Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1988 Sep;62(9):3517–3521. doi: 10.1128/jvi.62.9.3517-3521.1988

DNA-binding activity of hepatitis B e antigen polypeptide lacking the protaminelike sequence of nucleocapsid protein of human hepatitis B virus.

K Matsuda 1, S Satoh 1, H Ohori 1
PMCID: PMC253481  PMID: 3404582

Abstract

The characteristics of binding of hepatitis B core antigen (HBcAg) and hepatitis B e antigen (HBeAg) polypeptides to hepatitis B virus (HBV) DNA were analyzed. HBcAg polypeptide from recombinant HBV core particles and HBeAg polypeptide from partially purified serum HBeAg were prepared and verified to have molecular weights of 21,500 (P21.5) and of 17,000 (P17) and 18,000 (P18), respectively, by immunoblot analysis. By reaction of these proteins on a nitrocellulose membrane with cloned 32P-HBV DNA, it was revealed that the HBeAg polypeptide, which lacks the C-terminal 34 amino acids of P21.5, as well as the HBcAg polypeptide, bound to the DNA. The secondary structures of nucleocapsid proteins of HBV, woodchuck hepatitis virus, and ground squirrel hepatitis virus were predicted by the Garnier algorithm. Amino acid sequences which, in addition to those of the C-terminal regions, may contribute to binding were proposed to be the 21-amino-acid residues located at amino acids 100 to 120 of the nucleocapsid proteins of these hepadnaviruses.

Full text

PDF
3517

Images in this article

3518Image
on p.3518
3518Image
on p.3518
3519Image
on p.3519

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bowen B., Steinberg J., Laemmli U. K., Weintraub H. The detection of DNA-binding proteins by protein blotting. Nucleic Acids Res. 1980 Jan 11;8(1):1–20. doi: 10.1093/nar/8.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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] [Google Scholar]
  3. Feitelson M. A., Marion P. L., Robinson W. S. Core particles of hepatitis B virus and ground squirrel hepatitis virus. II. Characterization of the protein kinase reaction associated with ground squirrel hepatitis virus and hepatitis B virus. J Virol. 1982 Aug;43(2):741–748. doi: 10.1128/jvi.43.2.741-748.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fujiyama A., Miyanohara A., Nozaki C., Yoneyama T., Ohtomo N., Matsubara K. Cloning and structural analyses of hepatitis B virus DNAs, subtype adr. Nucleic Acids Res. 1983 Jul 11;11(13):4601–4610. doi: 10.1093/nar/11.13.4601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Galibert F., Chen T. N., 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. doi: 10.1128/jvi.41.1.51-65.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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. doi: 10.1038/281646a0. [DOI] [PubMed] [Google Scholar]
  7. Garnier J., Osguthorpe D. J., Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. doi: 10.1016/0022-2836(78)90297-8. [DOI] [PubMed] [Google Scholar]
  8. Gerlich W. H., 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. doi: 10.1128/jvi.42.3.761-766.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hope I. A., Struhl K. Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast. Cell. 1986 Sep 12;46(6):885–894. doi: 10.1016/0092-8674(86)90070-x. [DOI] [PubMed] [Google Scholar]
  10. Kaplan P. M., Ford E. C., Purcell R. H., Gerin J. L. Demonstration of subpopulations of Dane particles. J Virol. 1976 Mar;17(3):885–893. doi: 10.1128/jvi.17.3.885-893.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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. doi: 10.1002/jmv.1890080404. [DOI] [PubMed] [Google Scholar]
  12. Magnius L. O., Espmark J. A. New specificities in Australia antigen positive sera distinct from the Le Bouvier determinants. J Immunol. 1972 Nov;109(5):1017–1021. [PubMed] [Google Scholar]
  13. Matsuda K., Kanno A., Ohori H. Immunochemical characterization of hepatitis B e antigen subtypes, HBeAg/1 and HBeAg/2, in sera of hepatitis B virus carriers. J Med Virol. 1986 Nov;20(3):219–228. doi: 10.1002/jmv.1890200304. [DOI] [PubMed] [Google Scholar]
  14. McLachlan A., Milich D. R., Raney A. K., Riggs M. G., Hughes J. L., Sorge J., Chisari F. V. Expression of hepatitis B virus surface and core antigens: influences of pre-S and precore sequences. J Virol. 1987 Mar;61(3):683–692. doi: 10.1128/jvi.61.3.683-692.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mishiro S., Imai M., Takahashi K., Machida A., Gotanda T., Miyakawa Y., Mayumi M. A 49,000-dalton polypeptide bearing all antigenic determinants and full immunogenicity of 22-nm hepatitis B surface antigen particles. J Immunol. 1980 Apr;124(4):1589–1593. [PubMed] [Google Scholar]
  16. Ohori H., Onodera S., Ishida N. Demonstration of hepatitis B e antigen (HBeAg) in association with intact Dane particles. J Gen Virol. 1979 May;43(2):423–427. doi: 10.1099/0022-1317-43-2-423. [DOI] [PubMed] [Google Scholar]
  17. 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. doi: 10.1099/0022-1317-65-2-405. [DOI] [PubMed] [Google Scholar]
  18. Ohori H., Yamada E., Tateda A., Ishida N. Prevalence of Williams e1 antigen in comparison with e2 antigen in hepatitis B antigen carriers and patients in hemodialysis unit. J Med Virol. 1980;6(1):61–74. doi: 10.1002/jmv.1890060109. [DOI] [PubMed] [Google Scholar]
  19. 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. doi: 10.1002/jmv.1890100209. [DOI] [PubMed] [Google Scholar]
  20. Ou J. H., Laub O., Rutter W. J. 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. doi: 10.1073/pnas.83.6.1578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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. doi: 10.1038/282575a0. [DOI] [PubMed] [Google Scholar]
  22. Petit M. A., 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. doi: 10.1128/jvi.53.2.543-551.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Roossinck M. J., Jameel S., Loukin S. H., Siddiqui A. Expression of hepatitis B viral core region in mammalian cells. Mol Cell Biol. 1986 May;6(5):1393–1400. doi: 10.1128/mcb.6.5.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roossinck M. J., Siddiqui A. In vivo phosphorylation and protein analysis of hepatitis B virus core antigen. J Virol. 1987 Apr;61(4):955–961. doi: 10.1128/jvi.61.4.955-961.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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] [Google Scholar]
  26. Seeger C., Ganem D., Varmus H. E. Nucleotide sequence of an infectious molecularly cloned genome of ground squirrel hepatitis virus. J Virol. 1984 Aug;51(2):367–375. doi: 10.1128/jvi.51.2.367-375.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Serrano M. A., Hirschman S. Z. Properties of hepatitis B e antigen synthesized by rat cells transfected with circular viral DNA. J Gen Virol. 1984 Aug;65(Pt 8):1373–1383. doi: 10.1099/0022-1317-65-8-1373. [DOI] [PubMed] [Google Scholar]
  28. 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] [Google Scholar]
  29. Takahashi K., Machida A., Funatsu G., Nomura M., Usuda S., Aoyagi S., Tachibana K., Miyamoto H., Imai M., Nakamura T. Immunochemical structure of hepatitis B e antigen in the serum. J Immunol. 1983 Jun;130(6):2903–2907. [PubMed] [Google Scholar]
  30. Tiollais P., Pourcel C., Dejean A. The hepatitis B virus. Nature. 1985 Oct 10;317(6037):489–495. doi: 10.1038/317489a0. [DOI] [PubMed] [Google Scholar]
  31. Vogt P. K., Bos T. J., Doolittle R. F. Homology between the DNA-binding domain of the GCN4 regulatory protein of yeast and the carboxyl-terminal region of a protein coded for by the oncogene jun. Proc Natl Acad Sci U S A. 1987 May;84(10):3316–3319. doi: 10.1073/pnas.84.10.3316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Will H., Cattaneo R., Pfaff E., Kuhn C., Roggendorf M., Schaller H. Expression of hepatitis B antigens with a simian virus 40 vector. J Virol. 1984 May;50(2):335–342. doi: 10.1128/jvi.50.2.335-342.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yamada E., Ishida N., Ohori H. Comparison of the antigenicity and protein composition of hepatitis B virus e antigen subtypes HBeAg/1 and HBeAg/2. J Gen Virol. 1983 Apr;64(Pt 4):895–903. doi: 10.1099/0022-1317-64-4-895. [DOI] [PubMed] [Google Scholar]
  34. Yamada E., Ohori H., Ishida N. Molecular heterogeneity of hepatitis B virus e antigen in liver and serum. J Gen Virol. 1981 Jul;55(Pt 1):75–86. doi: 10.1099/0022-1317-55-1-75. [DOI] [PubMed] [Google Scholar]
  35. 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. doi: 10.1016/0167-4838(82)90483-6. [DOI] [PubMed] [Google Scholar]
  36. Yoshizawa H., Itoh Y., Simonetti J. P., Takahashi T., Machida A., Miyakawa Y., Mayumi M. Demonstration of hepatitis B e antigen in hepatitis B core particles obtained from the nucleus of hepatocytes infected with hepatitis B virus. J Gen Virol. 1979 Mar;42(3):513–519. doi: 10.1099/0022-1317-42-3-513. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES