Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1997 Jul;71(7):5102–5109. doi: 10.1128/jvi.71.7.5102-5109.1997

The product of the adenovirus intermediate gene IX is a transcriptional activator.

P Lutz 1, M Rosa-Calatrava 1, C Kedinger 1
PMCID: PMC191744  PMID: 9188576

Abstract

We have investigated the functional properties of the product of the adenovirus type 5 gene IX. This gene, which is expressed at intermediate times postinfection, encodes a small polypeptide (pIX) of 140 residues that has previously been shown to be incorporated into the viral capsid. Here, we show that pIX, in addition to its structural contribution, exhibits transcriptional properties. In transient transfection experiments, expression of pIX stimulated adenovirus major late promoter activity. The effect was independent of other viral proteins, but the level of promoter activation appeared strongly pIX dose dependent; similar levels of induction were observed with other cellular or viral TATA-containing (but not with TATA-less) promoters. This promoter specificity could be reproduced in a cell-free transcription system by the addition of purified recombinant pIX, further stressing the transcriptional nature of the phenomenon. A preliminary structural analysis of pIX indicated that the integrity of a putative leucine zipper at the carboxy-terminal end of the molecule, as well as elements within the amino-terminal half, was critical for pIX transcriptional activity. The relevance of these findings in adenovirus infection is discussed.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

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

  1. Adya N., Giam C. Z. Distinct regions in human T-cell lymphotropic virus type I tax mediate interactions with activator protein CREB and basal transcription factors. J Virol. 1995 Mar;69(3):1834–1841. doi: 10.1128/jvi.69.3.1834-1841.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Akusjärvi G. Proteins with transcription regulatory properties encoded by human adenoviruses. Trends Microbiol. 1993 Aug;1(5):163–170. doi: 10.1016/0966-842x(93)90085-6. [DOI] [PubMed] [Google Scholar]
  3. Aleström P., Akusjärvi G., Perricaudet M., Mathews M. B., Klessig D. F., Pettersson U. The gene for polypeptide IX of adenovirus type 2 and its unspliced messenger RNA. Cell. 1980 Mar;19(3):671–681. doi: 10.1016/s0092-8674(80)80044-4. [DOI] [PubMed] [Google Scholar]
  4. Ali S., Lutz Y., Bellocq J. P., Chenard-Neu M. P., Rouyer N., Metzger D. Production and characterization of monoclonal antibodies recognising defined regions of the human oestrogen receptor. Hybridoma. 1993 Aug;12(4):391–405. doi: 10.1089/hyb.1993.12.391. [DOI] [PubMed] [Google Scholar]
  5. Alonso-Caplen F. V., Katze M. G., Krug R. M. Efficient transcription, not translation, is dependent on adenovirus tripartite leader sequences at late times of infection. J Virol. 1988 May;62(5):1606–1616. doi: 10.1128/jvi.62.5.1606-1616.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bedrosian C. L., Bastia D. The DNA-binding domain of HPV-16 E2 protein interaction with the viral enhancer: protein-induced DNA bending and role of the nonconserved core sequence in binding site affinity. Virology. 1990 Feb;174(2):557–575. doi: 10.1016/0042-6822(90)90109-5. [DOI] [PubMed] [Google Scholar]
  7. Berk A. J., Lee F., Harrison T., Williams J., Sharp P. A. Pre-early adenovirus 5 gene product regulates synthesis of early viral messenger RNAs. Cell. 1979 Aug;17(4):935–944. doi: 10.1016/0092-8674(79)90333-7. [DOI] [PubMed] [Google Scholar]
  8. Bocco J. L., Reimund B., Chatton B., Kedinger C. Rb may act as a transcriptional co-activator in undifferentiated F9 cells. Oncogene. 1993 Nov;8(11):2977–2986. [PubMed] [Google Scholar]
  9. Boshart M., Klüppel M., Schmidt A., Schütz G., Luckow B. Reporter constructs with low background activity utilizing the cat gene. Gene. 1992 Jan 2;110(1):129–130. doi: 10.1016/0378-1119(92)90456-y. [DOI] [PubMed] [Google Scholar]
  10. Boulanger P., Lemay P., Blair G. E., Russell W. C. Characterization of adenovirus protein IX. J Gen Virol. 1979 Sep;44(3):783–800. doi: 10.1099/0022-1317-44-3-783. [DOI] [PubMed] [Google Scholar]
  11. Boulikas T. Putative nuclear localization signals (NLS) in protein transcription factors. J Cell Biochem. 1994 May;55(1):32–58. doi: 10.1002/jcb.240550106. [DOI] [PubMed] [Google Scholar]
  12. Burke T. W., Kadonaga J. T. Drosophila TFIID binds to a conserved downstream basal promoter element that is present in many TATA-box-deficient promoters. Genes Dev. 1996 Mar 15;10(6):711–724. doi: 10.1101/gad.10.6.711. [DOI] [PubMed] [Google Scholar]
  13. Burley S. K., Roeder R. G. Biochemistry and structural biology of transcription factor IID (TFIID). Annu Rev Biochem. 1996;65:769–799. doi: 10.1146/annurev.bi.65.070196.004005. [DOI] [PubMed] [Google Scholar]
  14. Caravokyri C., Leppard K. N. Constitutive episomal expression of polypeptide IX (pIX) in a 293-based cell line complements the deficiency of pIX mutant adenovirus type 5. J Virol. 1995 Nov;69(11):6627–6633. doi: 10.1128/jvi.69.11.6627-6633.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Chao D. M., Gadbois E. L., Murray P. J., Anderson S. F., Sonu M. S., Parvin J. D., Young R. A. A mammalian SRB protein associated with an RNA polymerase II holoenzyme. Nature. 1996 Mar 7;380(6569):82–85. doi: 10.1038/380082a0. [DOI] [PubMed] [Google Scholar]
  16. Chatton B., Bocco J. L., Gaire M., Hauss C., Reimund B., Goetz J., Kedinger C. Transcriptional activation by the adenovirus larger E1a product is mediated by members of the cellular transcription factor ATF family which can directly associate with E1a. Mol Cell Biol. 1993 Jan;13(1):561–570. doi: 10.1128/mcb.13.1.561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Chen C., Okayama H. High-efficiency transformation of mammalian cells by plasmid DNA. Mol Cell Biol. 1987 Aug;7(8):2745–2752. doi: 10.1128/mcb.7.8.2745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Colby W. W., Shenk T. Adenovirus type 5 virions can be assembled in vivo in the absence of detectable polypeptide IX. J Virol. 1981 Sep;39(3):977–980. doi: 10.1128/jvi.39.3.977-980.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Cook W. J., Gu B., DeLuca N. A., Moynihan E. B., Coen D. M. Induction of transcription by a viral regulatory protein depends on the relative strengths of functional TATA boxes. Mol Cell Biol. 1995 Sep;15(9):4998–5006. doi: 10.1128/mcb.15.9.4998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Damania B., Alwine J. C. TAF-like function of SV40 large T antigen. Genes Dev. 1996 Jun 1;10(11):1369–1381. doi: 10.1101/gad.10.11.1369. [DOI] [PubMed] [Google Scholar]
  21. Dingwall C., Laskey R. A. Nuclear targeting sequences--a consensus? Trends Biochem Sci. 1991 Dec;16(12):478–481. doi: 10.1016/0968-0004(91)90184-w. [DOI] [PubMed] [Google Scholar]
  22. Dynlacht B. D., Hoey T., Tjian R. Isolation of coactivators associated with the TATA-binding protein that mediate transcriptional activation. Cell. 1991 Aug 9;66(3):563–576. doi: 10.1016/0092-8674(81)90019-2. [DOI] [PubMed] [Google Scholar]
  23. Flint S. J. Expression of adenoviral genetic information in productively infected cells. Biochim Biophys Acta. 1982 Apr 29;651(2-3):175–208. doi: 10.1016/0304-419x(82)90011-7. [DOI] [PubMed] [Google Scholar]
  24. Garcia-Carranca A., Thierry F., Yaniv M. Interplay of viral and cellular proteins along the long control region of human papillomavirus type 18. J Virol. 1988 Nov;62(11):4321–4330. doi: 10.1128/jvi.62.11.4321-4330.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Gerster T., Roeder R. G. A herpesvirus trans-activating protein interacts with transcription factor OTF-1 and other cellular proteins. Proc Natl Acad Sci U S A. 1988 Sep;85(17):6347–6351. doi: 10.1073/pnas.85.17.6347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Gilardi P., Perricaudet M. The E4 promoter of adenovirus type 2 contains an E1A dependent cis-acting element. Nucleic Acids Res. 1986 Nov 25;14(22):9035–9049. doi: 10.1093/nar/14.22.9035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Goetz J., Chatton B., Mattei M. G., Kedinger C. Structure and expression of the ATFa gene. J Biol Chem. 1996 Nov 22;271(47):29589–29598. doi: 10.1074/jbc.271.47.29589. [DOI] [PubMed] [Google Scholar]
  28. Gopalkrishnan R. V., Dollé P., Mattei M. G., La Thangue N. B., Kedinger C. Genomic structure and developmental expression of the mouse cell cycle regulatory transcription factor DP1. Oncogene. 1996 Dec 19;13(12):2671–2680. [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Horikoshi N., Maguire K., Kralli A., Maldonado E., Reinberg D., Weinmann R. Direct interaction between adenovirus E1A protein and the TATA box binding transcription factor IID. Proc Natl Acad Sci U S A. 1991 Jun 15;88(12):5124–5128. doi: 10.1073/pnas.88.12.5124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Imperiale M. J., Nevins J. R. Adenovirus 5 E2 transcription unit: an E1A-inducible promoter with an essential element that functions independently of position or orientation. Mol Cell Biol. 1984 May;4(5):875–882. doi: 10.1128/mcb.4.5.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Jansen-Durr P., Boeuf H., Kédinger C. Replication-induced stimulation of the major late promoter of adenovirus is correlated to the binding of a factor to sequences in the first intron. Nucleic Acids Res. 1988 May 11;16(9):3771–3786. doi: 10.1093/nar/16.9.3771. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Jones N., Shenk T. Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells. Cell. 1979 Jul;17(3):683–689. doi: 10.1016/0092-8674(79)90275-7. [DOI] [PubMed] [Google Scholar]
  35. Kim Y. J., Björklund S., Li Y., Sayre M. H., Kornberg R. D. A multiprotein mediator of transcriptional activation and its interaction with the C-terminal repeat domain of RNA polymerase II. Cell. 1994 May 20;77(4):599–608. doi: 10.1016/0092-8674(94)90221-6. [DOI] [PubMed] [Google Scholar]
  36. Koleske A. J., Young R. A. An RNA polymerase II holoenzyme responsive to activators. Nature. 1994 Mar 31;368(6470):466–469. doi: 10.1038/368466a0. [DOI] [PubMed] [Google Scholar]
  37. Lee W. S., Kao C. C., Bryant G. O., Liu X., Berk A. J. Adenovirus E1A activation domain binds the basic repeat in the TATA box transcription factor. Cell. 1991 Oct 18;67(2):365–376. doi: 10.1016/0092-8674(91)90188-5. [DOI] [PubMed] [Google Scholar]
  38. Leong K., Berk A. J. Adenovirus early region 1A protein increases the number of template molecules transcribed in cell-free extracts. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5844–5848. doi: 10.1073/pnas.83.16.5844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Leong K., Lee W., Berk A. J. High-level transcription from the adenovirus major late promoter requires downstream binding sites for late-phase-specific factors. J Virol. 1990 Jan;64(1):51–60. doi: 10.1128/jvi.64.1.51-60.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Liu F., Green M. R. A specific member of the ATF transcription factor family can mediate transcription activation by the adenovirus E1a protein. Cell. 1990 Jun 29;61(7):1217–1224. doi: 10.1016/0092-8674(90)90686-9. [DOI] [PubMed] [Google Scholar]
  41. Lutz P., Kedinger C. Properties of the adenovirus IVa2 gene product, an effector of late-phase-dependent activation of the major late promoter. J Virol. 1996 Mar;70(3):1396–1405. doi: 10.1128/jvi.70.3.1396-1405.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Lutz P., Puvion-Dutilleul F., Lutz Y., Kedinger C. Nucleoplasmic and nucleolar distribution of the adenovirus IVa2 gene product. J Virol. 1996 Jun;70(6):3449–3460. doi: 10.1128/jvi.70.6.3449-3460.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Mason B. B., Davis A. R., Bhat B. M., Chengalvala M., Lubeck M. D., Zandle G., Kostek B., Cholodofsky S., Dheer S., Molnar-Kimber K. Adenovirus vaccine vectors expressing hepatitis B surface antigen: importance of regulatory elements in the adenovirus major late intron. Virology. 1990 Aug;177(2):452–461. doi: 10.1016/0042-6822(90)90509-p. [DOI] [PubMed] [Google Scholar]
  44. Matthews M. A., Markowitz R. B., Dynan W. S. In vitro activation of transcription by the human T-cell leukemia virus type I Tax protein. Mol Cell Biol. 1992 May;12(5):1986–1996. doi: 10.1128/mcb.12.5.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. McBride A. A., Schlegel R., Howley P. M. The carboxy-terminal domain shared by the bovine papillomavirus E2 transactivator and repressor proteins contains a specific DNA binding activity. EMBO J. 1988 Feb;7(2):533–539. doi: 10.1002/j.1460-2075.1988.tb02842.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. McKnight S. L., Gavis E. R., Kingsbury R., Axel R. Analysis of transcriptional regulatory signals of the HSV thymidine kinase gene: identification of an upstream control region. Cell. 1981 Aug;25(2):385–398. doi: 10.1016/0092-8674(81)90057-x. [DOI] [PubMed] [Google Scholar]
  47. Meisterernst M., Roy A. L., Lieu H. M., Roeder R. G. Activation of class II gene transcription by regulatory factors is potentiated by a novel activity. Cell. 1991 Sep 6;66(5):981–993. doi: 10.1016/0092-8674(91)90443-3. [DOI] [PubMed] [Google Scholar]
  48. Mondésert G., Kédinger C. Cooperation between upstream and downstream elements of the adenovirus major late promoter for maximal late phase-specific transcription. Nucleic Acids Res. 1991 Jun 25;19(12):3221–3228. doi: 10.1093/nar/19.12.3221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Nevins J. R. Transcriptional activation by viral regulatory proteins. Trends Biochem Sci. 1991 Nov;16(11):435–439. doi: 10.1016/0968-0004(91)90171-q. [DOI] [PubMed] [Google Scholar]
  50. Qadri I., Conaway J. W., Conaway R. C., Schaack J., Siddiqui A. Hepatitis B virus transactivator protein, HBx, associates with the components of TFIIH and stimulates the DNA helicase activity of TFIIH. Proc Natl Acad Sci U S A. 1996 Oct 1;93(20):10578–10583. doi: 10.1073/pnas.93.20.10578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Sayre M. H., Tschochner H., Kornberg R. D. Reconstitution of transcription with five purified initiation factors and RNA polymerase II from Saccharomyces cerevisiae. J Biol Chem. 1992 Nov 15;267(32):23376–23382. [PubMed] [Google Scholar]
  52. Stewart P. L., Fuller S. D., Burnett R. M. Difference imaging of adenovirus: bridging the resolution gap between X-ray crystallography and electron microscopy. EMBO J. 1993 Jul;12(7):2589–2599. doi: 10.1002/j.1460-2075.1993.tb05919.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  54. Takayesu D., Teodoro J. G., Whalen S. G., Branton P. E. Characterization of the 55K adenovirus type 5 E1B product and related proteins. J Gen Virol. 1994 Apr;75(Pt 4):789–798. doi: 10.1099/0022-1317-75-4-789. [DOI] [PubMed] [Google Scholar]
  55. Tanese N., Pugh B. F., Tjian R. Coactivators for a proline-rich activator purified from the multisubunit human TFIID complex. Genes Dev. 1991 Dec;5(12A):2212–2224. doi: 10.1101/gad.5.12a.2212. [DOI] [PubMed] [Google Scholar]
  56. Tribouley C., Lutz P., Staub A., Kedinger C. The product of the adenovirus intermediate gene IVa2 is a transcriptional activator of the major late promoter. J Virol. 1994 Jul;68(7):4450–4457. doi: 10.1128/jvi.68.7.4450-4457.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Webster N. J., Green S., Tasset D., Ponglikitmongkol M., Chambon P. The transcriptional activation function located in the hormone-binding domain of the human oestrogen receptor is not encoded in a single exon. EMBO J. 1989 May;8(5):1441–1446. doi: 10.1002/j.1460-2075.1989.tb03526.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Webster N., Jin J. R., Green S., Hollis M., Chambon P. The yeast UASG is a transcriptional enhancer in human HeLa cells in the presence of the GAL4 trans-activator. Cell. 1988 Jan 29;52(2):169–178. doi: 10.1016/0092-8674(88)90505-3. [DOI] [PubMed] [Google Scholar]
  59. van Oostrum J., Burnett R. M. Molecular composition of the adenovirus type 2 virion. J Virol. 1985 Nov;56(2):439–448. doi: 10.1128/jvi.56.2.439-448.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. van Ooyen A., van den Berg J., Mantei N., Weissmann C. Comparison of total sequence of a cloned rabbit beta-globin gene and its flanking regions with a homologous mouse sequence. Science. 1979 Oct 19;206(4416):337–344. doi: 10.1126/science.482942. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES