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Characterization of nuclear localization of a hepatitis B virus precore protein derivative P22

Summary

Both of hepatitis B virus core protein and a precore protein derivative, named P22, have been shown to localize in the nucleus. Although P22 has ten additional amino acid residues at its amino-terminus, both proteins contain the same nuclear localization signal. In order to understand the mechanism that regulates the activity of this signal, we have studied the nuclear localization of P22 and compared it with that of core protein. It was found that both cytosolic and nuclear fractions of P22 were phosphorylated but to a lesser extent when compared with cytosolic core protein. This distinction was likely attributed to different conformations between these two proteins since the density gradient analysis revealed a different particle formation for P22 in the cytosol. When expressed in Vero cells synchronized by serum deprivation, P22 remained in the cytosol during G0 and G1 phases, accumulated gradually in the nucleus during S phase, and largely localized in the nucleus when cells were confluent. On the other hand, the core protein was transported into the nucleus during mid-G1 phase, shuttled back to the cytosol in S phase and again accumulated in the nucleus when cells were confluent. Interestingly, when aphidicolin was used to arrest the cells in late G1 phase, both proteins were found to accumulate in the nuclei. These results indicated that although both P22 and core proteins possessed the same nuclear localization signal, the cellular regulation of their nuclear transport was not identical and might involve different molecular mechanisms.

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References

  1. 1.

    Beasley RP, Trepo C, Stevens CE, Szmuness W (1977) The e antigen and vertical transmission of hepatitis B surface antigen. Am J Epidemiol 105: 94–98

  2. 2.

    Bruss V, Gerlich WH (1988) Formation of transmembrane hepatitis B e antigen by cotranslational in vitro processing of the viral precore protein. Virology 163: 268–275

  3. 3.

    Chu CM, Liaw YF (1987) Intrahepatic distribution of hepatitis B surface and core antigens in chronic hepatic B virus infection. Gastroenterology 92: 220–225

  4. 4.

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

  5. 5.

    Ganem D, Varmus HE (1987) The molecular biology of the heaptitis B viruses. Annu Rev Biochem 56: 651–693

  6. 6.

    Garcia PD, Ou JH, Rutter WJ, Walter P (1988) Targeting of the hepatitis B virus precore protein to the endoplasmic reticulum membrane: after signal peptide cleavage translocation can be aborted and the product released into the cytoplasm. J Cell Biol 106: 1093–1104

  7. 7.

    Graham FL, van der Ed AJ (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52: 456–467

  8. 8.

    Guidotti LG, Martinez V, Loh YT, Rogler CE, Chisari FV (1994) Hepatitis B virus nucleocapsid particles do not cross the hepatocyte nuclear membrane in transgenic mice. J Virol 60: 5469–5475

  9. 9.

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

  10. 10.

    Liao W, Ou JH (1995) Phosphorylation and nuclear localization of the hepatitis B virus core protein: significance of serine in the three repeated SPRRR motifs. J Virol 69: 1025–1029

  11. 11.

    Mondelli M, Tedder RS, Ferns B, Pontisso P, Realdi G, Alberti A (1986) Differential distribution of hepatitis B core and e antigens in hepatocytes: analysis by monoclonal antibodies. Hepatology 6: 199–204

  12. 12.

    Ou JH, Bell KD (1990) Comparative studies of hepatitis B virus precore and core particles. Virology 174: 185–191

  13. 13.

    Ou JH, Laub O, Rutter WJ (1986) Hepatitis B virus gene function: the precore region targets the core antigen to cellular membranes and causes the secretion of the e antigen. Proc Natl Acad Sci USA 83: 1578–1582

  14. 14.

    Ou JH, Standring DN, Masiarze FR, Rutter WJ (1988) A signal peptide encoded within the precore region of hepatitis B virus directs the secretion of a heterogeneous population of e antigens inXenopus oocytes. Proc Natl Acad Sci USA 85: 8405–8409

  15. 15.

    Ou JH, Yeh CT, Yen TSB (1989) Transport of hepatitis B virus precore protein into nucleus after cleavage of its signal peptide. J Virol 63: 5238–5243

  16. 16.

    Pedrali-Noy G, Spadari S, Miller-Faures A, Miller AOA, Kruppa J, Koch G (1980) Synchronization of Hela cell cultures by inhibition of DNA polymerase alpha with aphidicolin. Nucleic Acids Res 8: 377–387

  17. 17.

    Ponzetto A, Cote PJ, Ford EC, Purcell RH, Gerin JL (1984) Core antigen and antibody in woodchucks after infection with the woodchuck hepatitis virus. J Virol 52: 70–76

  18. 18.

    Robinson WS (1990) Hepadnaviridae and their replication. In: Fields BN, Knipe DM (eds) Virology. Raven Press, New York, pp 2137–2169

  19. 19.

    Roossnick MJ, Jameel S, Loukin SH, Siddiqui A (1986) Expression of hepatitis B virus core regions in mammalian cells. Mol Cell Biol 6: 1391–1400

  20. 20.

    Uy A, Vruss V, Gerlich WH, Kochel HG, Thomssen R (1986) Precore sequence of hepatitis B virus inducing e antigen and membrane association of the viral core protein, Virology 155: 89–96

  21. 21.

    Valenzuela P, Quiroga M, Zaldivar J, Gray P, Rutter WJ (1980) The nucleotide sequence of the hepatitis B viral genome and the identification of the major viral genes. In: Fields BN, Jaenisch R, Fox CF (ed) Animal virus genetics. Academic Press, New York, pp 57–70

  22. 22.

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

  23. 23.

    Yeh CT, Ou JH (1991) Phosphorylation of hepatitis B virus precore and core proteins. J Virol 65: 2327–2331

  24. 24.

    Yeh CT, Ou JH, Chu CM, Liaw YF (1994) Alteration of the subcellular localization of hepatitis B virus core protein by large but not small surface proteins. Biochem Biophys Res Commun 203: 1348–1354

  25. 25.

    Yeh CT, Wong SW, Fung YK, Ou JH (1993) Cell cycle regulation of nuclear localization of hepatitis B virus core protein. Proc Natl Acad Sci USA 90: 6459–6463

  26. 26.

    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: 1926–1932

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Yeh, C.-., Hong, L.-., Ou, J.-. et al. Characterization of nuclear localization of a hepatitis B virus precore protein derivative P22. Archives of Virology 141, 425–438 (1996). https://doi.org/10.1007/BF01718307

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Keywords

  • Cytosol
  • Nuclear Localization
  • Core Protein
  • Vero Cell
  • Nuclear Localization Signal