1887

Abstract

Transcription of the gene of the highly oncogenic adenovirus 12 (Ad12) initiates at two start sites (TS1 and TS2). We have previously shown that the E2F and ATF motifs distal of TS1 co-operatively participate in E1A autostimulation from the TS1 promoter region. Here we report the identification of a second E2F-like target region (E2FII) immediately upstream of the E1A-stimulating factor 1 binding site (ESF-1), important for 13S-mediated autoactivation from TS2. Reporter constructs lacking distinct TS2 c/s-acting elements were analysed for their levels of CAT expression in the absence and presence of the E1A 13S protein in transient expression assays. In the absence of 13S, full promoter activity was observed only for a construct containing all elements (the E2F-like motif, an E-Box and the TATA element). Promoter activation increased significantly in Ad12 E1A-co-transfected cells. Induction by the 13S protein was also detected for the construct containing a non-functional ESF-1 sequence. Our results indicate that the E2F-like motif is responsible for activation mediated by the 13S protein from TS2, while ESF-1-or TATA-binding protein activity were not involved. Additionally, the TATA sequence appeared to be dispensable for transactivation. Gel-shift experiments using the E2F-like promoter element as a probe indicated the binding of an E2F-5 or E2F-5-like transcription factor to this region. We conclude that transcription through the TS1 as well astheTS2 promoter region is stimulated by the Ad12 13S protein. Moreover, transfection of the construct including bothTS1 and TS2 indicates an E2F-site-mediated synergism between both regions with respect to E1A-induced transactivation.

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1997-04-01
2023-01-29
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References

  1. Bagchi S., Raychaudhuri P., Nevins J. R. 1989; Phosphorylation- dependent activation of the adenovirus-inducible E2F transcription factor in a cell-free system. Proceedings of the National Academy of Sciences, USA 86:4352–4356
    [Google Scholar]
  2. Bagchi S., Raychaudhuri P., Nevins J. R. 1990; Adenovirus E1A proteins can dissociate heteromeric complexes involving the E2F transcription factor: a novel mechanism for ElA trans-activation. Cell 62:659–669
    [Google Scholar]
  3. Barbeau D., Charbonneau X. X., Whalen S. G., Bayley S. T., Branton P. E. 1994; Functional interactions within adenovirus ElA protein complexes. Oncogene 9:359–373
    [Google Scholar]
  4. Barrett P., Clark L., Hay R. T. 1987; A cellular protein binds to a conserved sequence in the adenovirus type 2 enhancer. Nucleic Acids Research 15:2719–2735
    [Google Scholar]
  5. Beijersbergen R. L., Kerkhoven R., Zhu L., Cerlee L., Voorhoeve P. M., Bernards R. 1994; E2F-4, a new member of the E2F gene family, has oncogenic activity and associates with p107 in vivo. Genes & Development 8:2680–2690
    [Google Scholar]
  6. Berk A. J. 1986; Adenovirus promoters and ElA transactivation. Annual Review of Genetics 20:45–79
    [Google Scholar]
  7. Berk A. J., Lee F., Harrison T., Williams J., Sharp P. 1979; Preearly adenovirus 5 gene product regulates synthesis of early viral messenger RNAs. Cell 17:935–944
    [Google Scholar]
  8. Blake M. C., Azizkhan J. C. 1989; Transcription factor E2F is required for efficient expression of the hamster dihydrofolate reductase gene in vitro and in vivo. Molecular and Cellular Biology 11:4994–5002
    [Google Scholar]
  9. Boyer T. G., Berk A. J. 1993; Functional interactions of the adenovirus ElA with holo-TFIID. Genes & Development 7:1810–1823
    [Google Scholar]
  10. Branton P. E., Bayley S. T., Graham F. L. 1985; Transformation by human adenoviruses. Biochimica et Biophysica Acta 780:67–94
    [Google Scholar]
  11. Brockmann D., Bury C., Kroener G., Kirch H. C., Esche H. 1995; Repression of the c-jun trans-activation function by the adenovirus type 12 E1A 52R protein correlates with the inhibition of phosphorylation of the c-jun activation domain. Journal of Biological Chemistry 270:10754–10763
    [Google Scholar]
  12. Buck V., Allen K. E., Sorensen T., Bybee A., Hijmans E. M., Voorhoeve P. M., Bernards R., La Thangue N. B. 1995; Molecular and functional characterisation of E2F-5, a new member of the E2F family. Oncogene 11:31–38
    [Google Scholar]
  13. Chatton B., Bocco J. L., Gaire M., Hauss C., Reimund B., Goetz J., Kedinger C. 1993; 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. Molecular and Cellular Biology 13:561–570
    [Google Scholar]
  14. Cobrinik D., Whyte P., Peeper D. S., Jacks T., Weinberg R. A. 1993; Cell cycle-specific association of E2F with the p130 E1A-binding protein. Genes & Development 7:2392–2404
    [Google Scholar]
  15. Cogan J. D., Jones S. N., Hall R. K., Tibbetts C. 1992; Functional diversity of E1A gene autoregulation among human adenoviruses. Journal of Virology 66:3833–3845
    [Google Scholar]
  16. Dalton S. 1992; Cell cycle regulation of the human cdc2 gene. EMBO Journal 11:1797–1804
    [Google Scholar]
  17. De Groot R., Foulkes N., Mulder M., Kruijer W., Sassone-Corsi P. 1991; Positive regulation of jun/AP1 by E1a. Molecular and Cellular Biology 11:192–201
    [Google Scholar]
  18. Dynlacht B. D., Hoey T., Tjian R. 1991; Isolation of co-activators associated with the TATA-binding protein that mediate transcriptional activation. Cell 66:563–576
    [Google Scholar]
  19. Fujinaga K., Yoshida K., Yamashita T., Shimizu Y. 1984; Organization, integration and transcription of transforming genes of oncogenic human adenovirus types 12 and 7. Current Topics in Microbiology and Immunology 110:53–72
    [Google Scholar]
  20. Hijmans E. M., Voorhoeve P. M., Beijersbergen R. L., Van ’T Veer L. J., Bernards R. 1995; E2F-5, a new E2F family member that interacts with p130 in vivo. Molecular and Cellular Biology 15:3082–3089
    [Google Scholar]
  21. Ho S. N., Hunt H. D., Horton R. M., Pullen J. K., Pease L. R. 1989; Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77:51–59
    [Google Scholar]
  22. Hoeffler W. K., Kovelmann R., Roeder R. G. 1988; Activation of transcription factor IIIC by the E1A protein. Cell 53:907–920
    [Google Scholar]
  23. Jones N. C. 1992; The multifunctional products of adenovirus E1A gene. In Malignant Transformation by DNA Viruses pp. 91–104 Doerfler W., Böhm P. Edited by New York: VHC Publishers;
    [Google Scholar]
  24. Kawamura H., Wada N., Makino Y., Tamura T. -A., Koikeda S., Shiroki K., Masamune Y., Nakanishi Y. 1994; Transcription stimulation of the adenovirus type 12 E1A gene in vitro by the 266-amino-acid E1A protein. Journal of Virology 68:5056–5062
    [Google Scholar]
  25. Kirch H. C., Pützer B., Schwabe G., Krüger H., Gnauck A., Schulte Holthausen H. 1993; Regulat ion of adenovirus 12 E1A transcription: E2F and ATF motifs in the E1A promoter bind nuclear protein complexes including E2F1, DP-1, cyclin A and/or RB and mediate transcriptional (auto)activation. Cellular and Molecular Biology Research 39:705–716
    [Google Scholar]
  26. Koikeda S., Ibuki R., Sawada Y., Nagata K., Shibata H., Masamue Y., Nakanishi Y. 1990; Nuclear factor I stimulates transcription of the adenovirus 12 E1A gene in a cell-free system. Biochimica et BiophysicaActa 1048:85–92
    [Google Scholar]
  27. Larsen P. L., Tibbetts C. 1987; Adenovirus gene autorepression: revertants of an E1A promoter mutation encode altered E1A proteins. Proceedings of the National Academy of Sciences, USA 84:8185–8189
    [Google Scholar]
  28. La Thangue N. B., Thimmapaya B., Rigby T. 1990; The embryonal carcinoma stem cell E1A-like activity involves a differentiation regulated transcription factor. Nucleic Acids Research 18:2929–2938
    [Google Scholar]
  29. Lavery D., Fu S. M., Lufkin T., Chen-Kiang S. 1987; Productive infection of cultured human lymphoid cells by adenovirus. Journal of Virology 61:1466–1472
    [Google Scholar]
  30. Lewin B. 1990; Commitment and activation at pol II promoters: a tail of protein-protein interactions. Cell 61:1161–1164
    [Google Scholar]
  31. Lynch M. J., Trainer D. L. 1989; Immortalization of primary baby rat kidney cells by retroviral mediated gene transfer of E1A genes. Cancer Research 49:5429–5434
    [Google Scholar]
  32. Manohar C. F., Kratochvil J., Thimmapaya B. 1990; The adenovirus EII early promoter has multiple E1A-sensitive elements, two of which function cooperatively in basal and virus-induced transcription. Journal of Virology 64:2457–2466
    [Google Scholar]
  33. Murre C., McCaw P. S., Vassin H., Claudy M., Jan L. Y., Jan Y. N., Cabrera C. V., Buskin J. N., Hauschka S. D., Lassar A. B., Weintraub H., Baltimore D. 1989; Interactions between heterologous helix- loop-helix proteins generate complexes that bind specifically to a common DNA sequence. Cell 58:537–544
    [Google Scholar]
  34. Nakanishi Y., Shibata H., Hase T., Masamune Y. 1987; Analysis of promoters of adenovirus type 12 E1A gene in a cell-free transcription system. Biochemical and Biophysical Research Communications 146:783–790
    [Google Scholar]
  35. Nakanishi Y., Koikeda S., Shibata H., Masamune Y. 1989; Interactions of factors bound at two different sites in the 5′-upstream region of the adenovirus 12 E1A gene. Biochemical and Biophysical Research Communications 158:685–689
    [Google Scholar]
  36. Neumann J. R., Morency C. A., Russian K. O. 1987; A novel rapid assay for chloramphenicol acetyltransferase gene expression. Bio/ Techniques 5:444–447
    [Google Scholar]
  37. Ou S. -H., Garcia-Martinez L. F., Paulssen E. J., Gaynor R. B. 1994; Role of flanking E box motifs in human immunodeficiency virus type 1 TATA element function. Journal of Virology 68:7188–7199
    [Google Scholar]
  38. Raychaudhuri P., Bagchi S., Nevins J. R. 1989; DNA-binding activity of the adenovirus-induced E4F transcription factor is regulated by phosphorylation. Genes & Development 3:620–627
    [Google Scholar]
  39. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  40. Saito I., Sato J. -I., Hanada H., Shiroki K., Shimojo H. 1981; Mapping of RNAs transcribed from adenovirus type 12 early and VA RNA regions. Virology 114:379–398
    [Google Scholar]
  41. Sawada Y., Fujinaga K. 1980; Mapping of adenovirus 12 mRNAs transcribed from the transforming region. Journal of Virology 36:639–651
    [Google Scholar]
  42. Shibata H., Zheng J. -H., Koikeda S., Masamune Y., Nakanishi Y. 1989; Cis- and trans- acting factors for transcription of the adenovirus 12 E1A gene. Biochimica et Biophysica Acta 1007:184–191
    [Google Scholar]
  43. Shibata-Sakurai H., Ando T., Masamune Y., Nakanishi Y. 1991; Transcription stimulation ofthe adenovirus type-12 E1A gene in vitro by a novel factor bound to a region adjacent to a TATA box. Gene 109:171–176
    [Google Scholar]
  44. Shibata-Sakurai H., Masamune Y., Nakanishi Y. 1993; Recognition of a palindromic DNA sequence by ESF-1, a factor stimulating transcription ofthe adenovirus type 12 E1A gene. Biological and Pharmaceutical Bulletin 16:87–89
    [Google Scholar]
  45. Shin E. K., Shin A., Paulding C., Schaffhausen B., Yee A. 1995; Multiple changes in E2F function and regulation occur upon muscle differentiation. Molecular and Cellular Biology 15:2252–2262
    [Google Scholar]
  46. Slansky J. E., Li Y., Kaelin W. G., Farnham P. J. 1993; A protein synthesis-dependent increase in E2F1 mRNA correlates with growth regulation of the dihydrofolate reductase promoter. Molecular and Cellular Biology 13:1610–1618
    [Google Scholar]
  47. Van den Hoff M. J. B., Moorman A. F. M., Lamers W. H. 1992; Electroporation in intracellular buffer increases cell survival. Nucleic Acids Research 20:2902
    [Google Scholar]
  48. Wolf D. A., Hermeking H., Albert T., Herzinger T., Kind P., Eick D. 1995; A complex between E2F and the Pb-related p130 is specifically targeted by the simian virus 40 large T antigen during cell transformation. Oncogene 10:2067–2078
    [Google Scholar]
  49. Yamazaki Y., Shimada Y., Shibata-Sakurai H., Masamune Y., Nakanishi Y. 1992; Multiple cis-acting DNA elements that regulate transcription of the adenovirus 12 E1A gene. Virus Genes 6:261–271
    [Google Scholar]
  50. Yee A. S., Raychaudhuri P., Jakoi L., Nevins J. R. 1989; The adenovirus-inducible factor E2F stimulates transcription after specific DNA binding. Molecular and Cellular Biology 9:578–585
    [Google Scholar]
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