1887

Abstract

Hepatitis B virus (HBV) has a double-stranded DNA genome. The minus-strand contains coding regions for all known HBV proteins and most of the -regulatory elements. Little is known about transcription from the S-(+)-strand and its regulation. Thus, the presence of regulatory elements located on the S-(+)-strand was investigated by inserting nt 1038–1783 of HBV in both orientations between the human cytomegalovirus (HCMV) promoter and a luciferase gene. Transfection experiments revealed that the plasmid containing this HBV DNA fragment in an orientation allowing expression from the S-(+)-strand (antisense) led to inhibition of luciferase gene expression compared to the plasmid containing this sequence in an orientation that allows gene expression from the L-(−)-strand (sense). Deletion analyses delimit the sequence essential for the inhibitory effect to a 150 bp region that also carries part of the enhancerII/core promoter complex. However, the possible influence of this regulatory element has been excluded in various experiments. The repressing HBV sequence acts in an orientation- and position-dependent manner; no inhibition was observed when this DNA element was inserted upstream of the HCMV promoter or downstream of the luciferase gene. Northern blot analyses revealed reduced luciferase mRNA steady-state levels in cells transfected with constructs containing the essential HBV sequence in antisense orientation compared to plasmids containing this sequence in sense orientation. Since nuclear run-on experiments showed similar transcription initiation rates with these plasmids, the diminished luciferase mRNA steady-state levels must be due to altered stabilities, suggesting that nt 1783–1638 of HBV encode an RNA-destabilizing element.

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1999-10-01
2020-01-22
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References

  1. Alonso, S. , Minty, A. , Bourlet, Y. & Buckingham, M. ( 1986; ). Comparison of three actin-coding sequences in the mouse; evolutionary relationships between the actin genes of warm-blooded vertebrates. Journal of Molecular Evolution 23, 11-22.[CrossRef]
    [Google Scholar]
  2. Alt, M. , Renz, R. , Hofschneider, P. H. , Paumgartner, G. & Caselmann, W. H. ( 1995; ). Specific inhibition of hepatitis C viral gene expression by antisense phosphorothioate oligodeoxynucleotides. Hepatology 22, 707-717.
    [Google Scholar]
  3. Bellman, C. A. & Parker, R. ( 1995; ). Degradation of mRNA in eukaryotes. Cell 81, 179-183.[CrossRef]
    [Google Scholar]
  4. Brand, A. H. , Breeden, L. , Abraham, J. , Sternglanz, R. & Nasmyth, K. ( 1985; ). Characterization of a ‘silencer’ in yeast: a DNA sequence with properties opposite to those of a transcriptional enhancer. Cell 41, 41-48.[CrossRef]
    [Google Scholar]
  5. Chen, M. & Ou, J.-H. ( 1995; ). Cell type-dependent regulation of the activity of the negative regulatory element of the hepatitis B virus core promoter. Virology 214, 198-206.[CrossRef]
    [Google Scholar]
  6. Chen, C.-Y. A. & Shyu, A.-B. ( 1995; ). AU-rich elements: characterization and importance in mRNA degradation. Trends in Biochemical Sciences 20, 465-470.[CrossRef]
    [Google Scholar]
  7. Chen, I.-H. , Huang, C.-J. & Ting, L.-P. ( 1995; ). Overlapping initiator and TATA box functions in the basal core promoter of hepatitis B virus. Journal of Virology 69, 3647-3657 .
    [Google Scholar]
  8. Donello, J. E. , Beeche, A. A. , Smith, G. J.III , Lucero, G. R. & Hope, T. J. ( 1996; ). The hepatitis B virus posttranscriptional regulatory element is composed of two subelements. Journal of Virology 70, 4345-4351 .
    [Google Scholar]
  9. Donello, J. E. , Loeb, J. E. & Hope, T. J. ( 1998; ). Woodchuck hepatitis virus contains a tripartite posttranscriptional regulatory element. Journal of Virology 72, 5085-5092 .
    [Google Scholar]
  10. Farrell, R. E., Jr (1993). Analysis of nuclear RNA. In RNA Methodologies: A Laboratory Guide for Isolation and Characterization, pp. 236–261. Edited by R. E. Farrell, Jr. San Diego: Academic Press.
  11. Ganem, D. & Varmus, H. E. ( 1987; ). The molecular biology of the hepatitis B viruses. Annual Review of Biochemistry 56, 651-693.[CrossRef]
    [Google Scholar]
  12. Guo, W. , Chen, M. , Yen, T. S. B. & Ou, J.-H. ( 1993; ). Hepatocyte-specific expression of the hepatitis B virus core promoter depends on both positive and negative regulation. Molecular and Cellular Biology 13, 443-448.
    [Google Scholar]
  13. Hennigan, A. N. & Jacobson, A. ( 1996; ). Functional mapping of the translation-dependent instability element of yeast MATα1 mRNA. Molecular and Cellular Biology 16, 3833-3843 .
    [Google Scholar]
  14. Herrick, D. J. & Ross, J. ( 1994; ). The half-life of c-myc mRNA in growing and serum-stimulated cells: influence of the coding and 3′-untranslated regions and role of ribosome translocation. Molecular and Cellular Biology 14, 2119-2128 .
    [Google Scholar]
  15. Hiraga, M. , Nishizono, A. , Mifune, K. , Esumi, M. & Shikata, T. ( 1994; ). Analysis of upstream region of hepatitis B virus core gene using in vitro transcription system. Journal of Medical Virology 43, 404-411.[CrossRef]
    [Google Scholar]
  16. Honigwachs, J. , Faktor, O. , Dickstein, R. , Shaul, Y. & Laub, O. ( 1989; ). Liver-specific expression of hepatitis B virus is determined by the combined action of the core gene promoter and the enhancer. Journal of Virology 63, 919-924.
    [Google Scholar]
  17. Huang, J. & Liang, T. J. ( 1993; ). A novel hepatitis B virus (HBV) genetic element with rev responsive element-like properties that is essential for expression of HBV gene products. Molecular and Cellular Biology 13, 7476-7486 .
    [Google Scholar]
  18. Huang, Z.-M. & Yen, T. S. B. ( 1994; ). Hepatitis B virus RNA element that facilitates accumulation of surface gene transcripts in the cytoplasm. Journal of Virology 68, 3193-3199 .
    [Google Scholar]
  19. Huang, Z.-M. & Yen, T. S. B. ( 1995; ). Role of the hepatitis B virus posttranscriptional regulatory element in export of intronless transcripts. Molecular and Cellular Biology 15, 3864-3869 .
    [Google Scholar]
  20. Huang, Z.-M. , Zang, W.-Q. & Yen, T. S. B. ( 1996; ). Cellular proteins that bind to the hepatitis B virus posttranscriptional regulatory element. Virology 217, 573-581.[CrossRef]
    [Google Scholar]
  21. Kaneko, S. & Miller, R. ( 1988; ). X-region-specific transcripts in mammalian hepatitis B virus-infected liver. Journal of Virology 62, 3979-3984 .
    [Google Scholar]
  22. Kim, Y. H. , Kang, S. K. & Lee, Y. I. ( 1993; ). Functional analysis of hepatitis B virus transactivator X: implication of the leucine zipper like region and C-terminal 7 conserved amino acids in functional regions. Biochemical and Biophysical Research Communications 197, 894-903.[CrossRef]
    [Google Scholar]
  23. Lo, W.-Y. & Ting, L.-P. ( 1994; ). Repression of enhancerII activity by a negative regulatory element in the hepatitis B virus genome. Journal of Virology 68, 1758-1764 .
    [Google Scholar]
  24. Löser, P. , Sandig, V. , Kirrilova, I. & Strauss, M. ( 1996; ). Evaluation of HBV promoters for use in hepatic gene therapy. Biological Chemistry Hoppe-Seyler 377, 187-193.[CrossRef]
    [Google Scholar]
  25. Miller, R. H. ( 1990; ). Organization of the X gene region of the hepatitis B virus genome. Gastroenterologia Japonica 25, 1–5 (Suppl. 2).
    [Google Scholar]
  26. Miller, R. H. & Robinson, W. S. ( 1986; ). Common evolutionary origin of hepatitis B virus and retroviruses. Proceedings of the National Academy of Sciences, USA 83, 2531-2535 .[CrossRef]
    [Google Scholar]
  27. Murakami, S. , Cheong, J. & Kaneko, S. ( 1994; ). Human hepatitis virus X gene encodes a regulatory domain that represses transactivation of X protein. Journal of Biological Chemistry 269, 15118-15123 .
    [Google Scholar]
  28. Nellen, W. & Lichtenstein, C. ( 1993; ). What makes an mRNA anti-sense- itive? Trends in Biochemical Sciences 18, 419-423.[CrossRef]
    [Google Scholar]
  29. Parker, R. & Jacobson, A. ( 1990; ). Translation and a 42-nucleotide segment within the coding region of the mRNA encoded by the MATα1 gene are involved in promoting rapid mRNA decay in yeast. Proceedings of the National Academy of Sciences, USA 87, 2780-2784 .[CrossRef]
    [Google Scholar]
  30. Renner, M. , Haniel, A. , Bürgelt, E. , Hofschneider, P. H. & Koch, W. ( 1995; ). Transactivating function and expression of the X gene of hepatitis B virus. Journal of Hepatology 23, 53-65.[CrossRef]
    [Google Scholar]
  31. Ross, J. ( 1996; ). Control of messenger RNA stability in higher eukaryotes. Trends in Genetics 12, 171-176.[CrossRef]
    [Google Scholar]
  32. Sachs, A. B. ( 1993; ). Messenger RNA degradation in eukaryotes. Cell 74, 413-421.[CrossRef]
    [Google Scholar]
  33. Saiga, A. , Orita, S. , Minoura-Tada, N. , Maeda, M. , Aono, Y. , Asakawa, M. , Nakahara, K. , Kubota, R. , Osame, M. & Igarashi, H. ( 1997; ). Cis-acting inhibitory elements within the pol-env region of human T-cell leukaemia virus type 1 possibly involved in viral persistence. Journal of Virology 71, 4485-4495 .
    [Google Scholar]
  34. Schiavi, S. C. , Wellington, C. L. , Shyu, A.-B. , Chen, C.-Y. A. , Greenberg, M. E. & Belasco, J. B. ( 1994; ). Multiple elements in the c-fos protein coding region facilitate mRNA deadenylation and decay by a mechanism coupled to translation. Journal of Biological Chemistry 269, 3441-3448 .
    [Google Scholar]
  35. Shimoda, A. , Sugata, F. , Chen, H. S. , Miller, R. H. & Purcell, R. H. ( 1998; ). Evidence for a bidirectional promoter complex within the X gene of woodchuck hepatitis virus. Virus Research 56, 25-39.[CrossRef]
    [Google Scholar]
  36. Sokolowski, M. , Tan, W. , Jellne, M. & Schwartz, S. ( 1998; ). mRNA instability elements in the human papillomavirus type 16 L2 coding region. Journal of Virology 72, 1504-1515 .
    [Google Scholar]
  37. Standring, D. N. , Rall, L. B. , Laub, O. & Rutter, W. J. ( 1983; ). Hepatitis B virus encodes an RNA polymerase III transcript. Molecular and Cellular Biology 3, 1774-1782.
    [Google Scholar]
  38. Velhagen, I. , Hilger, C. , Lamberts, C. , Zentgraf, H. & Schröder, C. H. ( 1995; ). An antisense promoter within the hepatitis B virus X gene. Intervirology 38, 127-133.
    [Google Scholar]
  39. Wellington, C. L. , Greenberg, M. E. & Belasco, J. G. ( 1993; ). The destabilizing elements in the coding region of c-fos mRNA are recognized as RNA. Molecular and Cellular Biology 13, 5034-5042 .
    [Google Scholar]
  40. Wisdom, R. & Lee, W. ( 1991; ). The protein-coding region on c-myc mRNA contains a sequence that specifies rapid mRNA turnover and induction by protein synthesis inhibitors. Genes and Development 5, 232-243.[CrossRef]
    [Google Scholar]
  41. Yannish-Perron, C. , Vieira, J. & Messing, J. ( 1985; ). Improved M13 phage cloning vectors and host strains: nucleotide sequence of the M13mp8 and pUC19 vectors. Gene 33, 103-119.[CrossRef]
    [Google Scholar]
  42. Yee, J. K. ( 1989; ). A liver-specific enhancer in the core promoter region of human hepatitis B virus. Science 246, 658-661.[CrossRef]
    [Google Scholar]
  43. Yen, T. J. , Machlin, P. S. & Cleveland, D. W. ( 1988; ). Autoregulated instability of β-tubulin mRNAs by recognition of the nascent amino terminus of β-tubulin. Nature 334, 580-585.[CrossRef]
    [Google Scholar]
  44. Young, R. L. & Korsmeyer, S. J. ( 1993; ). A negative regulatory element in the bcl-2 5′-untranslated region inhibits expression from an upstream promoter. Molecular and Cellular Biology 13, 3686-3697 .
    [Google Scholar]
  45. Yuh, C.-H. & Ting, L.-P. ( 1990; ). The genome of hepatitis B virus contains a second enhancer: cooperation of two elements within this enhancer is required for its function. Journal of Virology 64, 4281-4287 .
    [Google Scholar]
  46. Yuh, C.-H. , Chang, Y.-L. & Ting, L.-P. ( 1992; ). Transcriptional regulation of precore and pregenomic RNAs of hepatitis B virus. Journal of Virology 66, 4073-4084 .
    [Google Scholar]
  47. Zelent, A. Z. , Sells, M. A. , Price, P. M. , Mohamad, A. , Acs, G. & Christman, J. K. ( 1987; ). Murine cells carrying integrated tandem genomes of hepatitis B virus DNA transcribe RNAs from endogenous promoters on both viral strands and express middle and major viral envelope proteins. Journal of Virology 61, 1108-1115 .
    [Google Scholar]
  48. Zhang, P. & McLachlan, A. ( 1994; ). Differentiation-specific transcriptional regulation of the hepatitis B virus nucleocapsid gene in human hepatoma cell lines. Virology 202, 430-440.[CrossRef]
    [Google Scholar]
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