Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1990 Feb;9(2):497–504. doi: 10.1002/j.1460-2075.1990.tb08136.x

The hepatitis B virus X-gene product trans-activates both RNA polymerase II and III promoters.

B Aufiero 1, R J Schneider 1
PMCID: PMC551692  PMID: 2303039

Abstract

The transcriptional regulatory activity of the human hepatitis B virus (HBV) X-gene product was investigated. We demonstrate a new property for the HBV X-gene, the strong transcriptional trans-activation of promoters for class III genes. The stimulation of RNA polymerase III (pol III) as well as pol II promoters is shown in cells transiently transfected with the X-gene, and after its stable integration into hepatocytes. We demonstrate that X-gene containing cells stimulate the frequency of pol III transcription initiation by 20- to 40-fold, and accelerate the rate of formation of stable pol III initiation complexes in a manner indistinguishable from that of adenovirus E1a protein. Since the transcription factor TFIIIC has been shown to be limiting in the formation of stable pol III initiation complexes, template commitment experiments were performed which titrate the level of this factor in extracts. We show that X-protein containing extracts are far more efficient in forming stable pol III preinitiation complexes that cannot be competed away upon addition of a second template, indicating that TFIIIC is very probably a target of the X-protein. Thus, the HBV X-protein is apparently a member of a family of trans-activators capable of stimulating both pol II and III promoters, which includes the adenovirus E1a-protein and SV40 t antigen.

Full text

PDF
502

Images in this article

498Fig. 1.
on p.498
498Fig. 2.
on p.498
499Fig. 3.
on p.499
499Fig. 4.
on p.499
500Fig. 5.
on p.500
500Fig. 6.
on p.500
501Fig. 7.
on p.501
501Fig. 8.
on p.501

Selected References

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

  1. Berger S. L., Folk W. R. Differential activation of RNA polymerase III-transcribed genes by the polyomavirus enhancer and the adenovirus E1A gene products. Nucleic Acids Res. 1985 Feb 25;13(4):1413–1428. doi: 10.1093/nar/13.4.1413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Carey M. F., Singh K., Botchan M., Cozzarelli N. R. Induction of specific transcription by RNA polymerase III in transformed cells. Mol Cell Biol. 1986 Sep;6(9):3068–3076. doi: 10.1128/mcb.6.9.3068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chisaka O., Araki K., Ochiya T., Tsurimoto T., Hiranyawasitte-Attatippaholkun W., Yanaihara N., Matsubara K. Purification of hepatitis B virus gene X product synthesized in Escherichia coli and its detection in a human hepatoblastoma cell line producing hepatitis B virus. Gene. 1987;60(2-3):183–189. doi: 10.1016/0378-1119(87)90226-5. [DOI] [PubMed] [Google Scholar]
  5. Colbère-Garapin F., Horodniceanu F., Kourilsky P., Garapin A. C. A new dominant hybrid selective marker for higher eukaryotic cells. J Mol Biol. 1981 Jul 25;150(1):1–14. doi: 10.1016/0022-2836(81)90321-1. [DOI] [PubMed] [Google Scholar]
  6. Dean N., Berk A. J. Ordering promoter binding of class III transcription factors TFIIIC1 and TFIIIC2. Mol Cell Biol. 1988 Aug;8(8):3017–3025. doi: 10.1128/mcb.8.8.3017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dejean A., Bougueleret L., Grzeschik K. H., Tiollais P. Hepatitis B virus DNA integration in a sequence homologous to v-erb-A and steroid receptor genes in a hepatocellular carcinoma. Nature. 1986 Jul 3;322(6074):70–72. doi: 10.1038/322070a0. [DOI] [PubMed] [Google Scholar]
  8. Edwards D. R., Parfett C. L., Denhardt D. T. Transcriptional regulation of two serum-induced RNAs in mouse fibroblasts: equivalence of one species to B2 repetitive elements. Mol Cell Biol. 1985 Nov;5(11):3280–3288. doi: 10.1128/mcb.5.11.3280. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Elfassi E., Haseltine W. A., Dienstag J. L. Detection of hepatitis B virus X product using an open reading frame Escherichia coli expression vector. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2219–2222. doi: 10.1073/pnas.83.7.2219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  12. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Fu X. X., Su C. Y., Lee Y., Hintz R., Biempica L., Snyder R., Rogler C. E. Insulinlike growth factor II expression and oval cell proliferation associated with hepatocarcinogenesis in woodchuck hepatitis virus carriers. J Virol. 1988 Sep;62(9):3422–3430. doi: 10.1128/jvi.62.9.3422-3430.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fuhrman S. A., Engelke D. R., Geiduschek E. P. HeLa cell RNA polymerase III transcription factors. Functional characterization of a fraction identified by its activity in a second template rescue assay. J Biol Chem. 1984 Feb 10;259(3):1934–1943. [PubMed] [Google Scholar]
  15. Ganem D., Varmus H. E. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. doi: 10.1146/annurev.bi.56.070187.003251. [DOI] [PubMed] [Google Scholar]
  16. Gaynor R. B., Feldman L. T., Berk A. J. Transcription of class III genes activated by viral immediate early proteins. Science. 1985 Oct 25;230(4724):447–450. doi: 10.1126/science.2996135. [DOI] [PubMed] [Google Scholar]
  17. Geiduschek E. P., Tocchini-Valentini G. P. Transcription by RNA polymerase III. Annu Rev Biochem. 1988;57:873–914. doi: 10.1146/annurev.bi.57.070188.004301. [DOI] [PubMed] [Google Scholar]
  18. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  20. Greenberg M. E., Ziff E. B. Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene. Nature. 1984 Oct 4;311(5985):433–438. doi: 10.1038/311433a0. [DOI] [PubMed] [Google Scholar]
  21. Hoeffler W. K., Kovelman R., Roeder R. G. Activation of transcription factor IIIC by the adenovirus E1A protein. Cell. 1988 Jun 17;53(6):907–920. doi: 10.1016/s0092-8674(88)90409-6. [DOI] [PubMed] [Google Scholar]
  22. Hoeffler W. K., Roeder R. G. Enhancement of RNA polymerase III transcription by the E1A gene product of adenovirus. Cell. 1985 Jul;41(3):955–963. doi: 10.1016/s0092-8674(85)80076-3. [DOI] [PubMed] [Google Scholar]
  23. Hsu T., Möröy T., Etiemble J., Louise A., Trépo C., Tiollais P., Buendia M. A. Activation of c-myc by woodchuck hepatitis virus insertion in hepatocellular carcinoma. Cell. 1988 Nov 18;55(4):627–635. doi: 10.1016/0092-8674(88)90221-8. [DOI] [PubMed] [Google Scholar]
  24. Kaneko S., Miller R. H. X-region-specific transcript in mammalian hepatitis B virus-infected liver. J Virol. 1988 Nov;62(11):3979–3984. doi: 10.1128/jvi.62.11.3979-3984.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kay A., Mandart E., Trepo C., Galibert F. The HBV HBX gene expressed in E. coli is recognised by sera from hepatitis patients. EMBO J. 1985 May;4(5):1287–1292. doi: 10.1002/j.1460-2075.1985.tb03774.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lassar A. B., Martin P. L., Roeder R. G. Transcription of class III genes: formation of preinitiation complexes. Science. 1983 Nov 18;222(4625):740–748. doi: 10.1126/science.6356356. [DOI] [PubMed] [Google Scholar]
  27. Lo S. J., Chien M. L., Lee Y. H. Characteristics of the X gene of hepatitis B virus. Virology. 1988 Nov;167(1):289–292. doi: 10.1016/0042-6822(88)90081-5. [DOI] [PubMed] [Google Scholar]
  28. Loeken M. R., Khoury G., Brady J. Stimulation of the adenovirus E2 promoter by simian virus 40 T antigen or E1A occurs by different mechanisms. Mol Cell Biol. 1986 Jun;6(6):2020–2026. doi: 10.1128/mcb.6.6.2020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Meyers M. L., Trepo L. V., Nath N., Sninsky J. J. Hepatitis B virus polypeptide X: expression in Escherichia coli and identification of specific antibodies in sera from hepatitis B virus-infected humans. J Virol. 1986 Jan;57(1):101–109. doi: 10.1128/jvi.57.1.101-109.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Miller R. H., Robinson W. S. Common evolutionary origin of hepatitis B virus and retroviruses. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2531–2535. doi: 10.1073/pnas.83.8.2531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Miyaki M., Sato C., Gotanda T., Matsui T., Mishiro S., Imai M., Mayumi M. Integration of region X of hepatitis B virus genome in human primary hepatocellular carcinomas propagated in nude mice. J Gen Virol. 1986 Jul;67(Pt 7):1449–1454. doi: 10.1099/0022-1317-67-7-1449. [DOI] [PubMed] [Google Scholar]
  32. Moriarty A. M., Alexander H., Lerner R. A., Thornton G. B. Antibodies to peptides detect new hepatitis B antigen: serological correlation with hepatocellular carcinoma. Science. 1985 Jan 25;227(4685):429–433. doi: 10.1126/science.2981434. [DOI] [PubMed] [Google Scholar]
  33. Nagaya T., Nakamura T., Tokino T., Tsurimoto T., Imai M., Mayumi T., Kamino K., Yamamura K., Matsubara K. The mode of hepatitis B virus DNA integration in chromosomes of human hepatocellular carcinoma. Genes Dev. 1987 Oct;1(8):773–782. doi: 10.1101/gad.1.8.773. [DOI] [PubMed] [Google Scholar]
  34. Ochiya T., Fujiyama A., Fukushige S., Hatada I., Matsubara K. Molecular cloning of an oncogene from a human hepatocellular carcinoma. Proc Natl Acad Sci U S A. 1986 Jul;83(14):4993–4997. doi: 10.1073/pnas.83.14.4993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Persing D. H., Varmus H. E., Ganem D. Antibodies to pre-S and X determinants arise during natural infection with ground squirrel hepatitis virus. J Virol. 1986 Oct;60(1):177–184. doi: 10.1128/jvi.60.1.177-184.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Popper H., Shafritz D. A., Hoofnagle J. H. Relation of the hepatitis B virus carrier state to hepatocellular carcinoma. Hepatology. 1987 Jul-Aug;7(4):764–772. doi: 10.1002/hep.1840070425. [DOI] [PubMed] [Google Scholar]
  37. Schneider R. J., Shenk T. Impact of virus infection on host cell protein synthesis. Annu Rev Biochem. 1987;56:317–332. doi: 10.1146/annurev.bi.56.070187.001533. [DOI] [PubMed] [Google Scholar]
  38. Shaul Y., Ziemer M., Garcia P. D., Crawford R., Hsu H., Valenzuela P., Rutter W. J. Cloning and analysis of integrated hepatitis virus sequences from a human hepatoma cell line. J Virol. 1984 Sep;51(3):776–787. doi: 10.1128/jvi.51.3.776-787.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Siddiqui A., Jameel S., Mapoles J. Expression of the hepatitis B virus X gene in mammalian cells. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2513–2517. doi: 10.1073/pnas.84.8.2513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Singh K., Carey M., Saragosti S., Botchan M. Expression of enhanced levels of small RNA polymerase III transcripts encoded by the B2 repeats in simian virus 40-transformed mouse cells. Nature. 1985 Apr 11;314(6011):553–556. doi: 10.1038/314553a0. [DOI] [PubMed] [Google Scholar]
  41. Spandau D. F., Lee C. H. trans-activation of viral enhancers by the hepatitis B virus X protein. J Virol. 1988 Feb;62(2):427–434. doi: 10.1128/jvi.62.2.427-434.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Standring D. N., Rall L. B., Laub O., Rutter W. J. Hepatitis B virus encodes an RNA polymerase III transcript. Mol Cell Biol. 1983 Oct;3(10):1774–1782. doi: 10.1128/mcb.3.10.1774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. Twu J. S., Schloemer R. H. Transcriptional trans-activating function of hepatitis B virus. J Virol. 1987 Nov;61(11):3448–3453. doi: 10.1128/jvi.61.11.3448-3453.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Velcich A., Ziff E. Adenovirus E1a proteins repress transcription from the SV40 early promoter. Cell. 1985 Mar;40(3):705–716. doi: 10.1016/0092-8674(85)90219-3. [DOI] [PubMed] [Google Scholar]
  46. Weinmann R., Jaehning J. A., Raskas H. J., Roeder R. G. Viral RNA synthesis and levels of DNA-dependent RNA polymerases during replication of adenovirus 2. J Virol. 1975 Jan;17(1):114–126. doi: 10.1128/jvi.17.1.114-126.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wigler M., Pellicer A., Silverstein S., Axel R. Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978 Jul;14(3):725–731. doi: 10.1016/0092-8674(78)90254-4. [DOI] [PubMed] [Google Scholar]
  48. Wollersheim M., Debelka U., Hofschneider P. H. A transactivating function encoded in the hepatitis B virus X gene is conserved in the integrated state. Oncogene. 1988 Nov;3(5):545–552. [PubMed] [Google Scholar]
  49. Yoshinaga S. K., L'Etoile N. D., Berk A. J. Purification and characterization of transcription factor IIIC2. J Biol Chem. 1989 Jun 25;264(18):10726–10731. [PubMed] [Google Scholar]
  50. Yoshinaga S., Dean N., Han M., Berk A. J. Adenovirus stimulation of transcription by RNA polymerase III: evidence for an E1A-dependent increase in transcription factor IIIC concentration. EMBO J. 1986 Feb;5(2):343–354. doi: 10.1002/j.1460-2075.1986.tb04218.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zahm P., Hofschneider P. H., Koshy R. The HBV X-ORF encodes a transactivator: a potential factor in viral hepatocarcinogenesis. Oncogene. 1988 Aug;3(2):169–177. [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES