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
Right arrow Citation Map
Services
Right arrow E-mail this article to a friend
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 Dimitrova, M.
Right arrow Articles by Schuster, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Dimitrova, M.
Right arrow Articles by Schuster, C.

 Previous Article  |  Next Article 

Journal of Virology, May 2003, p. 5401-5414, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5401-5414.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Protein-Protein Interactions between Hepatitis C Virus Nonstructural Proteins

Maria Dimitrova, Isabelle Imbert, Marie Paule Kieny, and Catherine Schuster*

INSERM UMR_U544, Institut de Virologie, 67000 Strasbourg, France

Received 19 September 2002/ Accepted 4 February 2003

Replication of the hepatitis C virus (HCV) genome has been proposed to take place close to the membrane of the endoplasmic reticulum in membrane-associated replicase complexes, as is the case with several other plus-strand RNA viruses, such as poliovirus and flaviviruses. The most obvious benefits of this property are the possibility of coupling functions residing in different polypeptidic chains and the sequestration of viral proteins and nucleic acids in a distinct cytoplasmic compartment with high local concentrations of viral components. Indeed, HCV nonstructural (NS) proteins were clearly colocalized in association with membranes derived from the endoplasmic reticulum. This observation, together with the demonstration of the existence of several physical interactions between HCV NS proteins, supports the idea of assembly of a highly ordered multisubunit protein complex(es) probably involved in the replication of the viral genome. The objective of this study, therefore, was to examine all potential interactions between HCV NS proteins which could result in the formation of a replication complex(es). We identified several interacting viral partners by using a glutathione S-transferase pull-down assay, by in vitro and ex vivo coimmunoprecipitation experiments in adenovirus-infected Huh-7 cells allowing the expression of HCV NS proteins, and, finally, by using the yeast two-hybrid system. In addition, by confocal laser scanning microscopy, NS proteins were clearly shown to colocalize when expressed together in Huh-7 cells. We have been able to demonstrate the existence of a complex network of interactions implicating all six NS proteins. Our observations confirm previously described associations and identify several novel homo- and heterodimerizations.

* Corresponding author. Mailing address: INSERM UMR_U544, Institut de Virologie, 3, rue Koeberlé, 67000 Strasbourg, France. Phone: 33 3 90 24 37 41. Fax: 33 3 90 24 37 23. E-mail: catherine.schuster{at}viro-ulp.u-strasbg.fr.

Journal of Virology, May 2003, p. 5401-5414, Vol. 77, No. 9
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.9.5401-5414.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Bungyoku, Y., Shoji, I., Makine, T., Adachi, T., Hayashida, K., Nagano-Fujii, M., Ide, Y.-H., Deng, L., Hotta, H. (2009). Efficient production of infectious hepatitis C virus with adaptive mutations in cultured hepatoma cells. J. Gen. Virol. 90: 1681-1691 [Abstract] [Full Text]  
  • Liu, Z., Yang, F., Robotham, J. M., Tang, H. (2009). Critical Role of Cyclophilin A and Its Prolyl-Peptidyl Isomerase Activity in the Structure and Function of the Hepatitis C Virus Replication Complex. J. Virol. 83: 6554-6565 [Abstract] [Full Text]  
  • Love, R. A., Brodsky, O., Hickey, M. J., Wells, P. A., Cronin, C. N. (2009). Crystal Structure of a Novel Dimeric Form of NS5A Domain I Protein from Hepatitis C Virus. J. Virol. 83: 4395-4403 [Abstract] [Full Text]  
  • Schregel, V., Jacobi, S., Penin, F., Tautz, N. (2009). Hepatitis C virus NS2 is a protease stimulated by cofactor domains in NS3. Proc. Natl. Acad. Sci. USA 106: 5342-5347 [Abstract] [Full Text]  
  • Jones, D. M., Patel, A. H., Targett-Adams, P., McLauchlan, J. (2009). The Hepatitis C Virus NS4B Protein Can trans-Complement Viral RNA Replication and Modulates Production of Infectious Virus. J. Virol. 83: 2163-2177 [Abstract] [Full Text]  
  • Jennings, T. A., Mackintosh, S. G., Harrison, M. K., Sikora, D., Sikora, B., Dave, B., Tackett, A. J., Cameron, C. E., Raney, K. D. (2009). NS3 Helicase from the Hepatitis C Virus Can Function as a Monomer or Oligomer Depending on Enzyme and Substrate Concentrations. J. Biol. Chem. 284: 4806-4814 [Abstract] [Full Text]  
  • Serebrov, V., Beran, R. K. F., Pyle, A. M. (2009). Establishing a Mechanistic Basis for the Large Kinetic Steps of the NS3 Helicase. J. Biol. Chem. 284: 2512-2521 [Abstract] [Full Text]  
  • Paredes, A. M., Blight, K. J. (2008). A Genetic Interaction between Hepatitis C Virus NS4B and NS3 Is Important for RNA Replication. J. Virol. 82: 10671-10683 [Abstract] [Full Text]  
  • Ma, Y., Yates, J., Liang, Y., Lemon, S. M., Yi, M. (2008). NS3 Helicase Domains Involved in Infectious Intracellular Hepatitis C Virus Particle Assembly. J. Virol. 82: 7624-7639 [Abstract] [Full Text]  
  • Wolf, M., Dimitrova, M., Baumert, T. F., Schuster, C. (2008). The major form of hepatitis C virus alternate reading frame protein is suppressed by core protein expression. Nucleic Acids Res 36: 3054-3064 [Abstract] [Full Text]  
  • Sikora, B., Chen, Y., Lichti, C. F., Harrison, M. K., Jennings, T. A., Tang, Y., Tackett, A. J., Jordan, J. B., Sakon, J., Cameron, C. E., Raney, K. D. (2008). Hepatitis C Virus NS3 Helicase Forms Oligomeric Structures That Exhibit Optimal DNA Unwinding Activity in Vitro. J. Biol. Chem. 283: 11516-11525 [Abstract] [Full Text]  
  • Tang, W., Lazaro, C. A., Campbell, J. S., Parks, W. T., Katze, M. G., Fausto, N. (2007). Responses of Nontransformed Human Hepatocytes to Conditional Expression of Full-Length Hepatitis C Virus Open Reading Frame. Am. J. Pathol. 171: 1831-1846 [Abstract] [Full Text]  
  • Lindenbach, B. D., Pragai, B. M., Montserret, R., Beran, R. K. F., Pyle, A. M., Penin, F., Rice, C. M. (2007). The C Terminus of Hepatitis C Virus NS4A Encodes an Electrostatic Switch That Regulates NS5A Hyperphosphorylation and Viral Replication. J. Virol. 81: 8905-8918 [Abstract] [Full Text]  
  • Blight, K. J. (2007). Allelic Variation in the Hepatitis C Virus NS4B Protein Dramatically Influences RNA Replication. J. Virol. 81: 5724-5736 [Abstract] [Full Text]  
  • Binder, M., Quinkert, D., Bochkarova, O., Klein, R., Kezmic, N., Bartenschlager, R., Lohmann, V. (2007). Identification of Determinants Involved in Initiation of Hepatitis C Virus RNA Synthesis by Using Intergenotypic Replicase Chimeras. J. Virol. 81: 5270-5283 [Abstract] [Full Text]  
  • Lee, H., Liu, Y., Mejia, E., Paul, A. V., Wimmer, E. (2006). The C-Terminal Hydrophobic Domain of Hepatitis C Virus RNA Polymerase NS5B Can Be Replaced with a Heterologous Domain of Poliovirus Protein 3A. J. Virol. 80: 11343-11354 [Abstract] [Full Text]  
  • Lundin, M., Lindstrom, H., Gronwall, C., Persson, M. A. A. (2006). Dual topology of the processed hepatitis C virus protein NS4B is influenced by the NS5A protein.. J. Gen. Virol. 87: 3263-3272 [Abstract] [Full Text]  
  • Lionnet, T., Dawid, A., Bigot, S., Barre, F.-X., Saleh, O. A., Heslot, F., Allemand, J.-F., Bensimon, D., Croquette, V. (2006). DNA mechanics as a tool to probe helicase and translocase activity. Nucleic Acids Res 34: 4232-4244 [Abstract] [Full Text]  
  • Umareddy, I., Chao, A., Sampath, A., Gu, F., Vasudevan, S. G. (2006). Dengue virus NS4B interacts with NS3 and dissociates it from single-stranded RNA. J. Gen. Virol. 87: 2605-2614 [Abstract] [Full Text]  
  • Yu, G.-Y., Lee, K.-J., Gao, L., Lai, M. M. C. (2006). Palmitoylation and Polymerization of Hepatitis C Virus NS4B Protein. J. Virol. 80: 6013-6023 [Abstract] [Full Text]  
  • Shimakami, T., Honda, M., Kusakawa, T., Murata, T., Shimotohno, K., Kaneko, S., Murakami, S. (2006). Effect of Hepatitis C Virus (HCV) NS5B-Nucleolin Interaction on HCV Replication with HCV Subgenomic Replicon. J. Virol. 80: 3332-3340 [Abstract] [Full Text]  
  • McCormick, C. J., Maucourant, S., Griffin, S., Rowlands, D. J., Harris, M. (2006). Tagging of NS5A expressed from a functional hepatitis C virus replicon.. J. Gen. Virol. 87: 635-640 [Abstract] [Full Text]  
  • Paeshuyse, J., Leyssen, P., Mabery, E., Boddeker, N., Vrancken, R., Froeyen, M., Ansari, I. H., Dutartre, H., Rozenski, J., Gil, L. H. V. G., Letellier, C., Lanford, R., Canard, B., Koenen, F., Kerkhofs, P., Donis, R. O., Herdewijn, P., Watson, J., De Clercq, E., Puerstinger, G., Neyts, J. (2006). A Novel, Highly Selective Inhibitor of Pestivirus Replication That Targets the Viral RNA-Dependent RNA Polymerase. J. Virol. 80: 149-160 [Abstract] [Full Text]  
  • Quinkert, D., Bartenschlager, R., Lohmann, V. (2005). Quantitative Analysis of the Hepatitis C Virus Replication Complex. J. Virol. 79: 13594-13605 [Abstract] [Full Text]  
  • Huang, L., Hwang, J., Sharma, S. D., Hargittai, M. R. S., Chen, Y., Arnold, J. J., Raney, K. D., Cameron, C. E. (2005). Hepatitis C Virus Nonstructural Protein 5A (NS5A) Is an RNA-binding Protein. J. Biol. Chem. 280: 36417-36428 [Abstract] [Full Text]  
  • Gosert, R., Jendrsczok, W., Berke, J. M., Brass, V., Blum, H. E., Moradpour, D. (2005). Characterization of Nonstructural Protein Membrane Anchor Deletion Mutants Expressed in the Context of the Hepatitis C Virus Polyprotein. J. Virol. 79: 7911-7917 [Abstract] [Full Text]  
  • Ferreon, J. C., Ferreon, A. C. M., Li, K., Lemon, S. M. (2005). Molecular Determinants of TRIF Proteolysis Mediated by the Hepatitis C Virus NS3/4A Protease. J. Biol. Chem. 280: 20483-20492 [Abstract] [Full Text]  
  • Benzaghou, I., Bougie, I., Bisaillon, M. (2004). Effect of Metal Ion Binding on the Structural Stability of the Hepatitis C Virus RNA Polymerase. J. Biol. Chem. 279: 49755-49761 [Abstract] [Full Text]  
  • Kim, S.-J., Kim, J.-H., Kim, Y.-G., Lim, H.-S., Oh, J.-W. (2004). Protein Kinase C-related Kinase 2 Regulates Hepatitis C Virus RNA Polymerase Function by Phosphorylation. J. Biol. Chem. 279: 50031-50041 [Abstract] [Full Text]  
  • Lee, H., Shin, H., Wimmer, E., Paul, A. V. (2004). cis-Acting RNA Signals in the NS5B C-Terminal Coding Sequence of the Hepatitis C Virus Genome. J. Virol. 78: 10865-10877 [Abstract] [Full Text]  
  • Elazar, M., Liu, P., Rice, C. M., Glenn, J. S. (2004). An N-Terminal Amphipathic Helix in Hepatitis C Virus (HCV) NS4B Mediates Membrane Association, Correct Localization of Replication Complex Proteins, and HCV RNA Replication. J. Virol. 78: 11393-11400 [Abstract] [Full Text]  
  • Penin, F., Brass, V., Appel, N., Ramboarina, S., Montserret, R., Ficheux, D., Blum, H. E., Bartenschlager, R., Moradpour, D. (2004). Structure and Function of the Membrane Anchor Domain of Hepatitis C Virus Nonstructural Protein 5A. J. Biol. Chem. 279: 40835-40843 [Abstract] [Full Text]  
  • Macdonald, A., Harris, M. (2004). Hepatitis C virus NS5A: tales of a promiscuous protein. J. Gen. Virol. 85: 2485-2502 [Abstract] [Full Text]  
  • Moradpour, D., Evans, M. J., Gosert, R., Yuan, Z., Blum, H. E., Goff, S. P., Lindenbach, B. D., Rice, C. M. (2004). Insertion of Green Fluorescent Protein into Nonstructural Protein 5A Allows Direct Visualization of Functional Hepatitis C Virus Replication Complexes. J. Virol. 78: 7400-7409 [Abstract] [Full Text]  
  • Ma, Y., Shimakami, T., Luo, H., Hayashi, N., Murakami, S. (2004). Mutational Analysis of Hepatitis C Virus NS5B in the Subgenomic Replicon Cell Culture. J. Biol. Chem. 279: 25474-25482 [Abstract] [Full Text]  
  • Choi, K. H., Groarke, J. M., Young, D. C., Kuhn, R. J., Smith, J. L., Pevear, D. C., Rossmann, M. G. (2004). The structure of the RNA-dependent RNA polymerase from bovine viral diarrhea virus establishes the role of GTP in de novo initiation. Proc. Natl. Acad. Sci. USA 101: 4425-4430 [Abstract] [Full Text]  
  • Shimakami, T., Hijikata, M., Luo, H., Ma, Y. Y., Kaneko, S., Shimotohno, K., Murakami, S. (2004). Effect of Interaction between Hepatitis C Virus NS5A and NS5B on Hepatitis C Virus RNA Replication with the Hepatitis C Virus Replicon. J. Virol. 78: 2738-2748 [Abstract] [Full Text]  
  • WARIS, G., SARKER, S., SIDDIQUI, A. (2004). Two-step affinity purification of the hepatitis C virus ribonucleoprotein complex. RNA 10: 321-329 [Abstract] [Full Text]  


Copyright © 2010 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»