• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of narLink to Publisher's site
Nucleic Acids Res. Dec 1, 1996; 24(23): 4741–4750.
PMCID: PMC146319

XAP2, a novel hepatitis B virus X-associated protein that inhibits X transactivation.

Abstract

The hepatitis B virus X protein is a promiscuous transcriptional transactivator. Transactivation by the X protein is most likely mediated through binding to different cellular factors. Using the yeast two-hybrid method, we have isolated a clone that encodes a novel X-associated cellular protein: XAP2. X and XAP2 interactions also occur in vitro. Antiserum raised against XAP2 recognizes a cytoplasmic protein with an apparent molecular mass of 36 kDa. The interaction between X and XAP2 requires a small region on X containing amino acids 13-26. From Northern blot analyses, XAP2 is ubiquitously expressed in both liver-derived and non-liver-derived cell lines as well as in normal non-liver tissues. In contrast, XAP2 is expressed in very low level in the normal human liver. In transfection assays, overexpression of XAP2 abolishes transactivation by the X protein. Based on these results, we suggest that XAP2 is an important cellular negative regulator of the X protein, and that X-XAP2 interaction may play a role in HBV pathology.

Full Text

The Full Text of this article is available as a PDF (469K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Chen HS, Kaneko S, Girones R, Anderson RW, Hornbuckle WE, Tennant BC, Cote PJ, Gerin JL, Purcell RH, Miller RH. The woodchuck hepatitis virus X gene is important for establishment of virus infection in woodchucks. J Virol. 1993 Mar;67(3):1218–1226. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Blum HE, Zhang ZS, Galun E, von Weizsäcker F, Garner B, Liang TJ, Wands JR. Hepatitis B virus X protein is not central to the viral life cycle in vitro. J Virol. 1992 Feb;66(2):1223–1227. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Shirakata Y, Kawada M, Fujiki Y, Sano H, Oda M, Yaginuma K, Kobayashi M, Koike K. The X gene of hepatitis B virus induced growth stimulation and tumorigenic transformation of mouse NIH3T3 cells. Jpn J Cancer Res. 1989 Jul;80(7):617–621. [PubMed]
  • Höhne M, Schaefer S, Seifer M, Feitelson MA, Paul D, Gerlich WH. Malignant transformation of immortalized transgenic hepatocytes after transfection with hepatitis B virus DNA. EMBO J. 1990 Apr;9(4):1137–1145. [PMC free article] [PubMed]
  • Kim CM, Koike K, Saito I, Miyamura T, Jay G. HBx gene of hepatitis B virus induces liver cancer in transgenic mice. Nature. 1991 May 23;351(6324):317–320. [PubMed]
  • Gough NM. Core and E antigen synthesis in rodent cells transformed with hepatitis B virus DNA is associated with greater than genome length viral messenger RNAs. J Mol Biol. 1983 Apr 25;165(4):683–699. [PubMed]
  • Guo WT, Wang J, Tam G, Yen TS, Ou JS. Leaky transcription termination produces larger and smaller than genome size hepatitis B virus X gene transcripts. Virology. 1991 Apr;181(2):630–636. [PubMed]
  • Zelent AZ, Sells MA, Price PM, Mohamad A, Acs G, Christman JK. 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. 1987 Apr;61(4):1108–1115. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Levrero M, Jean-Jean O, Balsano C, Will H, Perricaudet M. Hepatitis B virus (HBV) X gene expression in human cells and anti-HBx antibodies detection in chronic HBV infection. Virology. 1990 Jan;174(1):299–304. [PubMed]
  • Feitelson MA, Clayton MM. X antigen/antibody markers in hepadnavirus infections. Antibodies to the X gene product(s). Gastroenterology. 1990 Aug;99(2):500–507. [PubMed]
  • Moriarty AM, Alexander H, Lerner RA, Thornton GB. Antibodies to peptides detect new hepatitis B antigen: serological correlation with hepatocellular carcinoma. Science. 1985 Jan 25;227(4685):429–433. [PubMed]
  • Vitvitski-Trépo L, Kay A, Pichoud C, Chevallier P, de Dinechin S, Shamoon BM, Mandart E, Trépo C, Galibert F. Early and frequent detection of HBxAg and/or anti-HBx in hepatitis B virus infection. Hepatology. 1990 Dec;12(6):1278–1283. [PubMed]
  • Katayama K, Hayashi N, Sasaki Y, Kasahara A, Ueda K, Fusamoto H, Sato N, Chisaka O, Matsubara K, Kamada T. Detection of hepatitis B virus X gene protein and antibody in type B chronic liver disease. Gastroenterology. 1989 Oct;97(4):990–998. [PubMed]
  • Rossner MT. Review: hepatitis B virus X-gene product: a promiscuous transcriptional activator. J Med Virol. 1992 Feb;36(2):101–117. [PubMed]
  • Aufiero B, Schneider RJ. The hepatitis B virus X-gene product trans-activates both RNA polymerase II and III promoters. EMBO J. 1990 Feb;9(2):497–504. [PMC free article] [PubMed]
  • Cross JC, Wen P, Rutter WJ. 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. [PMC free article] [PubMed]
  • Kekulé AS, Lauer U, Weiss L, Luber B, Hofschneider PH. Hepatitis B virus transactivator HBx uses a tumour promoter signalling pathway. Nature. 1993 Feb 25;361(6414):742–745. [PubMed]
  • Lucito R, Schneider RJ. Hepatitis B virus X protein activates transcription factor NF-kappa B without a requirement for protein kinase C. J Virol. 1992 Feb;66(2):983–991. [PMC free article] [PubMed]
  • Takada S, Kido H, Fukutomi A, Mori T, Koike K. Interaction of hepatitis B virus X protein with a serine protease, tryptase TL2 as an inhibitor. Oncogene. 1994 Feb;9(2):341–348. [PubMed]
  • Haviv I, Vaizel D, Shaul Y. The X protein of hepatitis B virus coactivates potent activation domains. Mol Cell Biol. 1995 Feb;15(2):1079–1085. [PMC free article] [PubMed]
  • Maguire HF, Hoeffler JP, 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. [PubMed]
  • Seto E, Mitchell PJ, Yen TS. Transactivation by the hepatitis B virus X protein depends on AP-2 and other transcription factors. Nature. 1990 Mar 1;344(6261):72–74. [PubMed]
  • Unger T, Shaul Y. The X protein of the hepatitis B virus acts as a transcription factor when targeted to its responsive element. EMBO J. 1990 Jun;9(6):1889–1895. [PMC free article] [PubMed]
  • Seto E, Zhou DX, Peterlin BM, Yen TS. trans-activation by the hepatitis B virus X protein shows cell-type specificity. Virology. 1989 Dec;173(2):764–766. [PubMed]
  • Jameel S, Siddiqui A, Maguire HF, Rao KV. Hepatitis B virus X protein produced in Escherichia coli is biologically functional. J Virol. 1990 Aug;64(8):3963–3966. [PMC free article] [PubMed]
  • Ritter SE, Whitten TM, Quets AT, Schloemer RH. An internal domain of the hepatitis B virus X antigen is necessary for transactivating activity. Virology. 1991 Jun;182(2):841–845. [PubMed]
  • Twu JS, Wu JY, Robinson WS. Transcriptional activation of the human immunodeficiency virus type 1 long terminal repeat by hepatitis B virus X-protein requires de novo protein synthesis. Virology. 1990 Jul;177(1):406–410. [PubMed]
  • Cheong JH, 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. [PMC free article] [PubMed]
  • Feitelson MA, Zhu M, Duan LX, London WT. 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]
  • Qadri I, Maguire HF, Siddiqui A. Hepatitis B virus transactivator protein X interacts with the TATA-binding protein. Proc Natl Acad Sci U S A. 1995 Feb 14;92(4):1003–1007. [PMC free article] [PubMed]
  • Seto E, Yen TS. Mutual functional antagonism of the simian virus 40 T antigen and the hepatitis B virus trans activator. J Virol. 1991 May;65(5):2351–2356. [PMC free article] [PubMed]
  • Lee TH, Elledge SJ, Butel JS. Hepatitis B virus X protein interacts with a probable cellular DNA repair protein. J Virol. 1995 Feb;69(2):1107–1114. [PMC free article] [PubMed]
  • Elledge SJ, Mulligan JT, Ramer SW, Spottswood M, Davis RW. Lambda YES: a multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations. Proc Natl Acad Sci U S A. 1991 Mar 1;88(5):1731–1735. [PMC free article] [PubMed]
  • Durfee T, Becherer K, Chen PL, Yeh SH, Yang Y, Kilburn AE, Lee WH, Elledge SJ. The retinoblastoma protein associates with the protein phosphatase type 1 catalytic subunit. Genes Dev. 1993 Apr;7(4):555–569. [PubMed]
  • Inouye CJ, Seto E. Relief of YY1-induced transcriptional repression by protein-protein interaction with the nucleolar phosphoprotein B23. J Biol Chem. 1994 Mar 4;269(9):6506–6510. [PubMed]
  • Ron D, Dressler H. pGSTag--a versatile bacterial expression plasmid for enzymatic labeling of recombinant proteins. Biotechniques. 1992 Dec;13(6):866–869. [PubMed]
  • Luban J, Bossolt KL, Franke EK, Kalpana GV, Goff SP. Human immunodeficiency virus type 1 Gag protein binds to cyclophilins A and B. Cell. 1993 Jun 18;73(6):1067–1078. [PubMed]
  • Seto E, Yen TS, Peterlin BM, Ou JH. Trans-activation of the human immunodeficiency virus long terminal repeat by the hepatitis B virus X protein. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8286–8290. [PMC free article] [PubMed]
  • Gorman CM, Merlino GT, Willingham MC, Pastan I, Howard BH. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. [PMC free article] [PubMed]
  • Gorman CM, Moffat LF, Howard BH. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. [PMC free article] [PubMed]
  • Peterlin BM, Luciw PA, Barr PJ, Walker MD. Elevated levels of mRNA can account for the trans-activation of human immunodeficiency virus. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9734–9738. [PMC free article] [PubMed]
  • Lillie JW, Green MR. Transcription activation by the adenovirus E1a protein. Nature. 1989 Mar 2;338(6210):39–44. [PubMed]
  • Sadowski I, Ma J, Triezenberg S, Ptashne M. GAL4-VP16 is an unusually potent transcriptional activator. Nature. 1988 Oct 6;335(6190):563–564. [PubMed]
  • Breeden L, Nasmyth K. Regulation of the yeast HO gene. Cold Spring Harb Symp Quant Biol. 1985;50:643–650. [PubMed]
  • Becker DM, Guarente L. High-efficiency transformation of yeast by electroporation. Methods Enzymol. 1991;194:182–187. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PMC free article] [PubMed]
  • Guarente L. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol. 1983;101:181–191. [PubMed]
  • Frangioni JV, Neel BG. Solubilization and purification of enzymatically active glutathione S-transferase (pGEX) fusion proteins. Anal Biochem. 1993 Apr;210(1):179–187. [PubMed]
  • Oettinger HF, Pasqualini R, Bernfield M. Recombinant peptides as immunogens: a comparison of protocols for antisera production using the pGEX system. Biotechniques. 1992 Apr;12(4):544–549. [PubMed]
  • Graham FL, van der Eb AJ. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. [PubMed]
  • Chien CT, Bartel PL, Sternglanz R, Fields S. The two-hybrid system: a method to identify and clone genes for proteins that interact with a protein of interest. Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9578–9582. [PMC free article] [PubMed]
  • Fields S, Song O. A novel genetic system to detect protein-protein interactions. Nature. 1989 Jul 20;340(6230):245–246. [PubMed]
  • Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. [PMC free article] [PubMed]
  • Takada S, Koike K. Three sites of the hepatitis B virus X protein cooperatively interact with cellular proteins. Virology. 1994 Dec;205(2):503–510. [PubMed]
  • Lee TH, Finegold MJ, Shen RF, DeMayo JL, Woo SL, Butel JS. Hepatitis B virus transactivator X protein is not tumorigenic in transgenic mice. J Virol. 1990 Dec;64(12):5939–5947. [PMC free article] [PubMed]
  • Zheng YW, Riegler J, Wu J, Yen TS. Novel short transcripts of hepatitis B virus X gene derived from intragenic promoter. J Biol Chem. 1994 Sep 9;269(36):22593–22598. [PubMed]
  • Seifer M, Heermann KH, Gerlich WH. Expression pattern of the hepatitis B virus genome in transfected mouse fibroblasts. Virology. 1990 Nov;179(1):287–299. [PubMed]
  • 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. [PubMed]
  • Schek N, Bartenschlager R, Kuhn C, Schaller H. Phosphorylation and rapid turnover of hepatitis B virus X-protein expressed in HepG2 cells from a recombinant vaccinia virus. Oncogene. 1991 Oct;6(10):1735–1744. [PubMed]
  • 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. [PMC free article] [PubMed]
  • Doria M, Klein N, Lucito R, Schneider RJ. 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. [PMC free article] [PubMed]
  • Hu KQ, Vierling JM, Siddiqui A. Trans-activation of HLA-DR gene by hepatitis B virus X gene product. Proc Natl Acad Sci U S A. 1990 Sep;87(18):7140–7144. [PMC free article] [PubMed]
  • Murakami S, Cheong JH, Kaneko S. Human hepatitis virus X gene encodes a regulatory domain that represses transactivation of X protein. J Biol Chem. 1994 May 27;269(21):15118–15123. [PubMed]
  • Siddiqui A, Gaynor R, Srinivasan A, Mapoles J, Farr RW. trans-activation of viral enhancers including long terminal repeat of the human immunodeficiency virus by the hepatitis B virus X protein. Virology. 1989 Apr;169(2):479–484. [PubMed]
  • Spandau DF, Lee CH. trans-activation of viral enhancers by the hepatitis B virus X protein. J Virol. 1988 Feb;62(2):427–434. [PMC free article] [PubMed]
  • Takada S, Koike K. Trans-activation function of a 3' truncated X gene-cell fusion product from integrated hepatitis B virus DNA in chronic hepatitis tissues. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5628–5632. [PMC free article] [PubMed]
  • Twu JS, Robinson WS. Hepatitis B virus X gene can transactivate heterologous viral sequences. Proc Natl Acad Sci U S A. 1989 Mar;86(6):2046–2050. [PMC free article] [PubMed]
  • Zahm P, Hofschneider PH, Koshy R. The HBV X-ORF encodes a transactivator: a potential factor in viral hepatocarcinogenesis. Oncogene. 1988 Aug;3(2):169–177. [PubMed]
  • Faktor O, Shaul Y. The identification of hepatitis B virus X gene responsive elements reveals functional similarity of X and HTLV-I tax. Oncogene. 1990 Jun;5(6):867–872. [PubMed]
  • 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]
  • Lin MH, Lo SC. Dimerization of hepatitis B viral X protein synthesized in a cell-free system. Biochem Biophys Res Commun. 1989 Oct 16;164(1):14–21. [PubMed]
  • Momand J, Zambetti GP, Olson DC, George D, Levine AJ. The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation. Cell. 1992 Jun 26;69(7):1237–1245. [PubMed]
  • Oliner JD, Pietenpol JA, Thiagalingam S, Gyuris J, Kinzler KW, Vogelstein B. Oncoprotein MDM2 conceals the activation domain of tumour suppressor p53. Nature. 1993 Apr 29;362(6423):857–860. [PubMed]
  • Benezra R, Davis RL, Lockshon D, Turner DL, Weintraub H. The protein Id: a negative regulator of helix-loop-helix DNA binding proteins. Cell. 1990 Apr 6;61(1):49–59. [PubMed]
  • Cordle SR, Henderson E, Masuoka H, Weil PA, Stein R. Pancreatic beta-cell-type-specific transcription of the insulin gene is mediated by basic helix-loop-helix DNA-binding proteins. Mol Cell Biol. 1991 Mar;11(3):1734–1738. [PMC free article] [PubMed]
  • Jen Y, Weintraub H, Benezra R. Overexpression of Id protein inhibits the muscle differentiation program: in vivo association of Id with E2A proteins. Genes Dev. 1992 Aug;6(8):1466–1479. [PubMed]
  • Kreider BL, Benezra R, Rovera G, Kadesch T. Inhibition of myeloid differentiation by the helix-loop-helix protein Id. Science. 1992 Mar 27;255(5052):1700–1702. [PubMed]
  • Wilson RB, Kiledjian M, Shen CP, Benezra R, Zwollo P, Dymecki SM, Desiderio SV, Kadesch T. Repression of immunoglobulin enhancers by the helix-loop-helix protein Id: implications for B-lymphoid-cell development. Mol Cell Biol. 1991 Dec;11(12):6185–6191. [PMC free article] [PubMed]
  • Beijersbergen RL, Hijmans EM, Zhu L, Bernards R. Interaction of c-Myc with the pRb-related protein p107 results in inhibition of c-Myc-mediated transactivation. EMBO J. 1994 Sep 1;13(17):4080–4086. [PMC free article] [PubMed]
  • Hateboer G, Gennissen A, Ramos YF, Kerkhoven RM, Sonntag-Buck V, Stunnenberg HG, Bernards R. BS69, a novel adenovirus E1A-associated protein that inhibits E1A transactivation. EMBO J. 1995 Jul 3;14(13):3159–3169. [PMC free article] [PubMed]
  • Kovesdi I, Reichel R, Nevins JR. Identification of a cellular transcription factor involved in E1A trans-activation. Cell. 1986 Apr 25;45(2):219–228. [PubMed]
  • Bagchi S, Weinmann R, Raychaudhuri P. The retinoblastoma protein copurifies with E2F-I, an E1A-regulated inhibitor of the transcription factor E2F. Cell. 1991 Jun 14;65(6):1063–1072. [PubMed]
  • Chellappan SP, Hiebert S, Mudryj M, Horowitz JM, Nevins JR. The E2F transcription factor is a cellular target for the RB protein. Cell. 1991 Jun 14;65(6):1053–1061. [PubMed]
  • Chittenden T, Livingston DM, Kaelin WG., Jr The T/E1A-binding domain of the retinoblastoma product can interact selectively with a sequence-specific DNA-binding protein. Cell. 1991 Jun 14;65(6):1073–1082. [PubMed]
  • Baeuerle PA, Baltimore D. Activation of DNA-binding activity in an apparently cytoplasmic precursor of the NF-kappa B transcription factor. Cell. 1988 Apr 22;53(2):211–217. [PubMed]
  • Baeuerle PA, Baltimore D. I kappa B: a specific inhibitor of the NF-kappa B transcription factor. Science. 1988 Oct 28;242(4878):540–546. [PubMed]
  • Ganem D, Varmus HE. The molecular biology of the hepatitis B viruses. Annu Rev Biochem. 1987;56:651–693. [PubMed]
  • Acs G, Sells MA, Purcell RH, Price P, Engle R, Shapiro M, Popper H. Hepatitis B virus produced by transfected Hep G2 cells causes hepatitis in chimpanzees. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4641–4644. [PMC free article] [PubMed]
  • Sureau C, Romet-Lemonne JL, Mullins JI, Essex M. Production of hepatitis B virus by a differentiated human hepatoma cell line after transfection with cloned circular HBV DNA. Cell. 1986 Oct 10;47(1):37–47. [PubMed]
  • Tsurimoto T, Fujiyama A, Matsubara K. Stable expression and replication of hepatitis B virus genome in an integrated state in a human hepatoma cell line transfected with the cloned viral DNA. Proc Natl Acad Sci U S A. 1987 Jan;84(2):444–448. [PMC free article] [PubMed]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

  • Cited in Books
    Cited in Books
    PubMed Central articles cited in books
  • Gene
    Gene
    Gene links
  • Gene (nucleotide)
    Gene (nucleotide)
    Records in Gene identified from shared sequence links
  • GEO Profiles
    GEO Profiles
    Related GEO records
  • HomoloGene
    HomoloGene
    HomoloGene links
  • MedGen
    MedGen
    Related information in MedGen
  • Nucleotide
    Nucleotide
    Published Nucleotide sequences
  • OMIM
    OMIM
    OMIM record citing PubMed
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links
  • Taxonomy
    Taxonomy
    Related taxonomy entry
  • Taxonomy Tree
    Taxonomy Tree

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...