Serving the Society Since 1986
Advanced Search
Volume 25 Issue 5
October 2010
    Citation: Yuan-peng ZHANG, Rong-wu ZHANG, Wei-shan CHANG, Yan-yan WANG. Cxcl16 Interact With SARS-CoV N Protein In and Out Cell .VIROLOGICA SINICA, 2010, 25(5) : 369-374.  http://dx.doi.org/10.1007/s12250-010-3129-x
    shu

    Cxcl16 Interact With SARS-CoV N Protein In and Out Cell

    cstr: 32224.14.s12250-010-3129-x
    • 1.

      College of Animal Science and Technology, Shandong Agricultural University, Tai′an 271018, China

    • 2.

      Ling County Animal Husboundry Bureall, Dezhou 253500, China

    • Corresponding author: Wei-shan CHANG, wschang@sdau.edu.cn
    • Received Date: 22 January 2010
      Accepted Date: 30 June 2010
      Available online: 01 October 2010
    • Our study investigated the host cell protein which can interact with SARS-CoV N protein, and explored the functional connections. The eukaryotic expression vectors pEGFP-N1/SARS-CoVN and pdsRed2-N1/ CXCL16 were constructed and used to co-transfect HEK293FT cells by the calcium phosphate method. The HIS-tagged fusion protein SARS-CoVN-GFP was then built and purified for the binding assay in vitro. The co-localization of SARS-CoVN and CXCL16 in the cytoplasm of HEK293FT cells was also shown using confocal laser scanning microscopy. It is suggested that their interaction might be through direct combination. Under a fluorescence microscope, it was observed that the purified fusion protein SARS-CoVN-GFP was attached to the cell membrane of CXCL16-transfected cells, indicating that SARS-CoVN and CXCL16 can be mutually combined.
    • 加载中

      1. Brunn A A, Teepe C, Simpson J C, et al. 2007. Analysis of intraviral protein-protein interactions of the SARS coronavirus ORFeome. PLoS ONE, 23: 459.

      2. Chang W S, Zhai J, Song W G. 2008. Screen and Identification of The Protein-protein Interactors in The HostCell With The SARS Coronavirus Nucleocapsid Protein. Prog Biochem Biophy, 35 (9): 1007-1013. (in Chinese)

      3. Fan Z, Zhou Y, Tan X, et al. 2006. SARS-CoV nucleocapsid protein binds to hUbc9, a ubiquitin conjugating enzyme of the sumoylation system. J Med Virol, 78: 1365-1373.
          doi: 10.1002/(ISSN)1096-9071

      4. Kopecky-Bromberg S A, Martínez-Sobrido L, Frieman M, et al. 2007. Severe acute respiratory syndrome coronavirus open reading frame (ORF) 3b, ORF 6, and nucleocapsid proteins function as interferon antagonists. J Virol, 81: 548-557.
          doi: 10.1128/JVI.01782-06

      5. Lalani A S, Graham K, Mossman K, et al. 1997. The purified myxoma virus gamma interferon receptor homolog M-T7 interacts with the heparin domains of chemokines. J Virol, 71 (6): 4356-4363.

      6. Luo C, Luo H, Zheng S, et al. 2004. Nucleocapsid protein of SARS coronavirus tightly binds to human cyclophilin A, Biochem Biophys Res Commun, 321: 557-565.
          doi: 10.1016/j.bbrc.2004.07.003

      7. Luo H, Chen Q, Chen J, et al. 2005. The nucleocapsid protein of SARS coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleo-protein A1, FEBS letters, 579: 2623-2628.
          doi: 10.1016/j.febslet.2005.03.080

      8. Matloubian M, David A, Engel S, et al. 2000.A transmembrane CXC chemokine is a ligand for HIV-coreceptor Bonzo. Nat Immunol, 1 (4): 298-304.
          doi: 10.1038/79738

      9. Saikatendu K S, Jeremiah J S, Subramanian V, et al. 2007. Ribonucleocapsid Formation of Severe Acute Respiratory Syndrome Coronavirus through Molecular Action of the N-Terminal Domain of N Protein. J Virol, 81: 3913-3921.
          doi: 10.1128/JVI.02236-06

      10. Shashkin P, Simpson D, Mishin V, et al.2003. Expression of CXCL16 in human T cells. Arterioscler Thromb Vasc Biol, 23 (1): 148-149.
          doi: 10.1161/01.ATV.0000043906.61088.4B

      11. Surjit M, Kumar R, Mishra R N, et al. 2005. The Severe Acute Respiratory Syndrome Coronavirus Nucleocapsid Protein Is Phosphorylated and Localizes in the Cytoplasm by 14-3-3-Mediated Translocation, J Virol, 79: 11476-11486.
          doi: 10.1128/JVI.79.17.11476-11486.2005

      12. Surjit M, Liu B, Chow V T, et al. 2006. The Nucleocapsid Protein of Severe Acute Respiratory Syndrome-Coronavirus Inhibits the Activity of Cyclin-Cyclin-dependent Kinase Complex and Blocks S Phase Progression in Mammalian Cells. J Biol Chem, 281: 10669-10681.
          doi: 10.1074/jbc.M509233200

      13. Wei W J, Wen H Y. 2007. The role of Chemokine CXCL16 in clinic diseases. Prog Modern Biomed, 7 (5): 798-800. (in Chinese).

      14. Wilbanks A, Zondlo S C, Murphy K, et al. 2001. Expression cloning of the STRL33/BONZO/TYMSTR ligand reveals elements of CC, CXC, and CX3C chemokines. J Immunol, 166 (8): 5145-5154.
          doi: 10.4049/jimmunol.166.8.5145

      15. Zhao X G, Nicholls J M, Chen Y G. 2007. Sars-cov nucleocapsid protein interacts with smad3 and modulates TGF-β signaling. J Biol Chem, 283: 3272-3280.

    • 加载中

    Figures(5)

    PDF

    Article Metrics

    Article views(4146) PDF downloads(11) Cited by()

    Related
    Proportional views

      Cxcl16 Interact With SARS-CoV N Protein In and Out Cell

        Corresponding author: Wei-shan CHANG, wschang@sdau.edu.cn
      • 1. College of Animal Science and Technology, Shandong Agricultural University, Tai′an 271018, China
      • 2. Ling County Animal Husboundry Bureall, Dezhou 253500, China
      • Received Date: 22 January 2010
      • Accepted Date: 30 June 2010

      Abstract: Our study investigated the host cell protein which can interact with SARS-CoV N protein, and explored the functional connections. The eukaryotic expression vectors pEGFP-N1/SARS-CoVN and pdsRed2-N1/ CXCL16 were constructed and used to co-transfect HEK293FT cells by the calcium phosphate method. The HIS-tagged fusion protein SARS-CoVN-GFP was then built and purified for the binding assay in vitro. The co-localization of SARS-CoVN and CXCL16 in the cytoplasm of HEK293FT cells was also shown using confocal laser scanning microscopy. It is suggested that their interaction might be through direct combination. Under a fluorescence microscope, it was observed that the purified fusion protein SARS-CoVN-GFP was attached to the cell membrane of CXCL16-transfected cells, indicating that SARS-CoVN and CXCL16 can be mutually combined.

        HTML