Gastroenterologia Japonica

, Volume 25, Supplement 2, pp 11–19 | Cite as

Hepadnavirus enhancer and its binding proteins

  • Seishi Murakami
  • Masato Uchijima
  • Atsushi Shimoda
  • Shuichi Kaneko
  • Kenichi Kobayashi
  • Nobu Hattori
Proceeding of the International Symposium on Biology of Hepatitis B Virus and Non-A Non-B Hepatitis Virus


In the hepadnavirus enhancer region, a 33 bp DNA sequence is strongly conserved among mammalian hepadnavirus genomes. To elucidate the role of the sequence, we tested enhancer activities and capability to form DNA-protein complex of several synthetic DNAs. Not only two tandem copies of a 46 bp DNA covering the sequence but also two tandem copies of a 23 bp in the sequence exhibit enhancer activity. Also the activity was augmented by treatment of a tumor promoter, TPA. DNA binding proteins complexes with the 23 bp DNA were augmented in extracts of HepG2 or HeLa cells stimulated with TPA. These results imply that the conserved sequence of hepadnavirus enhancer is a TPA-inducible enhancer which is transactivated by ubiquitous DNA-binding proteins. We presented results showing that DNA-protein complexes with a 23 bp DNA are similar to but distinct from those with a TPA-responsive element DNA, the recognition site for c-jun/fos products. We also presented results suggesting that hepadnavirus X protein may not directly or indirectly affect DNA-protein complex formation with the conserved sequence in the hepadnavirus enhancer.

Key words

DNA-binding protein enhancer hepadnavirus TPA-inducible 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Tiollais P, Pourcel C, Dejean A: The hepatitis B virus. Nature 1985;317:489–496PubMedCrossRefGoogle Scholar
  2. 2.
    Miller R, Keneko S, Chung CT, et al: Compact organization of hepatitis B virus genome. Hepatol 1989;9:322–327CrossRefGoogle Scholar
  3. 3.
    Shaul Y, Rutter WJ, Laub O: A human hepatitis B viral en-hancer element. EMBO J 1985;6:427–430Google Scholar
  4. 4.
    Tognoni P, Cattaneo R, Serfling E, et al: A novel expression se-lection approach allows precise mapping of the hepatitis B virus enhancer. Nucleic Acid Res 1985; 13:7457–7472PubMedCrossRefGoogle Scholar
  5. 5.
    Kodama K, Ogasawara N, Murakami S, et al: Nucleotide se-quence of a cloned woodchuck hepatits virus genome: Evolu-tional relationship between hepadnaviruses. J Virol 1985 ;56: 978–986PubMedGoogle Scholar
  6. 6.
    Dignam JD, Lebovity RM, Roeder RG: Accurate transcription initiation by RNA polymerase II in a soluble extract from iso-lated mammalian nuclei. Nucleic acid Res 1983;11:1475–1489PubMedCrossRefGoogle Scholar
  7. 7.
    Ide GJ, Sanuders CA: Rapid isolation of yeast nuclei. Current Genetics 1981;4:85–90CrossRefGoogle Scholar
  8. 8.
    Bradford, MM: A rapid and sensitive method for the quantita-tion of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 1976;72:248–254PubMedCrossRefGoogle Scholar
  9. 9.
    Chu G, Hayakawa H, Berg P: Electroporation for the efficient transfection of mammalian cells with DNA. Nucleic Acid Res 1987;15:1311–1326PubMedCrossRefGoogle Scholar
  10. 10.
    Gorman C, Moffat LF, Howard BH: Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 1982;2:1044–1051PubMedGoogle Scholar
  11. 11.
    Kaneko S, Oshima T, Murakami S, et al: Stable integration of woodchuck hepatitis virus DNA in transplanted tumors and established tissue culture cells derived from a woodchuck pri-mary hepatocellular carcinoma. Cancer Res 1986;46:3608–3613PubMedGoogle Scholar
  12. 12.
    Schleidereit C, Heguy A, Roeder RG: Identification and purifi-cation of a human lymphoid-specific octamer-binding protein (OTF2) that activates transcription of an immunoglobulin pro-moter in vitro. Cell 1987;51:783–793CrossRefGoogle Scholar
  13. 13.
    Laemmli UK: Cleavage of structural proteins during the assem-bly of the head of bacteriophage T4. Nature 1970;227:680–685PubMedCrossRefGoogle Scholar
  14. 14.
    Angel P, Allegretto EA, Okino, ST, et al: Oncogene jun en-codes a sequence-specific trans-activator similar to AP-1. Nature 1988;332:166–171PubMedCrossRefGoogle Scholar
  15. 15.
    Angel P, Imagawa M, Chiu, R, et al:cis element recognized by a TPA-modulatedtrans-acting factor. Cell 1987;49:729–739PubMedCrossRefGoogle Scholar
  16. 16.
    Lee W, Mitchell P, Tjian R: Purified transcription factor AP-1 interacts with TPA-inducible enhancer elements. Cell 1987;49: 741–752PubMedCrossRefGoogle Scholar
  17. 17.
    Rauscher III FJ, Smbucetti JC, Curran T, et al: Common DNA binding site for fos protein complexes and transcription factor AP-1. Cell 1988;52:471–480PubMedCrossRefGoogle Scholar
  18. 18.
    Twu J, Schloemer RH:Transcriptional trans-activating function of hepatitis B virus. J Virol 1987; 61:3448–3453PubMedGoogle Scholar
  19. 19.
    Seto E, Yen BTS, Peterlin BM, et al:Trans-activation of the hu-man immunodeficiency virus long terminal repeat by the hepa-titis B virus X protein. Proc Natl Acad Sci USA 1988;85:8286–9290PubMedCrossRefGoogle Scholar
  20. 20.
    Spandau DF, Lee C: trans-Activation of viral enhancers by the hepatitis B virus X protein. J Virol 1988;62:427–434PubMedGoogle Scholar
  21. 21.
    Twu J, Robinson WS; Hepatitis B virus X gene can transactivate heterologous virus sequences. Proc Natl Acad Sci USA 1989;86: 2046–2050PubMedCrossRefGoogle Scholar
  22. 22.
    Harshman Kd, Moye-Rowley WS, Parker CS: Transcriptional activation by the SV40 AP-1 recognition element in yeast is mediated by a factor similar to AP-1 that is distinct from GCN4. Cell 1988;53:321–330PubMedCrossRefGoogle Scholar
  23. 23.
    Jones RH, Moreno S, Nurse P, etal: Expression of the SV40 pro-moter in fission yeast:Identification and characterization an AP-1 like factor. Cell 1988;53:659–667PubMedCrossRefGoogle Scholar
  24. 24.
    Jones RH, Jones NC: Mammalian cAMP-responsive element can activate transcription in yeast and binds a yeast factor(s) that resembles the mammalian transcription factor ATF. Proc Natl Acad Sci USA 1989;86:2176–2180PubMedCrossRefGoogle Scholar
  25. 25.
    Yamamoto KK, Gonzalez GA, Biggs III WH, et al: Phos-phorylation-induced binding and transcriptional efficacy of nu-clear factor CRFB. Nature 1988;334:494–498PubMedCrossRefGoogle Scholar
  26. 26.
    Chiu R, Boyle WJ, Meek J, et al: The c-fos protein interacts with c-junlAP-1 to stimulate transcription of AP-1 responsive genes. Cell 1988;54:542–552CrossRefGoogle Scholar
  27. 27.
    Nakabeppu Y, Ryder K, Nathans D: DNA binding activities of three murine jun proteins: Stimulation by fos. Cell 1988;55:907–915PubMedCrossRefGoogle Scholar
  28. 28.
    Halazonetis TD, Georgopoulos K, Greenberg ME, et al: Jun dimerizes with itself and with c-fos forming complexes of dif-ferent DNA binding affinities. Cell 1988;55:917–924PubMedCrossRefGoogle Scholar
  29. 29.
    Rauscher III FJ, Cohen DR, Curran T, et al: fos-associated pro-tein p39 is the product of the jun proto-oncogene. Science 1988;240:1010–1016PubMedCrossRefGoogle Scholar
  30. 30.
    Wildeman AG, Zenke M, Schatz C, et al: Specific protein binding to the SV40 enhancer in vitro. Mol Cell Biol 1986;6:2098–2015PubMedGoogle Scholar
  31. 31.
    Elfassi E, Romet-Lemonne L, Essex M, et al: Evidence of extra-chromosomal forms of hepatitis B viral DNA in a bone marrow culture obtained from a patient recently infected with hepatitis B virus. Proc Natl Acad Asic USA 1984;81:3524–3528Google Scholar
  32. 32.
    Zelent AZ, Sells MA, Shvartsman M, et al: Replicative inter-mediate of hepatitis B virus in transfected murine fibroblasts. J Virol 1987;61:2921–2923PubMedGoogle Scholar
  33. 33.
    Vannice JJ, Levinson AD: Properties of the human hepatitis B virus enhancer: Position effects and cell-type nonspecificity. EMBO J 1988;6:1913–1920Google Scholar
  34. 34.
    Jameel SJ, Siddiqui A: The human hepatitis B virus enhancer re-quires trans-acting cellular factor(s) for activity. Mol Cell Biol 1986;6:710–715PubMedGoogle Scholar
  35. 35.
    Karpen S, Banerjee R, Zelent A, et al: Identification of protein-binding sites in the hepatitis B virus enhancer and core promoter domains. Mol Cell Biol 1988;8:5159–5165PubMedGoogle Scholar

Copyright information

© The Japanese Society of Gastroenlerology 1990

Authors and Affiliations

  • Seishi Murakami
    • 1
  • Masato Uchijima
    • 1
  • Atsushi Shimoda
    • 2
  • Shuichi Kaneko
    • 2
  • Kenichi Kobayashi
    • 2
  • Nobu Hattori
    • 2
  1. 1.Biophysics Department of Cancer Research InstituteKanazawa UniversityKanazawaJapan
  2. 2.First Department of Internal MedicineKanazawa University School of MedicineKanazawaJapan

Personalised recommendations