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J Virol. Feb 1985; 53(2): 543–551.
PMCID: PMC254669

HBc and HBe antigenicity and DNA-binding activity of major core protein P22 in hepatitis B virus core particles isolated from the cytoplasm of human liver cells.

Abstract

Highly purified hepatitis B virus core particles were obtained in large amounts from the cytoplasm of infected human liver cells. This DNA polymerase-negative core preparation had only hepatitis B core antigen-specific antigenicity and showed a surprising stability. Two forms of a single protein of 22,000 molecular weight, P22, were resolved electrophoretically; the slower moving species, P22a, appeared to be a reduced form of the protein, and the faster moving species, P22b, could have represented a conformational isomer containing an intramolecular disulfide bond(s). The immunological properties and DNA-binding activity of the reduced form, P22a, were examined following separation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by transfer onto nitrocellulose membranes (Western blotting). We found that the hepatitis B virus C gene protein shared the antigenic site responsible for both hepatitis B core and e antigen reactivity. We also demonstrated that the core protein(s) bound specifically the genomic hepatitis B virus DNA in comparison with a plasmid DNA (pBR322). This last observation was further substantiated by a radioimmunological method. P22a was also found to be phosphorylated in vitro by the endogenous protein kinase activity, copurified with the hepatitis B core antigen particles. These findings suggest that P22 is a multifunctional protein which is incorporated into core particles within the cytoplasm of the host cell before DNA encapsidation. A critical role of this protein in hepatitis B virus assembly is suggested.

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Selected References

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  • Alberti A, Pontisso P, Realdi G. Changes in hepatitis B virus DNA-polymerase activity in patients with chronic infection. J Med Virol. 1981;8(4):223–229. [PubMed]
  • Albin C, Robinson WS. Protein kinase activity in hepatitis B virus. J Virol. 1980 Apr;34(1):297–302. [PMC free article] [PubMed]
  • Barker LF, Almeida JD, Hoofnagle JH, Gerety RJ, Jackson DR, McGrath PP. Hepatitis B core antigen: immunology and electron microscopy. J Virol. 1974 Dec;14(6):1552–1558. [PMC free article] [PubMed]
  • Blobel G, Potter VR. Nuclei from rat liver: isolation method that combines purity with high yield. Science. 1966 Dec 30;154(3757):1662–1665. [PubMed]
  • Bowen B, Steinberg J, Laemmli UK, Weintraub H. The detection of DNA-binding proteins by protein blotting. Nucleic Acids Res. 1980 Jan 11;8(1):1–20. [PMC free article] [PubMed]
  • Budkowska A, Kalinowska B, Nowosławski A. Identification of two HBeAg subspecificities revealed by chemical treatment and enzymatic digestion of liver-derived HBcAg. J Immunol. 1979 Sep;123(3):1415–1416. [PubMed]
  • Budkowska A, Shih JW, Gerin JL. Immunochemistry and polypeptide composition of hepatitis B core antigen (HBc Ag). J Immunol. 1977 Apr;118(4):1300–1305. [PubMed]
  • Burnette WN. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. [PubMed]
  • Dane DS, Cameron CH, Briggs M. Virus-like particles in serum of patients with Australia-antigen-associated hepatitis. Lancet. 1970 Apr 4;1(7649):695–698. [PubMed]
  • Fraker PJ, Speck JC., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. [PubMed]
  • Gerlich WH, Goldmann U, Müller R, Stibbe W, Wolff W. Specificity and localization of the hepatitis B virus-associated protein kinase. J Virol. 1982 Jun;42(3):761–766. [PMC free article] [PubMed]
  • Gowans EJ, Burrell CJ, Jilbert AR, Marmion BP. Patterns of single- and double-stranded hepatitis B virus DNA and viral antigen accumulation in infected liver cells. J Gen Virol. 1983 Jun;64(Pt 6):1229–1239. [PubMed]
  • Hoofnagle JH, Gerety RJ, Barker LF. Antibody to hepatitis-B-virus core in man. Lancet. 1973 Oct 20;2(7834):869–873. [PubMed]
  • Hruska JF, Robinson WS. The proteins of hepatitis B Dane particle cores. J Med Virol. 1977;1(2):119–131. [PubMed]
  • Kaplan PM, Ford EC, Purcell RH, Gerin JL. Demonstration of subpopulations of Dane particles. J Virol. 1976 Mar;17(3):885–893. [PMC free article] [PubMed]
  • Kaplan PM, Greenman RL, Gerin JL, Purcell RH, Robinson WS. DNA polymerase associated with human hepatitis B antigen. J Virol. 1973 Nov;12(5):995–1005. [PMC free article] [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Ohori H, Shimizu N, Yamada E, Onodera S, Ishida N. Immunological and morphological properties of HBeAg subtypes (HBeAg/1 and HBeAg/2) in hepatitis B virus core particles. J Gen Virol. 1984 Feb;65(Pt 2):405–414. [PubMed]
  • Ohori H, Yamaki M, Onodera S, Yamada E, Ishida N. Antigenic conversion from HBcAg to HBeAg by degradation of hepatitis B core particles. Intervirology. 1980;13(2):74–82. [PubMed]
  • Onodera S, Ohori H, Yamaki M, Ishida N. Electron microscopy of human hepatitis B virus cores by negative staining-carbon film technique. J Med Virol. 1982;10(2):147–155. [PubMed]
  • Pasek M, Goto T, Gilbert W, Zink B, Schaller H, MacKay P, Leadbetter G, Murray K. Hepatitis B virus genes and their expression in E. coli. Nature. 1979 Dec 6;282(5739):575–579. [PubMed]
  • Peterson GL. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. [PubMed]
  • Pillot J, Petit MA. Immunochemical structure of the hepatitis B surface antigen vaccine--I. Treatment of immobilized HBsAg by dissociation agents with or without enzymatic digestion and identification of polypeptides by protein blotting. Mol Immunol. 1984 Jan;21(1):53–60. [PubMed]
  • Price P, Ostrove S, Flordellis C, Sells MA, Thung S, Gerber M, Christman J, Acs G. Characterization of RNA transcripts and virally coded proteins synthesized in mouse fibroblasts transfected with hepatitis B DNA: HBeAg synthesis in HBcAg-negative cells with active core-antigen genes. Biosci Rep. 1983 Nov;3(11):1017–1026. [PubMed]
  • Robinson WS, Clayton DA, Greenman RL. DNA of a human hepatitis B virus candidate. J Virol. 1974 Aug;14(2):384–391. [PMC free article] [PubMed]
  • Sakamoto Y, Yamada G, Mizuno M, Nishihara T, Kinoyama S, Kobayashi T, Takahashi T, Nagashima H. Full and empty particles of hepatitis B virus in hepatocytes from patients with HBsAg-positive chronic active hepatitis. Lab Invest. 1983 Jun;48(6):678–682. [PubMed]
  • Takahashi K, Akahane Y, Gotanda T, Mishiro T, Imai M, Miyakawa Y, Mayumi M. Demonstration of hepatitis B e antigen in the core of Dane particles. J Immunol. 1979 Jan;122(1):275–279. [PubMed]
  • Takahashi K, Imai M, Nomura M, Oinuma A, Machida A, Funatsu G, Miyakawa Y, Mayumi M. Demonstration of the immunogenicity of hepatitis B core antigen in a hepatitis B e antigen polypeptide (P19). J Gen Virol. 1981 Dec;57(Pt 2):325–330. [PubMed]
  • Takahashi K, Machida A, Funatsu G, Nomura M, Usuda S, Aoyagi S, Tachibana K, Miyamoto H, Imai M, Nakamura T, et al. Immunochemical structure of hepatitis B e antigen in the serum. J Immunol. 1983 Jun;130(6):2903–2907. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PMC free article] [PubMed]
  • Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed]

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