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. 1982 Mar;79(6):1834–1838. doi: 10.1073/pnas.79.6.1834

Transcription of adenovirus cores in vitro: major RNA products differ from those made from a DNA template.

E Daniell, M J Fedor
PMCID: PMC346075  PMID: 6952232

Abstract

Intact cores of adenovirus type 5 were transcribed in vitro in extracts derived from uninfected HeLa cells, and the products were analyzed by hybridization to restricted viral DNA. Predominant transcripts from cores were distinctly different from transcripts generated from deproteinized DNA. The extracts contained RNA polymerases II and III. Transcription from cores was completely inhibited by low levels of alpha-amanitin, and there was little or no hybridization of products to the region of the genome encoding the small polymerase III-dependent virus-associated RNA that was a major transcript from the DNA template. The major transcripts from cores hybridized to the rightward-transcribed strand of the region between 75 and 89 map units, early region 3 of the adenovirus genome. There was several-fold more transcription of this region in cores than of the region downstream from the major late promoter, which predominates among polymerase II transcripts from DNA.

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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. 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]
  3. Brown D. T., Westphal M., Burlingham B. T., Winterhoff U., Doerfler W. Structure and composition of the adenovirus type 2 core. J Virol. 1975 Aug;16(2):366–387. doi: 10.1128/jvi.16.2.366-387.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Daniell E. Genome structure of incomplete particles of adenovirus. J Virol. 1976 Aug;19(2):685–708. doi: 10.1128/jvi.19.2.685-708.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daniell E., Groff D. E., Fedor M. J. Adenovirus chromatin structure at different stages of infection. Mol Cell Biol. 1981 Dec;1(12):1094–1105. doi: 10.1128/mcb.1.12.1094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Daniell E., Mullenbach T. Synthesis of defective viral DNA in HeLa cells infected with adenovirus type 3. J Virol. 1978 Apr;26(1):61–70. doi: 10.1128/jvi.26.1.61-70.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Engelke D. R., Ng S. Y., Shastry B. S., Roeder R. G. Specific interaction of a purified transcription factor with an internal control region of 5S RNA genes. Cell. 1980 Mar;19(3):717–728. doi: 10.1016/s0092-8674(80)80048-1. [DOI] [PubMed] [Google Scholar]
  9. Evans R., Weber J., Ziff E., Darnell J. E. Premature termination during adenovirus transcription. Nature. 1979 Mar 22;278(5702):367–370. doi: 10.1038/278367a0. [DOI] [PubMed] [Google Scholar]
  10. Everitt E., Lutter L., Philipson L. Structural proteins of adenoviruses. XII. Location and neighbor relationship among proteins of adenovirion type 2 as revealed by enzymatic iodination, immunoprecipitation and chemical cross-linking. Virology. 1975 Sep;67(1):197–208. doi: 10.1016/0042-6822(75)90417-1. [DOI] [PubMed] [Google Scholar]
  11. Flint S. J., Gallimore P. H., Sharp P. A. Comparison of viral RNA sequences in adenovirus 2-transformed and lytically infected cells. J Mol Biol. 1975 Jul 25;96(1):47–68. doi: 10.1016/0022-2836(75)90181-3. [DOI] [PubMed] [Google Scholar]
  12. Galos R. S., Williams J., Binger M. H., Flint S. J. Location of additional early gene sequences in the adenoviral chromosome. Cell. 1979 Aug;17(4):945–956. doi: 10.1016/0092-8674(79)90334-9. [DOI] [PubMed] [Google Scholar]
  13. Hammarskjöld M. L., Winberg G. Encapsidation of adenovirus 16 DNA is directed by a small DNA sequence at the left end of the genome. Cell. 1980 Jul;20(3):787–795. doi: 10.1016/0092-8674(80)90325-6. [DOI] [PubMed] [Google Scholar]
  14. Jacobsen H., Klenow H., Overgaard-Hansen K. The N-terminal amino-acid sequences of DNA polymerase I from Escherichia coli and of the large and the small fragments obtained by a limited proteolysis. Eur J Biochem. 1974 Jun 15;45(2):623–627. doi: 10.1111/j.1432-1033.1974.tb03588.x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Kitchingman G. R., Westphal H. The structure of adenovirus 2 early nuclear and cytoplasmic RNAs. J Mol Biol. 1980 Feb 15;137(1):23–48. doi: 10.1016/0022-2836(80)90155-2. [DOI] [PubMed] [Google Scholar]
  17. Lee D. C., Roeder R. G. Transcription of adenovirus type 2 genes in a cell-free system: apparent heterogeneity of initiation at some promoters. Mol Cell Biol. 1981 Jul;1(7):635–651. doi: 10.1128/mcb.1.7.635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lewis J. B., Mathews M. B. Control of adenovirus early gene expression: a class of immediate early products. Cell. 1980 Aug;21(1):303–313. doi: 10.1016/0092-8674(80)90138-5. [DOI] [PubMed] [Google Scholar]
  19. Manley J. L., Fire A., Cano A., Sharp P. A., Gefter M. L. DNA-dependent transcription of adenovirus genes in a soluble whole-cell extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3855–3859. doi: 10.1073/pnas.77.7.3855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mathews M. B. Binding of adenovirus VA RNA to mRNA: a possible role in splicing? Nature. 1980 Jun 19;285(5766):575–577. doi: 10.1038/285575a0. [DOI] [PubMed] [Google Scholar]
  21. Matsui T., Segall J., Weil P. A., Roeder R. G. Multiple factors required for accurate initiation of transcription by purified RNA polymerase II. J Biol Chem. 1980 Dec 25;255(24):11992–11996. [PubMed] [Google Scholar]
  22. Mirza M. A., Weber J. M. Structure of adenovirus chromatin as probed with restriction endonucleases. Virology. 1981 Jan 30;108(2):351–360. doi: 10.1016/0042-6822(81)90443-8. [DOI] [PubMed] [Google Scholar]
  23. Murray V., Holliday R. Mechanism for RNA splicing of gene transcripts. FEBS Lett. 1979 Oct 1;106(1):5–7. doi: 10.1016/0014-5793(79)80682-1. [DOI] [PubMed] [Google Scholar]
  24. Nevins J. R., Ginsberg H. S., Blanchard J. M., Wilson M. C., Darnell J. E., Jr Regulation of the primary expression of the early adenovirus transcription units. J Virol. 1979 Dec;32(3):727–733. doi: 10.1128/jvi.32.3.727-733.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Pelham H. R., Brown D. D. A specific transcription factor that can bind either the 5S RNA gene or 5S RNA. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4170–4174. doi: 10.1073/pnas.77.7.4170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Persson H., Monstein H. J., Akusjärvi G., Philipson L. Adenovirus early gene products may control viral mRNA accumulation and translation in vivo. Cell. 1981 Feb;23(2):485–496. doi: 10.1016/0092-8674(81)90144-6. [DOI] [PubMed] [Google Scholar]
  28. Prage L., Pettersson U., Höglund S., Lonberg-Holm K., Philipson L. Structural proteins of adenoviruses. IV. Sequential degradation of the adenovirus type 2 virion. Virology. 1970 Oct;42(2):341–358. doi: 10.1016/0042-6822(70)90278-3. [DOI] [PubMed] [Google Scholar]
  29. Prage L., Pettersson U. Structural proteins of adenoviruses. VII. Purification and properties of an arginine-rich core protein from adenovirus type 2 and type 3. Virology. 1971 Aug;45(2):364–373. doi: 10.1016/0042-6822(71)90337-0. [DOI] [PubMed] [Google Scholar]
  30. Sergeant A., Tigges M. A., Raskas H. J. Nucleosome-like structural subunits of intranuclear parental adenovirus type 2 DNA. J Virol. 1979 Mar;29(3):888–898. doi: 10.1128/jvi.29.3.888-898.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shaw A. R., Ziff E. B. Transcripts from the adenovirus-2 major late promoter yield a single early family of 3' coterminal mRNAs and five late families. Cell. 1980 Dec;22(3):905–916. doi: 10.1016/0092-8674(80)90568-1. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Sung M. T., Lischwe M. A., Richards J. C., Hosokawa K. Adenovirus chromatin I. Isolation and characterization of the major core protein VII and precursor Pro-VII. J Biol Chem. 1977 Jul 25;252(14):4981–4987. [PubMed] [Google Scholar]
  34. Tate V. E., Philipson L. Parental adenovirus DNA accumulates in nucleosome-like structures in infected cells. Nucleic Acids Res. 1979 Jun 25;6(8):2769–2785. doi: 10.1093/nar/6.8.2769. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Thomas G. P., Mathews M. B. DNA replication and the early to late transition in adenovirus infection. Cell. 1980 Nov;22(2 Pt 2):523–533. doi: 10.1016/0092-8674(80)90362-1. [DOI] [PubMed] [Google Scholar]
  36. Weil P. A., Luse D. S., Segall J., Roeder R. G. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979 Oct;18(2):469–484. doi: 10.1016/0092-8674(79)90065-5. [DOI] [PubMed] [Google Scholar]
  37. Weingärtner B., Keller W. Transcription and processing of adenoviral RNA by extracts from HeLa cells. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4092–4096. doi: 10.1073/pnas.78.7.4092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ziff E. B. Transcription and RNA processing by the DNA tumour viruses. Nature. 1980 Oct 9;287(5782):491–499. doi: 10.1038/287491a0. [DOI] [PubMed] [Google Scholar]

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