virus-infected Vero cells. Indirect immunofluorescence demonstrated that the protein exhibits a distinct perinuclear punctate staining pattern, suggesting that it is associated with cellular membranes. Positive staining observed on nonpermeabilized cells indicates that the protein may get transported to the cell surface, but no secretion of the protein out of the cells was observed. Treatment of the membrane fraction prepared from cells expressing the 16-kDa protein with Triton X-100, a high pH, and a high concentration of salts showed that the protein may be tightly associated with intracellular membranes. Dual-labeling experiments demonstrated that the 16-kDa protein colocalized with the 5'-bromouridine 5'-triphosphate-labeled viral RNA, suggesting that it may be associated with the viral replication machinery. Sequence comparison of the 16-kDa protein with the equivalent products of other coronaviruses showed multiple conserved cysteine residues, and site-directed mutagenesis studies revealed that these conserved residues may contribute to dimerization of the 16-kDa protein. Furthermore, increased accumulation of the 16-kDa protein upon stimulation with epidermal growth factor was observed, providing preliminary evidence that the protein might be involved in the growth factor signaling pathway.

* Corresponding author. Mailing address: School of Biological Sciences, Nanyang Technological University, 1 Nanyang Walk, Block 5, Level 3, Singapore 637616. Phone: 65-872-7000. Fax: 65-872-7007. E-mail: dxliu{at}ntu.edu.sg.

Journal of Virology, June 2002, p. 6257-6267, Vol. 76, No. 12
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.12.6257-6267.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Wang, X., Liao, Y., Yap, P. L., Png, K. J., Tam, J. P., Liu, D. X. (2009). Inhibition of Protein Kinase R Activation and Upregulation of GADD34 Expression Play a Synergistic Role in Facilitating Coronavirus Replication by Maintaining De Novo Protein Synthesis in Virus-Infected Cells. J. Virol. 83: 12462-12472 [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]  
  • 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]  
  • Pendleton, A. R., Machamer, C. E. (2005). Infectious Bronchitis Virus 3a Protein Localizes to a Novel Domain of the Smooth Endoplasmic Reticulum. J. Virol. 79: 6142-6151 [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]  
  • Ng, L. F. P., Wong, M., Koh, S., Ooi, E.-E., Tang, K.-F., Leong, H.-N., Ling, A.-E., Agathe, L. V., Tan, J., Liu, E. T., Ren, E.-C., Ng, L.-C., Hibberd, M. L. (2004). Detection of Severe Acute Respiratory Syndrome Coronavirus in Blood of Infected Patients. J. Clin. Microbiol. 42: 347-350 [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»