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 Tellinghuisen, T. L.
Right arrow Articles by Kuhn, R. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Tellinghuisen, T. L.
Right arrow Articles by Kuhn, R. J.

 Previous Article  |  Next Article 

Journal of Virology, March 2001, p. 2810-2817, Vol. 75, No. 6
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.6.2810-2817.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

In Vitro Assembly of Sindbis Virus Core-Like Particles from Cross-Linked Dimers of Truncated and Mutant Capsid Proteins

Timothy L. Tellinghuisen, Rushika Perera, and Richard J. Kuhn*

Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907

Received 6 September 2000/Accepted 19 December 2000

A nucleic acid-bound capsid protein dimer was previously identified using a Sindbis virus in vitro nucleocapsid assembly system and cross-linking reagents. Cross-link mapping, in combination with a model of the nucleocapsid core, suggested that this dimer contained one monomer from each of two adjacent capsomeres. This intercapsomere dimer is believed to be the initial intermediate in the nucleocapsid core assembly mechanism. This paper presents the purification of cross-linked dimers of a truncated capsid protein and the partial purification of cross-linked dimers of a full-length assembly-defective mutant. The assembly of core-like particles from these cross-linked capsid protein dimers is demonstrated. Core-like particles generated from cross-linked full-length mutant CP(19-264)L52D were examined by electron microscopy and appeared to have a morphology similar to that of wild-type in vitro-assembled core-like particles, although a slight size difference was often visible. Truncated cross-linked CP(81-264) dimers generated core-like particles as well. These core-like particles could subsequently be disassembled when reversible cross-linking reagents were used to form the dimers. The ability of the covalent intercapsomere cross-link to rescue capsid proteins with assembly defects or truncations in the amino-terminal region of the capsid protein supports the previous model of assembly and suggests a possible role for the amino-terminal region of the protein.

* Corresponding author. Mailing address: Department of Biological Sciences, Lilly Hall of Life Sciences, Purdue University, West Lafayette, IN 47907-1392. Phone: (765) 494-1164. Fax: (765) 496-1189. E-mail: rjkuhn{at}bragg.bio.purdue.edu.

Journal of Virology, March 2001, p. 2810-2817, Vol. 75, No. 6
0022-538X/01/$04.00+0   DOI: 10.1128/JVI.75.6.2810-2817.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Warrier, R., Linger, B. R., Golden, B. L., Kuhn, R. J. (2008). Role of Sindbis Virus Capsid Protein Region II in Nucleocapsid Core Assembly and Encapsidation of Genomic RNA. J. Virol. 82: 4461-4470 [Abstract] [Full Text]  
  • Bu, W., Carroll, K. D., Palmeri, D., Lukac, D. M. (2007). Kaposi's Sarcoma-Associated Herpesvirus/Human Herpesvirus 8 ORF50/Rta Lytic Switch Protein Functions as a Tetramer. J. Virol. 81: 5788-5806 [Abstract] [Full Text]  
  • Aguilar, P. V., Weaver, S. C., Basler, C. F. (2007). Capsid Protein of Eastern Equine Encephalitis Virus Inhibits Host Cell Gene Expression. J. Virol. 81: 3866-3876 [Abstract] [Full Text]  
  • Hong, E. M., Perera, R., Kuhn, R. J. (2006). Alphavirus capsid protein helix I controls a checkpoint in nucleocapsid core assembly.. J. Virol. 80: 8848-8855 [Abstract] [Full Text]  
  • Boulant, S., Vanbelle, C., Ebel, C., Penin, F., Lavergne, J.-P. (2005). Hepatitis C Virus Core Protein Is a Dimeric Alpha-Helical Protein Exhibiting Membrane Protein Features. J. Virol. 79: 11353-11365 [Abstract] [Full Text]  
  • Ma, Y. M., Vogt, V. M. (2004). Nucleic Acid Binding-Induced Gag Dimerization in the Assembly of Rous Sarcoma Virus Particles In Vitro. J. Virol. 78: 52-60 [Abstract] [Full Text]  
  • LINGER, B. R., KUNOVSKA, L., KUHN, R. J., GOLDEN, B. L. (2004). Sindbis virus nucleocapsid assembly: RNA folding promotes capsid protein dimerization. RNA 10: 128-138 [Abstract] [Full Text]  
  • Hernandez, R., Sinodis, C., Horton, M., Ferreira, D., Yang, C., Brown, D. T. (2003). Deletions in the Transmembrane Domain of a Sindbis Virus Glycoprotein Alter Virus Infectivity, Stability, and Host Range. J. Virol. 77: 12710-12719 [Abstract] [Full Text]  
  • Perera, R., Navaratnarajah, C., Kuhn, R. J. (2003). A Heterologous Coiled Coil Can Substitute for Helix I of the Sindbis Virus Capsid Protein. J. Virol. 77: 8345-8353 [Abstract] [Full Text]  
  • Zhang, W., Mukhopadhyay, S., Pletnev, S. V., Baker, T. S., Kuhn, R. J., Rossmann, M. G. (2002). Placement of the Structural Proteins in Sindbis Virus. J. Virol. 76: 11645-11658 [Abstract] [Full Text]  
  • Ma, Y. M., Vogt, V. M. (2002). Rous Sarcoma Virus Gag Protein-Oligonucleotide Interaction Suggests a Critical Role for Protein Dimer Formation in Assembly. J. Virol. 76: 5452-5462 [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»