1887

Abstract

We have previously cloned a mutant hepatitis B virus (HBV) genome which had one thymidine addition in the pre-C region resulting in a frameshift mutation in the pre-C region and fusion of the X and C genes. We constructed plasmids containing serially deleted and/or back-mutated (authentic) pre-C regions to study the effect of the frameshift mutation. COS cells transfected with plasmids containing the frameshifted pre-C region produced a 21K C protein (P21c) but not a 22K partially processed pre-C protein (P22). On the other hand, COS cells transfected with plasmids containing the back-mutated pre-C region produced P22. This result was also observed in HepG2-K8 cells producing the mutant HBV particles. Therefore, the pre-C region of HBV is likely to be non-essential for virus replication. COS cells transfected with the plasmid containing a fused X-C open reading frame (ORF) produced a 40K X-C fusion protein. This X-C fusion protein exerted transcriptional trans-activation. These results suggest that the mutant HBV has a C gene with a defective pre-C region and a fused X-C ORF, and hence cannot synthesize 16K HBeAg (P16e).

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-75-4-917
1994-04-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/75/4/JV0750040917.html?itemId=/content/journal/jgv/10.1099/0022-1317-75-4-917&mimeType=html&fmt=ahah

References

  1. Bchini R., Capel F., Dauguet C., Dubanchet S., Petit M.-A. 1990; In vitro infection of human hepatoma (HepG2) cells with hepatitis B virus. Journal of Virology 64:3025–3032
    [Google Scholar]
  2. Brunetto M. R., Stemler M., Schodel F., Will H., Ottoberlli A., Rizzetto M., Bonino F. 1989; Identification of HBV variants which cannot produce precore derived HBeAg and may be responsible for severe hepatitis. Italian Journal of Gastroenterology 21:151–154
    [Google Scholar]
  3. Brunetto M. R., Stemler M., Bonino F., Schodel F., Oliveri F., Rizzetto M., Verme G., Will H. 1990; A new hepatitis B virus-strain in patients with severe anti-HBe positive chronic hepatitis. Journal of Hepatology 10:258–261
    [Google Scholar]
  4. Bruss V., Gerlich W. H. 1988; Formation of transmembraneous hepatitis B e-antigen by cotranslational in vitro processing of the viral precore protein. Virology 163:268–275
    [Google Scholar]
  5. Carman W. F., Jacyna M. R., Hadziyannis S., Karayiannis P., Mcgarvey M. J., Markris A., Thomas H. C. 1989; The mutation preventing formation of hepatitis B e antigen in patients with chronic hepatitis B infection. Lancet ii:585–591
    [Google Scholar]
  6. Chang C., Enders G., Sprengel R., Peters N., Varmus H. E., Ganem D. 1987; Expression of the precore region of an avian hepatitis B virus is not required for viral replication. Journal of Virology 61:3322–3325
    [Google Scholar]
  7. Chen C., Okayama H. 1987; High-efficiency transformation of mammalian cells by plasmid DNA. Molecular and Cellular Biology 7:2745–2752
    [Google Scholar]
  8. Choi M Y., Kim Y. S., Rho H. M. 1984; Cloning and genetic mapping of the hepatitis B viral genome. Journal of the Korean Society of Virology 14:49–55
    [Google Scholar]
  9. Choi S. G., Lee w. S., Kim S. K., Rho H. M. 1991; Expression of hepatitis B viral core antigen gene in Escherichia coli. Korean Journal of Microbiology 29:80–84
    [Google Scholar]
  10. Farza H., Hachouel M., Scotto J., Tiollais P., Babinet C., Pourcel C. 1988; Replication and gene expression of hepatitis B virus in a transgenic mouse that contains the complete viral genome. Journal of Virology 62:4144–4152
    [Google Scholar]
  11. Feitelson M. A. 1986; Products of the X gene in hepatitis B and related viruses. Hepatology 6:191–198
    [Google Scholar]
  12. Feitelson M. A., Miller R. H. 1988; X gene-related sequences in the core gene of duck and heron hepatitis B viruses. Proceedings of the National Academy of Sciences, U.S.A 85:6162–6166
    [Google Scholar]
  13. Fiordalisi G., Cariani E., Mantero G., Zanetti A., Tanzi E., Chiaramonte M., Primi D. 1990; High genomic variability in the pre-C region of hepatitis B virus in anti-HBe, HBV DNApositive chronic hepatitis. Journal of Medical Virology 31:297–300
    [Google Scholar]
  14. Fitzgerald M., Shenk T. 1981; The sequence 5′-AAUAAA-3′ forms part of the recognition site for polyadenylation of late SV40 mRNAs. Cell 24:251–260
    [Google Scholar]
  15. Ganem D., Varmus H. E. 1987; The molecular biology of the hepatitis B virus. Annual Review of Biochemistry 56:651–693
    [Google Scholar]
  16. Garcia P. D., Ou J.-H., Rutter W. J., Walter P. 1988; Targeting of the hepatitis B virus precore protein to the endoplasmic reticulum membrane: after signal peptide cleavage translocation can be aborted and the product released into the cytoplasm. Journal of Cell Biology 106:1093–1104
    [Google Scholar]
  17. Gorman C. M., Moffat L. F., Howard B. H. 1982; Recombinant genomes express chloramphenicol acetyltransferase in mammalian cells. Molecular and Cellular Biology 2:1044–1051
    [Google Scholar]
  18. Guo W., Wang J., Tam G., Yen T. S. B., Ou J.-H. 1991; Leaky transcription termination produces larger and smaller than genome size hepatitis B virus X gene transcripts. Virology 181:630–636
    [Google Scholar]
  19. Hoofnagle J., Dusheiko G. M., Seel L. B., Jones L. B., Waggoner J. G., Bales Z. B. 1981; Seroconversion from hepatitis B e antigen to antibody in chronic type B hepatitis. Annals of Internal Medicine 94:744–748
    [Google Scholar]
  20. Junker M., Galle P., Schaller H. 1987; Expression and replication of the hepatitis B virus genome under foreign promoter control. Nucleic Acids Research 15:10117–10132
    [Google Scholar]
  21. Kammann M., Laufs J., Schell J., Gronanborn B. 1989; Rapid insertional mutagenesis of DNA by polymerase chain reaction (PCR). Nucleic Acids Research 17:5404
    [Google Scholar]
  22. Kay A., Mandart E., Trepo C., Galibert F. 1985; The HBV HBx gene expressed in E. coli is recognised by sera from hepatitis patients. EMBO Journal 4:1287–1292
    [Google Scholar]
  23. Kim S. K., Rho H. M. 1992; Purification and characterization of the hepatitis B virus core antigen gene product expressed in Escherichia coli. Korean Biochemical Journal 25:733–738
    [Google Scholar]
  24. Kim S. H., Hong S. P., Kim S. K., Lee W. S., Rho H. M. 1992a; Replication of a mutant hepatitis B virus with a fused X-C reading frame in hepatoma cells. Journal of General Virology 73:2421–2424
    [Google Scholar]
  25. Kim S. K., Shim J. W., Rho H. M. 1992b; Effect of the mutation in the carboxyl-terminal processing site of the hepatitis B virus core antigen on the HBeAg secretion. Journal of the Korean Society of Virology 22:97–104
    [Google Scholar]
  26. Kozak M. 1986; Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell 44:283–292
    [Google Scholar]
  27. Levrero M., Balsand C., Natoli G., Avantaggiati M. L., Elfassi E. 1990; Hepatitis B virus X protein transactivates the long terminal repeats of human immunodeficiency virus types 1 and 2. Journal of Virology 64:3082–3086
    [Google Scholar]
  28. Li J. S., Tong S., Vitvitski L., Zoulim F., Trépo C. 1990; Rapid detection and further characterization of infection with hepatitis B virus variants containing a stop codon in the distal pre-C region. Journal of General Virology 71:1993–1998
    [Google Scholar]
  29. Lo S. J., You L.-R., Lee Y.-H. W. 1990; The hepatitis B virus X-C fusion protein is unlikely to be produced by the mechanism of ribosomal frameshifting. Virology 178:584–587
    [Google Scholar]
  30. Mandart E., Kay A., Galibert F. 1984; Nucleotide sequence of a cloned duck hepatitis B virus genome: comparison with woodchuck and human hepatitis B virus sequences. Journal of Virology 49:782–792
    [Google Scholar]
  31. Meyers M. L., Trepo L. V., Nath N., Sninsky J. J. 1986; Hepatitis B virus polypeptide X: expression in Escherichia coli and identification of specific antibodies in sera from hepatitis B virus-infected humans. Journal of Virology 57:101–109
    [Google Scholar]
  32. Montell C., Fisher E. F., Caruthers M. H., Berk A. J. 1983; Inhibition of RNA cleavage but not polyadenylation by a point mutation in mRNA 3′ consensus sequence AAUAAA. Nature; London: 305600–605
    [Google Scholar]
  33. Moriarty A, Alexander H., Lerner R. A., Thornton G. B. 1985; Antibodies to peptides detect new hepatitis B antigen: serological correlation with hepatocellular carcinoma. Science 227:429–432
    [Google Scholar]
  34. Okamoto H., Yotsumoto S., Akahane Y., Yamanaka T., Miyazaki Y., Sugai Y., Tsuda F., Tanaka T., Miyakawa Y., Mayumi M. 1990; Hepatitis B viruses with precore region defects prevail in persistently infected hosts along with seroconversion to the antibody against e antigen. Journal of Virology 64:1298–1303
    [Google Scholar]
  35. Ou J. H., Laub O., Rutter W. J. 1986; Hepatitis B virus gene function: precore region targets the core antigen to cellular membranes and causes the secretion of the e antigen. Proceedings of the National Academy of Sciences, U.S.A 83:1578–1582
    [Google Scholar]
  36. Ou J. H., Standring D. N., Masiarz F. R., Rutter W. J. 1988; A signal peptide encoded within the precore region of the hepatitis B virus directs the secretion of a heterogeneous population of e antigens in Xenopus oocytes. Proceedings of the National Academy of Sciences, U.S.A 85:8405–8409
    [Google Scholar]
  37. Realdi G., Alberh A., Rugge F., Bortoloth F., Rigoh A. M., Tremolada F., Ruol A. 1980; Seroconversion from hepatitis B e antigen to anti-HBe in chronic hepatitis B virus infection. Gastroenterology 79:195–199
    [Google Scholar]
  38. Rho H. M., Kim K. T., Hyun S. W., Kim Y. S. 1989; The nucleotide sequence and reading frames of a mutant hepatitis B virus subtype adr. Nucleic Acids Research 17:2124
    [Google Scholar]
  39. Riamondo G., Schneider R., Stemler M., Smedile V., Rodino G., Will H. 1990; A new hepatitis B virus variant in a chronic carrier with multiple episodes of viral reactivation and acute hepatitis. Virology 179:64–68
    [Google Scholar]
  40. Ritter S. E., Whitten T. M., Quets A. T., Schlommer R. H. 1991; An internal domain of the hepatitis B virus X is necessary for transacting activity. Virology 182:841–845
    [Google Scholar]
  41. Roossinck M. J., Jameel S., Louskin S. H., Siddiqui A. 1986; Expression of hepatitis B core region in mammalian cells. Molecular and Cellular Biology 6:1393–1400
    [Google Scholar]
  42. Saiki R. K., Gelfand D. H., Stoffel S., Scharf S. J., Higuchi R., Horn G. T., Mullis K. B., Erlich H. A. 1988; Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science 239:487–491
    [Google Scholar]
  43. Salfeld J., Pfaff E., Noah M., Schaller H. 1989; Antigenic determination and functional domains in core antigen and e antigen from hepatitis B virus. Journal of Virology 63:798–808
    [Google Scholar]
  44. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual, 2nd edn. New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  45. Schlicht H. J., Salfeld J., Schaller H. 1987; 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. Journal of Virology 61:3701–3709
    [Google Scholar]
  46. Spandau D. F., Lee C.-H. 1988; Trans-activation of viral enhancers by the hepatitis B virus X protein. Journal of Virology 62:427–434
    [Google Scholar]
  47. Tiollais P., Pourcel C., Dejean A. 1985; The hepatitis B virus. Nature; London: 317489–495
    [Google Scholar]
  48. Tong S. P., Li J. S., Vitvitski L., Trepo C. 1990; Active hepatitis B virus replication in the presence of anti-HBe is associated with viral variants containing an inactive pre-C region. Virology 176:596–603
    [Google Scholar]
  49. Tong S. P., Diot C., Gripon P., Li J. S., Vitvitski L., Trepo C., Guguen-Guillouzo C. 1991; In vitro replication competence of a cloned hepatitis B virus variant with a nonsense mutation in the distal pre-C region. Virology 181:733–737
    [Google Scholar]
  50. Twu J.-S., Schloemer R. H. 1987; Transcriptional trans activating function of hepatitis B virus. Journal of Virology 61:3448–3453
    [Google Scholar]
  51. Valenzuela P., Quiroga M., Zaldivar J., Gray P., Rutter W. J. 1980; The nucleotide sequence of the hepatitis B viral genome and the identification of major viral genes. In Animal Virus Genetics pp. 57–70 Fields B. N., Jaenisch R., Fox L. Edited by New York: Academic Press;
    [Google Scholar]
  52. Weimer T., Salfeld J., Will H. 1987; Expression of the hepatitis B viral core gene in vitro and in vivo. Journal of Virology 6:3109–3113
    [Google Scholar]
  53. Won H. J., Yoon J. H., Choi C. Y., Sun H. S., Rho H. M. 1989; Immunoprecipitation of the HBV X-gene product by sera from hepatitis B virus-infected patient. Journal of the Korean Society of Virology 19:151–157
    [Google Scholar]
  54. Zahm P., Fiofschneider P. H., Koshy R. 1988; The HBV-X ORF encodes a transactivator: a potential factor in viral hepatocarcinogenesis. Oncogene 3:169–177
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-75-4-917
Loading
/content/journal/jgv/10.1099/0022-1317-75-4-917
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error