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. 1984 Oct 25;12(20):7877–7888. doi: 10.1093/nar/12.20.7877

The E4 transcriptional unit of Ad2: far upstream sequences are required for its transactivation by E1A.

P Gilardi, M Perricaudet
PMCID: PMC320206  PMID: 6093057

Abstract

We have investigated the effect of the E1A polypeptides of adenovirus 2 on the transcription of the viral E4 region. For this purpose, we have fused the promoter region of the E4 gene to the bacterial gene coding for chloramphenicol acetyl transferase. We have found that transcription from the E4 promoter is increased at least 20 fold in the presence of the E1A region. We have also found that the largest E1A polypeptide is the regulating factor, whereas the shortest has no apparent effect. Deletion of sequences upstream from position -158, as measured from the cap site, reduces the efficiency of the transcription in the presence of E1A by more than 15 fold. An important regulatory domain lies between positions -158 and -179. This domain contains the sequence 5' GGGAAGTGAC 3' which is homologous to the E1A enhancer core sequence. Another similar sequence is also present at position -149.

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Selected References

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  1. Aiello L., Guilfoyle R., Huebner K., Weinmann R. Adenovirus 5 DNA sequences present and RNA sequences transcribed in transformed human embryo kidney cells (HEK-Ad-5 or 293). Virology. 1979 Apr 30;94(2):460–469. doi: 10.1016/0042-6822(79)90476-8. [DOI] [PubMed] [Google Scholar]
  2. Baker C. C., Ziff E. B. Promoters and heterogeneous 5' termini of the messenger RNAs of adenovirus serotype 2. J Mol Biol. 1981 Jun 25;149(2):189–221. doi: 10.1016/0022-2836(81)90298-9. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Berk A. J., Sharp P. A. Structure of the adenovirus 2 early mRNAs. Cell. 1978 Jul;14(3):695–711. doi: 10.1016/0092-8674(78)90252-0. [DOI] [PubMed] [Google Scholar]
  5. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chow L. T., Broker T. R., Lewis J. B. Complex splicing patterns of RNAs from the early regions of adenovirus-2. J Mol Biol. 1979 Oct 25;134(2):265–303. doi: 10.1016/0022-2836(79)90036-6. [DOI] [PubMed] [Google Scholar]
  7. Elkaim R., Goding C., Kédinger C. The adenovirus-2 EIIa early gene promoter: sequences required for efficient in vitro and in vivo transcription. Nucleic Acids Res. 1983 Oct 25;11(20):7105–7117. doi: 10.1093/nar/11.20.7105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gaynor R. B., Berk A. J. Cis-acting induction of adenovirus transcription. Cell. 1983 Jul;33(3):683–693. doi: 10.1016/0092-8674(83)90011-9. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  11. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  12. Handa H., Sharp P. A. Requirement for distal upstream sequences for maximal transcription in vitro of early region IV of adenovirus. Mol Cell Biol. 1984 Apr;4(4):791–798. doi: 10.1128/mcb.4.4.791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hearing P., Shenk T. The adenovirus type 5 E1A transcriptional control region contains a duplicated enhancer element. Cell. 1983 Jul;33(3):695–703. doi: 10.1016/0092-8674(83)90012-0. [DOI] [PubMed] [Google Scholar]
  14. Hérissé J., Rigolet M., de Dinechin S. D., Galibert F. Nucleotide sequence of adenovirus 2 DNA fragment encoding for the carboxylic region of the fiber protein and the entire E4 region. Nucleic Acids Res. 1981 Aug 25;9(16):4023–4042. doi: 10.1093/nar/9.16.4023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Jones N., Shenk T. An adenovirus type 5 early gene function regulates expression of other early viral genes. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3665–3669. doi: 10.1073/pnas.76.8.3665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Leff T., Elkaim R., Goding C. R., Jalinot P., Sassone-Corsi P., Perricaudet M., Kédinger C., Chambon P. Individual products of the adenovirus 12S and 13S EIa mRNAs stimulate viral EIIa and EIII expression at the transcriptional level. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4381–4385. doi: 10.1073/pnas.81.14.4381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lusky M., Botchan M. Inhibition of SV40 replication in simian cells by specific pBR322 DNA sequences. Nature. 1981 Sep 3;293(5827):79–81. doi: 10.1038/293079a0. [DOI] [PubMed] [Google Scholar]
  19. Messing J., Vieira J. A new pair of M13 vectors for selecting either DNA strand of double-digest restriction fragments. Gene. 1982 Oct;19(3):269–276. doi: 10.1016/0378-1119(82)90016-6. [DOI] [PubMed] [Google Scholar]
  20. Montell C., Fisher E. F., Caruthers M. H., Berk A. J. Resolving the functions of overlapping viral genes by site-specific mutagenesis at a mRNA splice site. Nature. 1982 Feb 4;295(5848):380–384. doi: 10.1038/295380a0. [DOI] [PubMed] [Google Scholar]
  21. Nevins J. R. Mechanism of activation of early viral transcription by the adenovirus E1A gene product. Cell. 1981 Oct;26(2 Pt 2):213–220. doi: 10.1016/0092-8674(81)90304-4. [DOI] [PubMed] [Google Scholar]
  22. Perricaudet M., Akusjärvi G., Virtanen A., Pettersson U. Structure of two spliced mRNAs from the transforming region of human subgroup C adenoviruses. Nature. 1979 Oct 25;281(5733):694–696. doi: 10.1038/281694a0. [DOI] [PubMed] [Google Scholar]
  23. Prentki P., Krisch H. M. A modified pBR322 vector with improved properties for the cloning, recovery, and sequencing of blunt-ended DNA fragments. Gene. 1982 Feb;17(2):189–196. doi: 10.1016/0378-1119(82)90072-5. [DOI] [PubMed] [Google Scholar]
  24. Ricciardi R. P., Jones R. L., Cepko C. L., Sharp P. A., Roberts B. E. Expression of early adenovirus genes requires a viral encoded acidic polypeptide. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6121–6125. doi: 10.1073/pnas.78.10.6121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Richardson W. D., Westphal H. A cascade of adenovirus early functions is required for expression of adeno-associated virus. Cell. 1981 Nov;27(1 Pt 2):133–141. doi: 10.1016/0092-8674(81)90367-6. [DOI] [PubMed] [Google Scholar]
  26. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spector D. J. The pattern of integration of viral DNA sequences in the adenovirus 5-transformed human cell line 293. Virology. 1983 Oct 30;130(2):533–538. doi: 10.1016/0042-6822(83)90107-1. [DOI] [PubMed] [Google Scholar]
  28. Svensson C., Akusjärvi G. Adenovirus 2 early region 1A stimulates expression of both viral and cellular genes. EMBO J. 1984 Apr;3(4):789–794. doi: 10.1002/j.1460-2075.1984.tb01886.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Svensson C., Pettersson U., Akusjärvi G. Splicing of adenovirus 2 early region 1A mRNAs is non-sequential. J Mol Biol. 1983 Apr 15;165(3):475–495. doi: 10.1016/s0022-2836(83)80214-9. [DOI] [PubMed] [Google Scholar]
  30. Weeks D. L., Jones N. C. E1A control of gene expression is mediated by sequences 5' to the transcriptional starts of the early viral genes. Mol Cell Biol. 1983 Jul;3(7):1222–1234. doi: 10.1128/mcb.3.7.1222. [DOI] [PMC free article] [PubMed] [Google Scholar]

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