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A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein

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Hepatitis B Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1540))

Abstract

Hepatitis B virus (HBV) core protein (HBc) can be present in both nucleus and cytoplasm. The arginine-rich domain (ARD) at the cytoplasmic tail of HBc contains both a nuclear localization signal (NLS) and nuclear export signal (NES). We established a homokaryon assay to detect the dynamic trafficking of HBc between nucleus and cytoplasm in hepatocytes. Using immunofluorescence assay (IFA) and PEG-induced cell-cell fusion, we demonstrated that a chimeric reporter protein of SV40 large T antigen, when fused in-frame with HBc ARD, can shuttle from a donor nuclei (green) to the recipient nuclei (red) in the context of binucleated or polynucleated hybrid cells. The shuttling activity driven by HBc ARD can be measured quantitatively by this IFA method.

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References

  1. Shih C (2012) Chronic hepatitis B and C: basic science to clinical applications. World Scientific Publishing Co., NJ

    Book  Google Scholar 

  2. Chu CM, Yeh CT, Sheen IS, Liaw YF (1995) Subcellular localization of hepatitis B core antigen in relation to hepatocyte regeneration in chronic hepatitis B. Gastroenterology 109(6):1926–1932

    Article  CAS  PubMed  Google Scholar 

  3. Chu CM, Yeh CT, Chien RN, Sheen IS, Liaw YF (1997) The degrees of hepatocyte nuclear but not cytoplasmic expression of hepatitis B core antigen reflect the level of viral replication in chronic hepatitis B virus infection. J Clin Microbiol 35(1):102–105

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Kim TH, Cho EY, Oh HJ, Choi CS, Kim JW, Moon HB, Kim HC (2006) The degrees of hepatocyte cytoplasmic expression of hepatitis B core antigen correlate with histologic activity of liver disease in the young patients with chronic hepatitis B infection. J Korean Med Sci 21(2):279–283

    Article  PubMed  PubMed Central  Google Scholar 

  5. Naoumov NV, Portmann BC, Tedder RS, Ferns B, Eddleston AL, Alexander GJ, Williams R (1990) Detection of hepatitis B virus antigens in liver tissue. A relation to viral replication and histology in chronic hepatitis B infection. Gastroenterology 99(4):1248–1253

    Article  CAS  PubMed  Google Scholar 

  6. Liu CJ, Jeng YM, Chen CL, Cheng HR, Chen PJ, Chen TC, Liu CH, Lai MY, Chen DS, Kao JH (2009) Hepatitis B virus basal core promoter mutation and DNA load correlate with expression of hepatitis B core antigen in patients with chronic hepatitis B. J Infect Dis 199(5):742–749

    Article  CAS  PubMed  Google Scholar 

  7. Chu CM, Liaw YF (1987) Intrahepatic distribution of hepatitis B surface and core antigens in chronic hepatitis B virus infection. Hepatocyte with cytoplasmic/membranous hepatitis B core antigen as a possible target for immune hepatocytolysis. Gastroenterology 92(1):220–225

    Article  CAS  PubMed  Google Scholar 

  8. Hsu HC, Su IJ, Lai MY, Chen DS, Chang MH, Chuang SM, Sung JL (1987) Biologic and prognostic significance of hepatocyte hepatitis B core antigen expressions in the natural course of chronic hepatitis B virus infection. J Hepatol 5(1):45–50

    Article  CAS  PubMed  Google Scholar 

  9. Kalderon D, Roberts BL, Richardson WD, Smith AE (1984) A short amino acid sequence able to specify nuclear location. Cell 39(3 Pt 2):499–509

    Article  CAS  PubMed  Google Scholar 

  10. Yeh CT, Liaw YF, Ou JH (1990) The arginine-rich domain of hepatitis B virus precore and core proteins contains a signal for nuclear transport. J Virol 64(12):6141–6147

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Eckhardt SG, Milich DR, McLachlan A (1991) Hepatitis B virus core antigen has two nuclear localization sequences in the arginine-rich carboxyl terminus. J Virol 65(2):575–582

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Kann M, Sodeik B, Vlachou A, Gerlich WH, Helenius A (1999) Phosphorylation-dependent binding of hepatitis B virus core particles to the nuclear pore complex. J Cell Biol 145(1):45–55

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Li HC, Huang EY, Su PY, Wu SY, Yang CC, Lin YS, Chang WC, Shih C (2010) Nuclear export and import of human hepatitis B virus capsid protein and particles. PLoS Pathog 6(10):e1001162

    Article  PubMed  PubMed Central  Google Scholar 

  14. Yang CC, Huang EY, Li HC, Su PY, Shih C (2014) Nuclear export of human hepatitis B virus core protein and pregenomic RNA depends on the cellular NXF1-p15 machinery. PLoS One 9(10):e106683

    Article  PubMed  PubMed Central  Google Scholar 

  15. Wang S, Wang K, Zheng C (2013) Interspecies heterokaryon assay to characterize the nucleocytoplasmic shuttling of herpesviral proteins. Methods Mol Biol 1064:131–140

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was supported by Academia Sinica, National Health Research Institute (NHRI-EX102-10203BI), and Ministry of Science and Technology, Taiwan.

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Authors and Affiliations

  1. Taiwan International Graduate Program (TIGP) in Molecular Medicine, National Yang-Ming University and Academia Sinica, Taipei, Taiwan

    Ching-Chun Yang

  2. Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan

    Ching-Chun Yang, Hung-Cheng Li & Chiaho Shih

Authors
  1. Ching-Chun Yang
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  2. Hung-Cheng Li
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  3. Chiaho Shih
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Corresponding author

Correspondence to Chiaho Shih .

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Editors and Affiliations

  1. Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, Indiana, USA

    Haitao Guo

  2. Arbutus Biopharma, Inc., Doylestown, Pennsylvania, USA

    Andrea Cuconati

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Yang, CC., Li, HC., Shih, C. (2017). A Homokaryon Assay for Nucleocytoplasmic Shuttling Activity of HBV Core Protein. In: Guo, H., Cuconati, A. (eds) Hepatitis B Virus. Methods in Molecular Biology, vol 1540. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6700-1_5

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  • DOI: https://doi.org/10.1007/978-1-4939-6700-1_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6698-1

  • Online ISBN: 978-1-4939-6700-1

  • eBook Packages: Springer Protocols

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