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J Virol. 1989 Nov; 63(11): 4645–4652.
PMCID: PMC251098

A recombinant hepatitis B core antigen polypeptide with the protamine-like domain deleted self-assembles into capsid particles but fails to bind nucleic acids.


We have cloned in Escherichia coli both the complete core gene of hepatitis B virus and a truncated version of it, leading to the synthesis of high levels of a core-antigen-equivalent polypeptide (r-p22) and of an e-antigen-equivalent polypeptide (r-p16), respectively. We then compared the structural and antigenic properties of the two polypeptides, as well as their ability to bind viral nucleic acids. r-p16 was found to self-assemble into capsid-like particles that appeared similar, when observed under the electron microscope, to those formed by r-p22. In r-p16 particles, disulfide bonds linked the truncated polypeptides in dimers, assembled in the particle by noncovalent interactions. In r-p22 capsids, further disulfide bonds, conceivably involving the carboxy-terminal cysteines of r-p22 polypeptides, joined the dimers together, converting the structure into a covalently closed lattice. The protamine-like domain was at least partly exposed on the surface of r-p22 particles, since it was accessible to selective proteolysis. Finally, r-p22, but not r-p16, was shown to bind native and denatured DNA as well as RNA. Taken together, these results suggest that the protamine-like domain in core polypeptides is a nucleic acid-binding domain and is dispensable for the correct folding and assembly of amino-terminal and central regions.

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Selected References

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  • Argos P, Fuller SD. A model for the hepatitis B virus core protein: prediction of antigenic sites and relationship to RNA virus capsid proteins. EMBO J. 1988 Mar;7(3):819–824. [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]
  • Bruss V, Gerlich WH. Formation of transmembraneous hepatitis B e-antigen by cotranslational in vitro processing of the viral precore protein. Virology. 1988 Apr;163(2):268–275. [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]
  • Burrell CJ, Mackay P, Greenaway PJ, Hofschneider PH, Murray K. Expression in Escherichia coli of hepatitis B virus DNA sequences cloned in plasmid pBR322. Nature. 1979 May 3;279(5708):43–47. [PubMed]
  • Cabilly S, Riggs AD, Pande H, Shively JE, Holmes WE, Rey M, Perry LJ, Wetzel R, Heyneker HL. Generation of antibody activity from immunoglobulin polypeptide chains produced in Escherichia coli. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3273–3277. [PMC free article] [PubMed]
  • Chang C, Enders G, Sprengel R, Peters N, Varmus HE, Ganem D. Expression of the precore region of an avian hepatitis B virus is not required for viral replication. J Virol. 1987 Oct;61(10):3322–3325. [PMC free article] [PubMed]
  • Cohen BJ, Richmond JE. Electron microscopy of hepatitis B core antigen synthesized in E. coli. Nature. 1982 Apr 15;296(5858):677–679. [PubMed]
  • de Boer HA, Comstock LJ, Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. [PMC free article] [PubMed]
  • Edman JC, Hallewell RA, Valenzuela P, Goodman HM, Rutter WJ. Synthesis of hepatitis B surface and core antigens in E. coli. Nature. 1981 Jun 11;291(5815):503–506. [PubMed]
  • Ferns RB, Tedder RS. Monoclonal antibodies to hepatitis Be antigen (HBeAg) derived from hepatitis B core antigen (HBcAg): their use in characterization and detection of HBeAg. J Gen Virol. 1984 May;65(Pt 5):899–908. [PubMed]
  • Galibert F, Chen TN, Mandart E. Nucleotide sequence of a cloned woodchuck hepatitis virus genome: comparison with the hepatitis B virus sequence. J Virol. 1982 Jan;41(1):51–65. [PMC free article] [PubMed]
  • Galibert F, Mandart E, Fitoussi F, Tiollais P, Charnay P. Nucleotide sequence of the hepatitis B virus genome (subtype ayw) cloned in E. coli. Nature. 1979 Oct 25;281(5733):646–650. [PubMed]
  • Gerber MA, Sells MA, Chen ML, Thung SN, Tabibzadeh SS, Hood A, Acs G. Morphologic, immunohistochemical, and ultrastructural studies of the production of hepatitis B virus in vitro. Lab Invest. 1988 Aug;59(2):173–180. [PubMed]
  • Gershoni JM. Protein blotting: a manual. Methods Biochem Anal. 1988;33:1–58. [PubMed]
  • Imai M, Nomura M, Gotanda T, Sano T, Tachibana K, Miyamoto H, Takahashi K, Toyama S, Miyakawa Y, Mayumi M. Demonstration of two distinct antigenic determinants on hepatitis B e antigen by monoclonal antibodies. J Immunol. 1982 Jan;128(1):69–72. [PubMed]
  • Kobayashi M, Koike K. Complete nucleotide sequence of hepatitis B virus DNA of subtype adr and its conserved gene organization. Gene. 1984 Oct;30(1-3):227–232. [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]
  • MacKay P, Lees J, Murray K. The conversion of hepatitis B core antigen synthesized in E coli into e antigen. J Med Virol. 1981;8(4):237–243. [PubMed]
  • Magnius LO, Espmark JA. New specificities in Australia antigen positive sera distinct from the Le Bouvier determinants. J Immunol. 1972 Nov;109(5):1017–1021. [PubMed]
  • Matsuda K, Ohori H. Immunochemical characteristics of hepatitis B e antigen subspecificities, HBeAg/1 and HBeAg/2. J Immunol. 1988 Sep 1;141(5):1709–1713. [PubMed]
  • Matsuda K, Satoh S, Ohori H. DNA-binding activity of hepatitis B e antigen polypeptide lacking the protaminelike sequence of nucleocapsid protein of human hepatitis B virus. J Virol. 1988 Sep;62(9):3517–3521. [PMC free article] [PubMed]
  • McLachlan A, Milich DR, Raney AK, Riggs MG, Hughes JL, Sorge J, Chisari FV. Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences. J Virol. 1987 Mar;61(3):683–692. [PMC free article] [PubMed]
  • Moriarty AM, Hoyer BH, Shih JW, Gerin JL, Hamer DH. Expression of the hepatitis B virus surface antigen gene in cell culture by using a simian virus 40 vector. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2606–2610. [PMC free article] [PubMed]
  • Nassal M. Total chemical synthesis of a gene for hepatitis B virus core protein and its functional characterization. Gene. 1988 Jun 30;66(2):279–294. [PubMed]
  • Neurath AR, Strick N, Szmuness W, Stevens CE, Harley EJ. Radioimmunoassay of hepatitis B e-antigen (HBeAg): identification of HBeAg not associated with immunoglobulins. J Gen Virol. 1979 Mar;42(3):493–504. [PubMed]
  • Ono Y, Onda H, Sasada R, Igarashi K, Sugino Y, Nishioka K. The complete nucleotide sequences of the cloned hepatitis B virus DNA; subtype adr and adw. Nucleic Acids Res. 1983 Mar 25;11(6):1747–1757. [PMC free article] [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]
  • Ou JH, Laub O, Rutter WJ. 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 U S A. 1986 Mar;83(6):1578–1582. [PMC free article] [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]
  • Petit MA, Pillot J. 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. J Virol. 1985 Feb;53(2):543–551. [PMC free article] [PubMed]
  • Robbins SG, Frana MF, McGowan JJ, Boyle JF, Holmes KV. RNA-binding proteins of coronavirus MHV: detection of monomeric and multimeric N protein with an RNA overlay-protein blot assay. Virology. 1986 Apr 30;150(2):402–410. [PubMed]
  • Roossinck MJ, Jameel S, Loukin SH, Siddiqui A. Expression of hepatitis B viral core region in mammalian cells. Mol Cell Biol. 1986 May;6(5):1393–1400. [PMC free article] [PubMed]
  • Salfeld J, Pfaff E, Noah M, Schaller H. Antigenic determinants and functional domains in core antigen and e antigen from hepatitis B virus. J Virol. 1989 Feb;63(2):798–808. [PMC free article] [PubMed]
  • Schlicht HJ, Bartenschlager R, Schaller H. The duck hepatitis B virus core protein contains a highly phosphorylated C terminus that is essential for replication but not for RNA packaging. J Virol. 1989 Jul;63(7):2995–3000. [PMC free article] [PubMed]
  • Schlicht HJ, Salfeld J, Schaller H. The duck hepatitis B virus pre-C region encodes a signal sequence which is essential for synthesis and secretion of processed core proteins but not for virus formation. J Virol. 1987 Dec;61(12):3701–3709. [PMC free article] [PubMed]
  • Seeger C, Ganem D, Varmus HE. Nucleotide sequence of an infectious molecularly cloned genome of ground squirrel hepatitis virus. J Virol. 1984 Aug;51(2):367–375. [PMC free article] [PubMed]
  • Standring DN, Ou JH, Masiarz FR, Rutter WJ. A signal peptide encoded within the precore region of hepatitis B virus directs the secretion of a heterogeneous population of e antigens in Xenopus oocytes. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8405–8409. [PMC free article] [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]
  • Tedder RS, Bull FG. Characterization of 'e' antigen associated with hepatitis B. Clin Exp Immunol. 1979 Mar;35(3):380–389. [PMC free article] [PubMed]
  • Uy A, Bruss V, Gerlich WH, Köchel HG, Thomssen R. Precore sequence of hepatitis B virus inducing e antigen and membrane association of the viral core protein. Virology. 1986 Nov;155(1):89–96. [PubMed]
  • Vaudin M, Wolstenholme AJ, Tsiquaye KN, Zuckerman AJ, Harrison TJ. The complete nucleotide sequence of the genome of a hepatitis B virus isolated from a naturally infected chimpanzee. J Gen Virol. 1988 Jun;69(Pt 6):1383–1389. [PubMed]
  • Weimer T, Salfeld J, Will H. Expression of the hepatitis B virus core gene in vitro and in vivo. J Virol. 1987 Oct;61(10):3109–3113. [PMC free article] [PubMed]
  • Yamaki M, Ohori H, Onodera S, Ishida N, Maeda H. Circular dichroism and biochemical properties of the hepatitis B virus core antigen. Biochim Biophys Acta. 1982 Sep 7;706(2):165–173. [PubMed]

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