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. 1997 Aug;71(8):6194–6199. doi: 10.1128/jvi.71.8.6194-6199.1997

Interaction of the UV-damaged DNA-binding protein with hepatitis B virus X protein is conserved among mammalian hepadnaviruses and restricted to transactivation-proficient X-insertion mutants.

D Sitterlin 1, T H Lee 1, S Prigent 1, P Tiollais 1, J S Butel 1, C Transy 1
PMCID: PMC191882  PMID: 9223516

Abstract

We carried out a comparative analysis of several proposed host protein partners of the human hepatitis B virus X protein (HBx) using both the GAL4- and the LexA-based yeast two-hybrid system. We showed that the interaction of HBx with the UV-damaged DNA-binding protein (UVDDB) is positive in both yeast systems, detectable in cotransfected human cells, conserved by rodent hepadnavirus X proteins (known to transactivate in human cells), and tightly correlated with the transactivation proficiency of X-insertion mutants. Taken together, our results strongly suggest that UVDDB is involved in X-mediated transactivation.

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Selected References

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  1. Arii M., Takada S., Koike K. Identification of three essential regions of hepatitis B virus X protein for trans-activation function. Oncogene. 1992 Mar;7(3):397–403. [PubMed] [Google Scholar]
  2. Beasley R. P., Hwang L. Y., Lin C. C., Chien C. S. Hepatocellular carcinoma and hepatitis B virus. A prospective study of 22 707 men in Taiwan. Lancet. 1981 Nov 21;2(8256):1129–1133. doi: 10.1016/s0140-6736(81)90585-7. [DOI] [PubMed] [Google Scholar]
  3. Benn J., Schneider R. J. Hepatitis B virus HBx protein activates Ras-GTP complex formation and establishes a Ras, Raf, MAP kinase signaling cascade. Proc Natl Acad Sci U S A. 1994 Oct 25;91(22):10350–10354. doi: 10.1073/pnas.91.22.10350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blum H. E., Zhang Z. S., Galun E., von Weizsäcker F., Garner B., Liang T. J., Wands J. R. Hepatitis B virus X protein is not central to the viral life cycle in vitro. J Virol. 1992 Feb;66(2):1223–1227. doi: 10.1128/jvi.66.2.1223-1227.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Buendia M. A. Hepatitis B viruses and hepatocellular carcinoma. Adv Cancer Res. 1992;59:167–226. doi: 10.1016/s0065-230x(08)60306-1. [DOI] [PubMed] [Google Scholar]
  6. Chen H. S., Kaneko S., Girones R., Anderson R. W., Hornbuckle W. E., Tennant B. C., Cote P. J., Gerin J. L., Purcell R. H., Miller R. H. The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks. J Virol. 1993 Mar;67(3):1218–1226. doi: 10.1128/jvi.67.3.1218-1226.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cheong J. H., Yi M., Lin Y., Murakami S. Human RPB5, a subunit shared by eukaryotic nuclear RNA polymerases, binds human hepatitis B virus X protein and may play a role in X transactivation. EMBO J. 1995 Jan 3;14(1):143–150. doi: 10.1002/j.1460-2075.1995.tb06984.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Colgrove R., Simon G., Ganem D. Transcriptional activation of homologous and heterologous genes by the hepatitis B virus X gene product in cells permissive for viral replication. J Virol. 1989 Sep;63(9):4019–4026. doi: 10.1128/jvi.63.9.4019-4026.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cross J. C., Wen P., Rutter W. J. Transactivation by hepatitis B virus X protein is promiscuous and dependent on mitogen-activated cellular serine/threonine kinases. Proc Natl Acad Sci U S A. 1993 Sep 1;90(17):8078–8082. doi: 10.1073/pnas.90.17.8078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doria M., Klein N., Lucito R., Schneider R. J. The hepatitis B virus HBx protein is a dual specificity cytoplasmic activator of Ras and nuclear activator of transcription factors. EMBO J. 1995 Oct 2;14(19):4747–4757. doi: 10.1002/j.1460-2075.1995.tb00156.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fagan R., Flint K. J., Jones N. Phosphorylation of E2F-1 modulates its interaction with the retinoblastoma gene product and the adenoviral E4 19 kDa protein. Cell. 1994 Sep 9;78(5):799–811. doi: 10.1016/s0092-8674(94)90522-3. [DOI] [PubMed] [Google Scholar]
  12. Feitelson M. A., Zhu M., Duan L. X., London W. T. Hepatitis B x antigen and p53 are associated in vitro and in liver tissues from patients with primary hepatocellular carcinoma. Oncogene. 1993 May;8(5):1109–1117. [PubMed] [Google Scholar]
  13. Fields S., Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989 Jul 20;340(6230):245–246. doi: 10.1038/340245a0. [DOI] [PubMed] [Google Scholar]
  14. Fischer M., Runkel L., Schaller H. HBx protein of hepatitis B virus interacts with the C-terminal portion of a novel human proteasome alpha-subunit. Virus Genes. 1995;10(1):99–102. doi: 10.1007/BF01724303. [DOI] [PubMed] [Google Scholar]
  15. Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. doi: 10.1016/0076-6879(83)01013-7. [DOI] [PubMed] [Google Scholar]
  16. Gyuris J., Golemis E., Chertkov H., Brent R. Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2. Cell. 1993 Nov 19;75(4):791–803. doi: 10.1016/0092-8674(93)90498-f. [DOI] [PubMed] [Google Scholar]
  17. Harper J. W., Adami G. R., Wei N., Keyomarsi K., Elledge S. J. The p21 Cdk-interacting protein Cip1 is a potent inhibitor of G1 cyclin-dependent kinases. Cell. 1993 Nov 19;75(4):805–816. doi: 10.1016/0092-8674(93)90499-g. [DOI] [PubMed] [Google Scholar]
  18. Haviv I., Vaizel D., Shaul Y. pX, the HBV-encoded coactivator, interacts with components of the transcription machinery and stimulates transcription in a TAF-independent manner. EMBO J. 1996 Jul 1;15(13):3413–3420. [PMC free article] [PubMed] [Google Scholar]
  19. Hollenberg S. M., Sternglanz R., Cheng P. F., Weintraub H. Identification of a new family of tissue-specific basic helix-loop-helix proteins with a two-hybrid system. Mol Cell Biol. 1995 Jul;15(7):3813–3822. doi: 10.1128/mcb.15.7.3813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Huang J., Kwong J., Sun E. C., Liang T. J. Proteasome complex as a potential cellular target of hepatitis B virus X protein. J Virol. 1996 Aug;70(8):5582–5591. doi: 10.1128/jvi.70.8.5582-5591.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hwang B. J., Chu G. Purification and characterization of a human protein that binds to damaged DNA. Biochemistry. 1993 Feb 16;32(6):1657–1666. doi: 10.1021/bi00057a033. [DOI] [PubMed] [Google Scholar]
  22. Kazantsev A., Mu D., Nichols A. F., Zhao X., Linn S., Sancar A. Functional complementation of xeroderma pigmentosum complementation group E by replication protein A in an in vitro system. Proc Natl Acad Sci U S A. 1996 May 14;93(10):5014–5018. doi: 10.1073/pnas.93.10.5014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kekulé A. S., Lauer U., Weiss L., Luber B., Hofschneider P. H. Hepatitis B virus transactivator HBx uses a tumour promoter signalling pathway. Nature. 1993 Feb 25;361(6414):742–745. doi: 10.1038/361742a0. [DOI] [PubMed] [Google Scholar]
  24. Kumar V., Jayasuryan N., Kumar R. A truncated mutant (residues 58-140) of the hepatitis B virus X protein retains transactivation function. Proc Natl Acad Sci U S A. 1996 May 28;93(11):5647–5652. doi: 10.1073/pnas.93.11.5647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lee T. H., Elledge S. J., Butel J. S. Hepatitis B virus X protein interacts with a probable cellular DNA repair protein. J Virol. 1995 Feb;69(2):1107–1114. doi: 10.1128/jvi.69.2.1107-1114.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Legrain P., Chapon C. Interaction between PRP11 and SPP91 yeast splicing factors and characterization of a PRP9-PRP11-SPP91 complex. Science. 1993 Oct 1;262(5130):108–110. doi: 10.1126/science.8211114. [DOI] [PubMed] [Google Scholar]
  27. Maguire H. F., Hoeffler J. P., Siddiqui A. HBV X protein alters the DNA binding specificity of CREB and ATF-2 by protein-protein interactions. Science. 1991 May 10;252(5007):842–844. doi: 10.1126/science.1827531. [DOI] [PubMed] [Google Scholar]
  28. Nassal M., Schaller H. Hepatitis B virus replication--an update. J Viral Hepat. 1996 Sep;3(5):217–226. doi: 10.1111/j.1365-2893.1996.tb00047.x. [DOI] [PubMed] [Google Scholar]
  29. Qadri I., Ferrari M. E., Siddiqui A. The hepatitis B virus transactivator protein, HBx, interacts with single-stranded DNA (ssDNA). Biochemical characterizations of the HBx-ssDNA interactions. J Biol Chem. 1996 Jun 28;271(26):15443–15450. doi: 10.1074/jbc.271.26.15443. [DOI] [PubMed] [Google Scholar]
  30. Rossner M. T. Review: hepatitis B virus X-gene product: a promiscuous transcriptional activator. J Med Virol. 1992 Feb;36(2):101–117. doi: 10.1002/jmv.1890360207. [DOI] [PubMed] [Google Scholar]
  31. Runkel L., Fischer M., Schaller H. Two-codon insertion mutations of the HBx define two separate regions necessary for its trans-activation function. Virology. 1993 Dec;197(2):529–536. doi: 10.1006/viro.1993.1626. [DOI] [PubMed] [Google Scholar]
  32. Takao M., Abramic M., Moos M., Jr, Otrin V. R., Wootton J. C., McLenigan M., Levine A. S., Protic M. A 127 kDa component of a UV-damaged DNA-binding complex, which is defective in some xeroderma pigmentosum group E patients, is homologous to a slime mold protein. Nucleic Acids Res. 1993 Aug 25;21(17):4111–4118. doi: 10.1093/nar/21.17.4111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Truant R., Antunovic J., Greenblatt J., Prives C., Cromlish J. A. Direct interaction of the hepatitis B virus HBx protein with p53 leads to inhibition by HBx of p53 response element-directed transactivation. J Virol. 1995 Mar;69(3):1851–1859. doi: 10.1128/jvi.69.3.1851-1859.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Vojtek A. B., Hollenberg S. M., Cooper J. A. Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell. 1993 Jul 16;74(1):205–214. doi: 10.1016/0092-8674(93)90307-c. [DOI] [PubMed] [Google Scholar]
  35. Wang X. W., Forrester K., Yeh H., Feitelson M. A., Gu J. R., Harris C. C. Hepatitis B virus X protein inhibits p53 sequence-specific DNA binding, transcriptional activity, and association with transcription factor ERCC3. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2230–2234. doi: 10.1073/pnas.91.6.2230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Williams J. S., Andrisani O. M. The hepatitis B virus X protein targets the basic region-leucine zipper domain of CREB. Proc Natl Acad Sci U S A. 1995 Apr 25;92(9):3819–3823. doi: 10.1073/pnas.92.9.3819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Yaginuma K., Shirakata Y., Kobayashi M., Koike K. Hepatitis B virus (HBV) particles are produced in a cell culture system by transient expression of transfected HBV DNA. Proc Natl Acad Sci U S A. 1987 May;84(9):2678–2682. doi: 10.1073/pnas.84.9.2678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yen T.S.B. Hepadnaviral X Protein:Review of Recent Progress. J Biomed Sci. 1996 Jan;3(1):20–30. doi: 10.1007/BF02253575. [DOI] [PubMed] [Google Scholar]
  39. Zoulim F., Saputelli J., Seeger C. Woodchuck hepatitis virus X protein is required for viral infection in vivo. J Virol. 1994 Mar;68(3):2026–2030. doi: 10.1128/jvi.68.3.2026-2030.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]

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