Skip to main content
The EMBO Journal logoLink to The EMBO Journal
. 1990 May;9(5):1615–1623. doi: 10.1002/j.1460-2075.1990.tb08281.x

A mitigator sequence in the downstream region of the major late promoter of adenovirus type 12 DNA.

C Zock 1, W Doerfler 1
PMCID: PMC551857  PMID: 2158446

Abstract

Human adenovirus type 12 (Ad12) replicates in permissive human host cells, but undergoes an abortive infection cycle in non-permissive hamster cells. Ad12 DNA cannot replicate and late viral genes are not expressed in hamster cells, whereas most of the early viral mRNAs are synthesized. We have shown previously that the major late promoter of Ad12 DNA (Ad12 MLP; nucleotides -228 to +435 relative to nucleotide +1 as the site of transcriptional initiation) does not function in uninfected or in Ad12-infected hamster BHK21 cells. The transcriptional defect of Ad12 DNA in hamster cells has thus been, at least partly, localized to the viral MLP. As expected, this construct is active in permissive human cells. Here, we show that the sequence between nucleotides +249 and +435 in the Ad12 MLP is in some way responsible for the late transcriptional block of this promoter in hamster cells. An Ad12 MLP--CAT construct comprising nucleotides -228 to +248 shows striking activity in hamster cells, and its activity is very markedly enhanced in Ad2- or Ad12-infected hamster or human cells compared with the nucleotide -228 to +435 construct. By using exonuclease Bal31, a series of Ad12 MLP--CAT gene assemblies were constructed which carry deletions of increasing lengths in the downstream part of the Ad12 MLP. Activity measurements of these constructs in BHK21 and in HeLa cells have located the presumptive mitigator element to the Ad12 sequence between nucleotides +320 and +352 of the MLP. It is also demonstrated that in the nucleotide -228 to +248 MLP construct, transcription is initiated at the authentic Ad12 MLP cap site after the transfection of both hamster and human cells. The localization of this cap site in the nucleotide sequence of the Ad12 MLP indicates the similarity to the comparable start site in the MLP of Ad2 DNA.(ABSTRACT TRUNCATED AT 250 WORDS)

Full text

PDF
1615

Images in this article

Selected References

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

  1. Akusjärvi G., Persson H. Controls of RNA splicing and termination in the major late adenovirus transcription unit. Nature. 1981 Jul 30;292(5822):420–426. doi: 10.1038/292420a0. [DOI] [PubMed] [Google Scholar]
  2. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. BABLANIAN R., EGGERS H. J., TAMM I. STUDIES ON THE MECHANISM OF POLIOVIRUS-INDUCED CELL DAMAGE. I. THE RELATION BETWEEN POLIOVIRUS,-INDUCED METABOLIC AND MORPHOLOGICAL ALTERATIONS IN CULTURED CELLS. Virology. 1965 May;26:100–113. doi: 10.1016/0042-6822(65)90030-9. [DOI] [PubMed] [Google Scholar]
  4. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  5. Carthew R. W., Chodosh L. A., Sharp P. A. An RNA polymerase II transcription factor binds to an upstream element in the adenovirus major late promoter. Cell. 1985 Dec;43(2 Pt 1):439–448. doi: 10.1016/0092-8674(85)90174-6. [DOI] [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. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dignam J. D., Lebovitz R. M., Roeder R. G. Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. Nucleic Acids Res. 1983 Mar 11;11(5):1475–1489. doi: 10.1093/nar/11.5.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Doerfler W. Integration of the deoxyribonucleic acid of adenovirus type 12 into the deoxyribonucleic acid of baby hamster kidney cells. J Virol. 1970 Nov;6(5):652–666. doi: 10.1128/jvi.6.5.652-666.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Doerfler W., Lundholm U., Hirsch-Kauffmann M. Intracellular forms of adenovirus deoxyribonucleic acid. I. Evidence for a deoxyribonucleic acid-protein complex in baby hamster kidney cells infected with adenovirus type 12. J Virol. 1972 Feb;9(2):297–308. doi: 10.1128/jvi.9.2.297-308.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Doerfler W. Nonproductive infection of baby hamster kidney cells (BHK21) with adenovirus type 12. Virology. 1969 Aug;38(4):587–606. doi: 10.1016/0042-6822(69)90179-2. [DOI] [PubMed] [Google Scholar]
  12. Doerfler W. The fate of the DNA of adenovirus type 12 in baby hamster kidney cells. Proc Natl Acad Sci U S A. 1968 Jun;60(2):636–643. doi: 10.1073/pnas.60.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Esche H., Schilling R., Doerfler W. In vitro translation of adenovirus type 12-specific mRNA isolated from infected and transformed cells. J Virol. 1979 Apr;30(1):21–31. doi: 10.1128/jvi.30.1.21-31.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Evans R. M., Fraser N., Ziff E., Weber J., Wilson M., Darnell J. E. The initiation sites for RNA transcription in Ad2 DNA. Cell. 1977 Nov;12(3):733–739. doi: 10.1016/0092-8674(77)90273-2. [DOI] [PubMed] [Google Scholar]
  15. Fanning E., Doerfler W. Intracellular forms of adenovirus DNA. V. Viral DNA sequences in hamster cells abortively infected and transformed with human adenovirus type 12. J Virol. 1976 Nov;20(2):373–383. doi: 10.1128/jvi.20.2.373-383.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fried M., Crothers D. M. Equilibria and kinetics of lac repressor-operator interactions by polyacrylamide gel electrophoresis. Nucleic Acids Res. 1981 Dec 11;9(23):6505–6525. doi: 10.1093/nar/9.23.6505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Garner M. M., Revzin A. A gel electrophoresis method for quantifying the binding of proteins to specific DNA regions: application to components of the Escherichia coli lactose operon regulatory system. Nucleic Acids Res. 1981 Jul 10;9(13):3047–3060. doi: 10.1093/nar/9.13.3047. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Graham F. L., van der Eb A. J. Transformation of rat cells by DNA of human adenovirus 5. Virology. 1973 Aug;54(2):536–539. doi: 10.1016/0042-6822(73)90163-3. [DOI] [PubMed] [Google Scholar]
  20. Grob U., Stüber K. GENEXPERT, a program system for nucleic acid sequence structural interpretation. Comput Appl Biosci. 1987 Sep;3(3):243–244. doi: 10.1093/bioinformatics/3.3.243. [DOI] [PubMed] [Google Scholar]
  21. Hasson T. B., Soloway P. D., Ornelles D. A., Doerfler W., Shenk T. Adenovirus L1 52- and 55-kilodalton proteins are required for assembly of virions. J Virol. 1989 Sep;63(9):3612–3621. doi: 10.1128/jvi.63.9.3612-3621.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jüttermann R., Weyer U., Doerfler W. Defect of adenovirus type 12 replication in hamster cells: absence of transcription of viral virus-associated and L1 RNAs. J Virol. 1989 Aug;63(8):3535–3540. doi: 10.1128/jvi.63.8.3535-3540.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Klenow H., Overgaard-Hansen K., Patkar S. A. Proteolytic cleavage fo native DNA polymerase into two different catalytic fragments. Influence of assay condtions on the change of exonuclease activity and polymerase activity accompanying cleavage. Eur J Biochem. 1971 Oct 14;22(3):371–381. doi: 10.1111/j.1432-1033.1971.tb01554.x. [DOI] [PubMed] [Google Scholar]
  24. Klimkait T., Doerfler W. Adenovirus types 2 and 5 functions elicit replication and late expression of adenovirus type 12 DNA in hamster cells. J Virol. 1985 Aug;55(2):466–474. doi: 10.1128/jvi.55.2.466-474.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Klimkait T., Doerfler W. E1B functions of type C adenoviruses play a role in the complementation of blocked adenovirus type 12 DNA replication and late gene transcription in hamster cells. Virology. 1987 Nov;161(1):109–120. doi: 10.1016/0042-6822(87)90176-0. [DOI] [PubMed] [Google Scholar]
  26. Kruczek I., Doerfler W. Expression of the chloramphenicol acetyltransferase gene in mammalian cells under the control of adenovirus type 12 promoters: effect of promoter methylation on gene expression. Proc Natl Acad Sci U S A. 1983 Dec;80(24):7586–7590. doi: 10.1073/pnas.80.24.7586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kuhlmann I., Achten S., Rudolph R., Doerfler W. Tumor induction by human adenovirus type 12 in hamsters: loss of the viral genome from adenovirus type 12-induced tumor cells is compatible with tumor formation. EMBO J. 1982;1(1):79–86. doi: 10.1002/j.1460-2075.1982.tb01128.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Langner K. D., Weyer U., Doerfler W. Trans effect of the E1 region of adenoviruses on the expression of a prokaryotic gene in mammalian cells: resistance to 5' -CCGG- 3' methylation. Proc Natl Acad Sci U S A. 1986 Mar;83(6):1598–1602. doi: 10.1073/pnas.83.6.1598. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  30. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  31. Nevins J. R., Wilson M. C. Regulation of adenovirus-2 gene expression at the level of transcriptional termination and RNA processing. Nature. 1981 Mar 12;290(5802):113–118. doi: 10.1038/290113a0. [DOI] [PubMed] [Google Scholar]
  32. Ortin J., Doerfler W. Transcription of the genome of adenovirus type 12. I. Viral mRNA in abortively infected and transformed cells. J Virol. 1975 Jan;15(1):27–35. doi: 10.1128/jvi.15.1.27-35.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ortin J., Scheidtmann K. H., Greenberg R., Westphal M., Doerfler W. Transcription of the genome of adenovirus type 12. III. Maps of stable RNA from productively infected human cells and abortively infected and transformed hamster cells. J Virol. 1976 Nov;20(2):355–372. doi: 10.1128/jvi.20.2.355-372.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Raska K., Jr, Strohl W. A. The response of BHK21 cells to infection with type 12 adenovirus. VI. Synthesis of virus-specific RNA. Virology. 1972 Mar;47(3):734–742. doi: 10.1016/0042-6822(72)90563-6. [DOI] [PubMed] [Google Scholar]
  35. Richardson C. C. Phosphorylation of nucleic acid by an enzyme from T4 bacteriophage-infected Escherichia coli. Proc Natl Acad Sci U S A. 1965 Jul;54(1):158–165. doi: 10.1073/pnas.54.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. 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]
  37. Scott M. R., Westphal K. H., Rigby P. W. Activation of mouse genes in transformed cells. Cell. 1983 Sep;34(2):557–567. doi: 10.1016/0092-8674(83)90388-4. [DOI] [PubMed] [Google Scholar]
  38. Shu L. M., Hong J. S., Wei Y. F., Engler J. A. Nucleotide sequence of the genes encoded in early region 2b of human adenovirus type 12. Gene. 1986;46(2-3):187–195. doi: 10.1016/0378-1119(86)90403-8. [DOI] [PubMed] [Google Scholar]
  39. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  40. Strohl W. A., Rouse H. C., Schlesinger R. W. Properties of cells derived from adenovirus-induced hamster tumors by long-term in vitro cultivation. II. Nature of the restricted response to type 2 adenovirus. Virology. 1966 Apr;28(4):645–658. doi: 10.1016/0042-6822(66)90249-2. [DOI] [PubMed] [Google Scholar]
  41. TRENTIN J. J., YABE Y., TAYLOR G. The quest for human cancer viruses. Science. 1962 Sep 14;137(3533):835–841. doi: 10.1126/science.137.3533.835. [DOI] [PubMed] [Google Scholar]
  42. Weisshaar B., Langner K. D., Jüttermann R., Müller U., Zock C., Klimkait T., Doerfler W. Reactivation of the methylation-inactivated late E2A promoter of adenovirus type 2 by E1A (13 S) functions. J Mol Biol. 1988 Jul 20;202(2):255–270. doi: 10.1016/0022-2836(88)90456-1. [DOI] [PubMed] [Google Scholar]
  43. Weyer U., Doerfler W. Species dependence of the major late promoter in adenovirus type 12 DNA. EMBO J. 1985 Nov;4(11):3015–3019. doi: 10.1002/j.1460-2075.1985.tb04037.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ziff E. B., Evans R. M. Coincidence of the promoter and capped 5' terminus of RNA from the adenovirus 2 major late transcription unit. Cell. 1978 Dec;15(4):1463–1475. doi: 10.1016/0092-8674(78)90070-3. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES