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 Frolova, E.
Right arrow Articles by Frolov, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Frolova, E.
Right arrow Articles by Frolov, I.

 Previous Article  |  Next Article 

Journal of Virology, April 2006, p. 4122-4134, Vol. 80, No. 8
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.8.4122-4134.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Formation of nsP3-Specific Protein Complexes during Sindbis Virus Replication

Elena Frolova,1,2 Rodion Gorchakov,2 Natalia Garmashova,2 Svetlana Atasheva,2 Leoncio A. Vergara,3 and Ilya Frolov2*

Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, Texas 77555-1072,1 Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas 77555-1019,2 Optical Imaging Laboratory, University of Texas Medical Branch, Galveston, Texas 77555-06413

Received 30 November 2005/ Accepted 21 January 2006

Alphaviruses are arthropod-borne viruses (arboviruses) that include a number of important human and animal pathogens. Their replication proceeds in the cytoplasm of infected cells and does not directly depend on nuclei. Alphaviruses encode only four nonstructural proteins that are required for the replication of viral genome and transcription of the subgenomic RNA. However, the replicative enzyme complexes (RCs) appear to include cellular proteins and assemble on cellular organelles. We have developed a set of recombinant Sindbis (SIN) viruses with green fluorescent protein (GFP) insertions in one of the nonstructural proteins, nsP3, to further understand the RCs' genesis and structure. We studied the assembly of nsP3/GFP-containing protein complexes at different stages of infection and isolated a combination of cellular proteins that are associated with SIN nsP3. We demonstrated the following. (i) SIN nsP3 can tolerate the insertion of GFP into different fragments of the coding sequence; the designed recombinant viruses are viable, and their replication leads to the assembly of nsP3/GFP chimeric proteins into gradually developing, higher-order structures differently organized at early and late times postinfection. (ii) At late times postinfection, nsP3 is assembled into complexes of similar sizes, which appear to be bound to cytoskeleton filaments and can aggregate into larger structures. (iii) Protein complexes that are associated with nsP3/GFP contain a high concentration of cytoskeleton proteins, chaperones, elongation factor 1A, heterogeneous nuclear ribonucleoproteins, 14-3-3 proteins, and some of the ribosomal proteins. These proteins are proposed to be essential for SIN RC formation and/or functioning.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX 77555-1019. Phone: (409) 772-2327. Fax: (409) 772-5065. E-mail: ivfrolov{at}UTMB.edu.

Journal of Virology, April 2006, p. 4122-4134, Vol. 80, No. 8
0022-538X/06/$08.00+0     doi:10.1128/JVI.80.8.4122-4134.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Kulasegaran-Shylini, R., Atasheva, S., Gorenstein, D. G., Frolov, I. (2009). Structural and Functional Elements of the Promoter Encoded by the 5' Untranslated Region of the Venezuelan Equine Encephalitis Virus Genome. J. Virol. 83: 8327-8339 [Abstract] [Full Text]  
  • Lin, J.-Y., Shih, S.-R., Pan, M., Li, C., Lue, C.-F., Stollar, V., Li, M.-L. (2009). hnRNP A1 Interacts with the 5' Untranslated Regions of Enterovirus 71 and Sindbis Virus RNA and Is Required for Viral Replication. J. Virol. 83: 6106-6114 [Abstract] [Full Text]  
  • Gorchakov, R., Garmashova, N., Frolova, E., Frolov, I. (2008). Different Types of nsP3-Containing Protein Complexes in Sindbis Virus-Infected Cells. J. Virol. 82: 10088-10101 [Abstract] [Full Text]  
  • Gorchakov, R., Frolova, E., Sawicki, S., Atasheva, S., Sawicki, D., Frolov, I. (2008). A New Role for ns Polyprotein Cleavage in Sindbis Virus Replication. J. Virol. 82: 6218-6231 [Abstract] [Full Text]  
  • Weeks, S. A., Miller, D. J. (2008). The Heat Shock Protein 70 Cochaperone YDJ1 Is Required for Efficient Membrane-Specific Flock House Virus RNA Replication Complex Assembly and Function in Saccharomyces cerevisiae. J. Virol. 82: 2004-2012 [Abstract] [Full Text]  
  • Atasheva, S., Gorchakov, R., English, R., Frolov, I., Frolova, E. (2007). Development of Sindbis Viruses Encoding nsP2/GFP Chimeric Proteins and Their Application for Studying nsP2 Functioning. J. Virol. 81: 5046-5057 [Abstract] [Full Text]  
  • Tamberg, N., Lulla, V., Fragkoudis, R., Lulla, A., Fazakerley, J. K., Merits, A. (2007). Insertion of EGFP into the replicase gene of Semliki Forest virus results in a novel, genetically stable marker virus. J. Gen. Virol. 88: 1225-1230 [Abstract] [Full Text]  
  • Cristea, I. M., Carroll, J.-W. N., Rout, M. P., Rice, C. M., Chait, B. T., MacDonald, M. R. (2006). Tracking and Elucidating Alphavirus-Host Protein Interactions. J. Biol. Chem. 281: 30269-30278 [Abstract] [Full Text]  
  • Garmashova, N., Gorchakov, R., Frolova, E., Frolov, I. (2006). Sindbis Virus Nonstructural Protein nsP2 Is Cytotoxic and Inhibits Cellular Transcription.. J. Virol. 80: 5686-5696 [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»