1887

Abstract

The bidirectional activity of the precore/core promoter of hepatitis B virus (HBV) has been demonstrated in cultured cell lines. However, HBV antisense transcripts (asRNAs) have not been demonstrated . In the present study using liver tissue from patients with chronic hepatitis, an anchored 5′RACE mapping the 5′ ends at position 1680/1681, 1655 or 1609/1602 was carried out. In limited cases, RLM-3′RACE detected asRNAs to terminate at four or five consecutive dT residues in the 0·7 kb downstream region. PCR of oligo(dT)-primed cDNA did not amplify a typical polyadenylated asRNA. RT-PCR using various primers did not detect any spliced forms. Competitive RT-PCR estimated the copy numbers of the asRNAs to be 0·05–0·4 % of total sense RNAs. All sequenced asRNAs had ORF6 but, in one patient, the asRNA initiating at position 1680/1681 had additional initiation and termination codons in front of ORF6. Therefore, asRNAs are transcribed by RNA polymerase III at a low level, encompass a dispensable ORF6 gene and might be retained in the nucleus. The endogenous asRNAs complementary to the common ends of all sense RNAs suggest antisense-mediated self-regulation of hepadnavirus.

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2003-07-01
2024-04-19
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References

  1. Ahlquist P. 2002; RNA-dependent RNA polymerases, viruses, and RNA silencing. Science 296:1270–1273
    [Google Scholar]
  2. Chen H. S., Kaneko S., Girones R. 7 other authors 1993; The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks. J Virol 67:1218–1226
    [Google Scholar]
  3. Galibert F., Mandart E., Fitoussi F., Tiollais P., Charnay P. 1979; Nucleotide sequence of the hepatitis B virus genome (subtype ayw ) cloned in E. coli . Nature 281:646–650
    [Google Scholar]
  4. Geiduschek E. P., Tocchini-Valentini G. P. 1988; Transcription by RNA polymerase III. Annu Rev Biochem 57:873–914
    [Google Scholar]
  5. Gunnery S., Mathews M. B. 1995; Functional mRNA can be generated by RNA polymerase III. Mol Cell Biol 15:3597–3607
    [Google Scholar]
  6. Gunther S., Li B. C., Miska S., Kruger D. H., Meisel H., Will H. 1995; A novel method for efficient amplification of whole hepatitis B virus genomes permits rapid functional analysis and reveals deletion mutants in immunosuppressed patients. J Virol 69:5437–5444
    [Google Scholar]
  7. Kumar M., Carmichael G. G. 1998; Antisense RNA: function and fate of duplex RNA in cells of higher eukaryotes. Microbiol Mol Biol Rev 62:1415–1434
    [Google Scholar]
  8. Liu X., Gorovsky M. A. 1993; Mapping the 5′ and 3′ ends of Tetrahymena thermophila mRNAs using RNA ligase-mediated amplification of cDNA ends (RLM-RACE). Nucleic Acids Res 21:4954–4960
    [Google Scholar]
  9. Lo W. Y., Ting L. P. 1994; Repression of enhancer II activity by a negative regulatory element in the hepatitis B virus genome. J Virol 68:1758–1764
    [Google Scholar]
  10. Moriyama K. 1997; Reduced antigen production by hepatitis B virus harbouring nucleotide deletions in the overlapping X gene and precore-core promoter. J Gen Virol 78:1479–1486
    [Google Scholar]
  11. 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 Res 56:25–39
    [Google Scholar]
  12. Standring D. N., Rall L. B., Laub O., Rutter W. J. 1983; Hepatitis B virus encodes an RNA polymerase III transcript. Mol Cell Biol 3:1774–1782
    [Google Scholar]
  13. Velhagen I., Hilger C., Lamberts C., Zentgraf H., Schroder C. H. 1995; An antisense promoter within the hepatitis B virus X gene. Intervirology 38:127–133
    [Google Scholar]
  14. Willis I. M. 1993; RNA polymerase III. Genes, factors and transcriptional specificity. Eur J Biochem 212:1–11
    [Google Scholar]
  15. Yuh C. H., Chang Y. L., Ting L. P. 1992; Transcriptional regulation of precore and pregenomic RNAs of hepatitis B virus. J Virol 66:4073–4084
    [Google Scholar]
  16. 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. J Virol 61:1108–1115
    [Google Scholar]
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