Virus Genes

, Volume 5, Issue 1, pp 47–55 | Cite as

E1A gene products stimulate in vitro transcription from the adenovirus early region 4 promoter by enhancing a stable preinitiation complex

  • Takeshi Imai
  • Haruma Kawaguchi
  • Tadashi Wada
  • Hiroshi Handa


A cell-free transcription system using nuclear extracts prepared from adenovirus type 5 (Ad5)-infected and mock-infected cells were utilized to study the E1A-mediated transactivation of transcription from the Ad5 early region 4 (E4) promoter. The transcription activity of Ad-infected cell extracts was several times higher than that of mock-infected cell extracts when the E4 wild-type (WT) promoter was used. The increased activity required expression of E1A proteins. However, the transcription activities of both cell extracts were not different when the mutant promoter containing only the E4 TATA box was used. The formation of a stable preinitiation complex at the E4 WT promoter was facilitated by expression of E1A, whereas it was not facilitated at the mutant promoter. The results suggested that the E1A proteins stimulated transcription from the E4 promoter by enhancing the formation of a stable preinitiation complex through the upstream elements, together with the TATA box.

Key words

Ad E4 E1A proteins transactivation in vitro transcription preinitiation complex 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    JonesN. and ShenkT., Proc Natl Acad Sci USA76, 3665–3669, 1979.Google Scholar
  2. 2.
    BerkA.J., LeeF., HarrisonT., WilliamsJ. and SharpP.A., Cell17, 935–944, 1979.Google Scholar
  3. 3.
    NevinsJ.R., Cell26, 213–220, 1981.Google Scholar
  4. 4.
    FergusonB., JonesN., RichterJ. and RosenbergM., Science224, 1343–1346, 1984.Google Scholar
  5. 5.
    KaoT.-T. and NevinsJ.R., Mol Cell Biol3, 2058–2065, 1983.Google Scholar
  6. 6.
    NevinsJ.R., Cell29, 913–919, 1982.Google Scholar
  7. 7.
    SteinR. and ZiffE.B., Mol Cell Biol4, 2792–2801, 1984.Google Scholar
  8. 8.
    GreenM.R., TreismanR. and ManiatisT., Cell35, 137–145, 1983.Google Scholar
  9. 9.
    GaynorR.B., HillmanD. and BerkA.J., Proc Natl Acad Sci USA81, 1193–1197, 1984.Google Scholar
  10. 10.
    BerkA.J., Ann Rev Genet20, 45–79, 1986.Google Scholar
  11. 11.
    LeongK. and BerkA.J., Proc Natl Acad Sci USA83, 5844–5848, 1986.Google Scholar
  12. 12.
    SpanglerR., BrunerM., DalieB. and HarterM.L., Science237, 1044–1046, 1987.Google Scholar
  13. 13.
    GreenM., LoewensteinP.M., PusztaiR. and SymingtonJ.S., Cell53, 921–926, 1988.Google Scholar
  14. 14.
    HandaH. and SharpP.A., Mol Cell Biol4, 791–798, 1984.Google Scholar
  15. 15.
    HanakaS., NishigakiT., SharpP.A. and HandaH., Mol Cell Biol7, 2578–2587, 1987.Google Scholar
  16. 16.
    WatanabeH., ImaiT., SharpP.A. and HandaH., Mol Cell Biol8, 1290–1300, 1988.Google Scholar
  17. 17.
    NishigakiT., HanakaS., KingstonR.E. and HandaH., Mol Cell Biol8, 353–360, 1988.Google Scholar
  18. 18.
    OoyamaS., ImaiH., HanakaS. and HandaH., EMBO J8, 863–868, 1989.Google Scholar
  19. 19.
    DignamJ.D., LebovitzR.M. and RoederR.G., Nucleic Acids Res11, 1475–1489, 1983.Google Scholar
  20. 20.
    BradfordM.M., Anal Biochem72, 248–254, 1976.Google Scholar
  21. 21.
    SambrookJ., FritschE.F. and ManiatisT.,Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, 1989.Google Scholar
  22. 22.
    HawleyD.K. and RoederR.G., J Biol Chem260, 8163–8172, 1985.Google Scholar
  23. 23.
    HawleyD.K. and RoederR.G., J Biol Chem262, 3452–3461, 1987.Google Scholar
  24. 24.
    PeiR. and BerkA.J., J Virol63, 3499–3506, 1989.Google Scholar
  25. 25.
    HaiT., LiuF., AllegrettoE.A., KarinM. and GreenM.R., Gene Dev2, 1216–1226, 1988.Google Scholar
  26. 26.
    KawaguchiH., AsaiA., OhtsukaY., WatanabeH., WadaT. and HandaH., Nucleic Acids Res17, 6229–6240, 1989.Google Scholar
  27. 27.
    CortesP., BuckbinderL., LezaM.A., RakN., HearingP., MerinoA. and ReinbergD., Genes Dev2, 975–990, 1988.Google Scholar
  28. 28.
    WatanabeH., WadaT. and HandaH., EMBO J9, 841–847, 1990.Google Scholar
  29. 29.
    HorikoshiM., HaiT., LinY.-S., GreenM.R. and RoederR.G., Cell54, 1033–1042, 1988.Google Scholar
  30. 30.
    HaiT., HorikoshiM., RoederR.G. and GreenM.R., Cell54, 1043–1051, 1988.Google Scholar

Copyright information

© Kluwer Academic Publishers 1991

Authors and Affiliations

  • Takeshi Imai
    • 1
  • Haruma Kawaguchi
    • 2
  • Tadashi Wada
    • 1
  • Hiroshi Handa
    • 1
  1. 1.Department of Bacteriology, Faculty of MedicineThe University of TokyoTokyoJapan
  2. 2.Department of Applied Chemistry, Faculty of Science and TechnologyKeio UniversityHiyoshi, Yokohama 223Japan

Personalised recommendations