This Article
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 Shih, C. M.
Right arrow Articles by Lee, Y. H.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shih, C. M.
Right arrow Articles by Lee, Y. H.

 Previous Article  |  Next Article 

J. Virol., Feb 1995, 1160-1171, Vol 69, No. 2
Copyright © 1995, American Society for Microbiology

Modulation of the trans-suppression activity of hepatitis C virus core protein by phosphorylation

CM Shih, CM Chen, SY Chen and YH Lee
Institute of Biochemistry, National Yang-Ming University, Taipei, Taiwan, Republic of China.

We previously demonstrated that the core protein of hepatitis C virus (HCV) can suppress gene expression and replication of hepatitis B virus (HBV) in a human hepatoma cell line (HuH-7). In this study, we have characterized the phosphorylation property of HCV core protein and examined the effect of phosphorylation on its suppressive activity of HBV. Our results indicated that both the full-length HCV core protein (22 kDa) and its processed or degraded forms (14 to 18 kDa) were phosphorylated in insect cells. As demonstrated by using the glutathione S-transferase fusion protein expression system and in vitro transcription and translation system, the phosphorylation of HCV core protein was carried out by protein kinase A (PKA) and protein kinase C (PKC) in vitro. In both kinase reactions, it was determined that the phosphorylated amino acid was a serine residue. The potential phosphorylated sites in core protein were identified as residues Ser-53 and Ser-116 for PKA and Ser-53 and Ser-99 for PKC. Comparison of the phosphorylation intensities of the wild type and Ser mutants suggested that Ser-99 and Ser-116 were the major phosphorylation sites for PKC and PKA, respectively. The phosphorylation of Ser-99 and Ser-116, but not Ser-53, in HCV core protein was essential for the suppressive activity of HCV core protein on HBV gene expression and replication in HuH-7 cells. Mutation of the former two serine residues to alanine or aspartate residues led to a drastic loss of the inhibitory effects of HCV core protein on HBV gene expression (both transcription and antigen production) and pregenomic RNA encapsidation, as well as the release of HBV virus particles. In contrast, the Ser-53 mutant conferred the same level of suppressive activity as the wild type did. This property is in accordance with the observation that Ser-99 and Ser-116 are the predominant phosphorylation sites in the HCV core construct. All serine mutants (including those with mutations in PKA, PKC, and both kinase recognition sites) of HCV core protein retained the ability to translocate into the nucleus. Furthermore, wild-type HCV core protein diminished its suppressive activity when cells were treated with PKA or PKC inhibitor. In conclusion, HCV core protein is a phospho-protein and in HuH-7 cells, its trans suppression of HBV gene expression and replication is positively regulated by PKA and PKC. The role of phosphorylation in the control of trans-suppressive activity cannot be reproduced by introducing an acidic residue.(ABSTRACT TRUNCATED AT 250 WORDS)


This article has been cited by other articles:

  • Murray, C. L., Jones, C. T., Tassello, J., Rice, C. M. (2007). Alanine Scanning of the Hepatitis C Virus Core Protein Reveals Numerous Residues Essential for Production of Infectious Virus. J. Virol. 81: 10220-10231 [Abstract] [Full Text]  
  • Guo, H., Zhou, T., Jiang, D., Cuconati, A., Xiao, G.-H., Block, T. M., Guo, J.-T. (2007). Regulation of Hepatitis B Virus Replication by the Phosphatidylinositol 3-Kinase-Akt Signal Transduction Pathway. J. Virol. 81: 10072-10080 [Abstract] [Full Text]  
  • Rodriguez-Inigo, E., Bartolome, J., Ortiz-Movilla, N., Platero, C., Lopez-Alcorocho, J. M., Pardo, M., Castillo, I., Carreno, V. (2005). Hepatitis C Virus (HCV) and Hepatitis B Virus (HBV) Can Coinfect the Same Hepatocyte in the Liver of Patients with Chronic HCV and Occult HBV Infection. J. Virol. 79: 15578-15581 [Abstract] [Full Text]  
  • Joo, M., Hahn, Y. S., Kwon, M., Sadikot, R. T., Blackwell, T. S., Christman, J. W. (2005). Hepatitis C Virus Core Protein Suppresses NF-{kappa}B Activation and Cyclooxygenase-2 Expression by Direct Interaction with I{kappa}B Kinase {beta}. J. Virol. 79: 7648-7657 [Abstract] [Full Text]  
  • Klein, K. C., Dellos, S. R., Lingappa, J. R. (2005). Identification of Residues in the Hepatitis C Virus Core Protein That Are Critical for Capsid Assembly in a Cell-Free System. J. Virol. 79: 6814-6826 [Abstract] [Full Text]  
  • Guix, S., Caballero, S., Bosch, A., Pinto, R. M. (2004). C-Terminal nsP1a Protein of Human Astrovirus Colocalizes with the Endoplasmic Reticulum and Viral RNA. J. Virol. 78: 13627-13636 [Abstract] [Full Text]  
  • Ogino, T., Fukuda, H., Imajoh-Ohmi, S., Kohara, M., Nomoto, A. (2004). Membrane Binding Properties and Terminal Residues of the Mature Hepatitis C Virus Capsid Protein in Insect Cells. J. Virol. 78: 11766-11777 [Abstract] [Full Text]  
  • Maillard, P., Lavergne, J.-P., Siberil, S., Faure, G., Roohvand, F., Petres, S., Teillaud, J. L., Budkowska, A. (2004). Fc{gamma} Receptor-like Activity of Hepatitis C Virus Core Protein. J. Biol. Chem. 279: 2430-2437 [Abstract] [Full Text]  
  • Chen, S.-Y., Kao, C.-F., Chen, C.-M., Shih, C.-M., Hsu, M.-J., Chao, C.-H., Wang, S.-H., You, L.-R., Lee, Y.-H. W. (2003). Mechanisms for Inhibition of Hepatitis B Virus Gene Expression and Replication by Hepatitis C Virus Core Protein. J. Biol. Chem. 278: 591-607 [Abstract] [Full Text]  
  • Ogata, S., Nagano-Fujii, M., Ku, Y., Yoon, S., Hotta, H. (2002). Comparative Sequence Analysis of the Core Protein and Its Frameshift Product, the F Protein, of Hepatitis C Virus Subtype 1b Strains Obtained from Patients with and without Hepatocellular Carcinoma. J. Clin. Microbiol. 40: 3625-3630 [Abstract] [Full Text]  
  • Wootton, S. K., Rowland, R. R. R., Yoo, D. (2002). Phosphorylation of the Porcine Reproductive and Respiratory Syndrome Virus Nucleocapsid Protein. J. Virol. 76: 10569-10576 [Abstract] [Full Text]  
  • Lee, M. N., Jung, E. Y., Kwun, H. J., Jun, H. K., Yu, D.-Y., Choi, Y. H., Jang, K. L. (2002). Hepatitis C virus core protein represses the p21 promoter through inhibition of a TGF-{beta} pathway. J. Gen. Virol. 83: 2145-2151 [Abstract] [Full Text]  
  • Lu, W., Strohecker, A., Ou, J.-h. (2001). Post-translational Modification of the Hepatitis C Virus Core Protein by Tissue Transglutaminase. J. Biol. Chem. 276: 47993-47999 [Abstract] [Full Text]  
  • Bergqvist, A., Rice, C. M. (2001). Transcriptional Activation of the Interleukin-2 Promoter by Hepatitis C Virus Core Protein. J. Virol. 75: 772-781 [Abstract] [Full Text]  
  • Aoki, H., Hayashi, J., Moriyama, M., Arakawa, Y., Hino, O. (2000). Hepatitis C Virus Core Protein Interacts with 14-3-3 Protein and Activates the Kinase Raf-1. J. Virol. 74: 1736-1741 [Abstract] [Full Text]  
  • You, L.-R., Chen, C.-M., Yeh, T.-S., Tsai, T.-Y., Mai, R.-T., Lin, C.-H., Lee, Y.-H. W. (1999). Hepatitis C Virus Core Protein Interacts with Cellular Putative RNA Helicase. J. Virol. 73: 2841-2853 [Abstract] [Full Text]  
  • You, L.-R., Chen, C.-M., Lee, Y.-H. W. (1999). Hepatitis C Virus Core Protein Enhances NF-kappa B Signal Pathway Triggering by Lymphotoxin-beta Receptor Ligand and Tumor Necrosis Factor Alpha. J. Virol. 73: 1672-1681 [Abstract] [Full Text]  
  • Hsieh, T.-Y., Matsumoto, M., Chou, H.-C., Schneider, R., Hwang, S. B., Lee, A. S., Lai, M. M. C. (1998). Hepatitis C Virus Core Protein Interacts with Heterogeneous Nuclear Ribonucleoprotein K. J. Biol. Chem. 273: 17651-17659 [Abstract] [Full Text]  
  • Zhu, N., Khoshnan, A., Schneider, R., Matsumoto, M., Dennert, G., Ware, C., Lai, M. M. C. (1998). Hepatitis C Virus Core Protein Binds to the Cytoplasmic Domain of Tumor Necrosis Factor (TNF) Receptor 1 and Enhances TNF-Induced Apoptosis. J. Virol. 72: 3691-3697 [Abstract] [Full Text]  
  • Ray, R. B., Steele, R., Meyer, K., Ray, R. (1997). Transcriptional Repression of p53 Promoter by Hepatitis C Virus Core Protein. J. Biol. Chem. 272: 10983-10986 [Abstract] [Full Text]  
  • Otsuka, M., Kato, N., Lan, K.-H., Yoshida, H., Kato, J., Goto, T., Shiratori, Y., Omata, M. (2000). Hepatitis C Virus Core Protein Enhances p53 Function through Augmentation of DNA Binding Affinity and Transcriptional Ability. J. Biol. Chem. 275: 34122-34130 [Abstract] [Full Text]  
  • Huang, W.-H., Yung, B. Y. M., Syu, W.-J., Lee, Y.-H. W. (2001). The Nucleolar Phosphoprotein B23 Interacts with Hepatitis Delta Antigens and Modulates the Hepatitis Delta Virus RNA Replication. J. Biol. Chem. 276: 25166-25175 [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»