Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1979 Dec;32(3):727–733. doi: 10.1128/jvi.32.3.727-733.1979

Regulation of the Primary Expression of the Early Adenovirus Transcription Units

Joseph R Nevins 1, Harold S Ginsberg 1,†, J-M Blanchard 1,‡, Michael C Wilson 1, James E Darnell Jr 1
PMCID: PMC525919  PMID: 513202

Abstract

The time course of appearance of transcriptional activity from five early adenovirus type 2 transcription units has been determined. RNA complementary to region 1A (1-4.4 map units), the first region to be transcribed, was detectable at 45 min after infection; a maximal rate of RNA synthesis was reached at 3 h after infection and was maintained thereafter for at least 6 h. RNA from region 2 (75-56 map units), which encodes the mRNA for the 72,000-dalton DNA-binding protein, was the last to be synthesized; transcription commenced at about 2 h postinfection, reached a maximum at 7 h, and then declined. Transcription of regions 3 (76-86 map units) and 4 (99-91 map units) reached a maximal value at 3 h postinfection. The rates of RNA synthesis from these regions then declined over the next 6 h. The decline of transcription from regions 2 and 4 appeared to be a specific repression of these transcription units. The repression did not occur in the absence of protein synthesis, suggesting that a viral protein might be involved. Transcription of all early regions was initiated and continued for at least 2 to 3 h in cells that were treated with cycloheximide or emetine before and during infection, suggesting that at least the initiation of RNA synthesis from the five early adenovirus type 2 transcription units does not depend on the formation of a viral protein. Moreover, mRNA was formed in the absence of protein synthesis that hybridized to DNA fragments representing each of the five early transcription units. The increase in mRNA accumulation in the presence of cycloheximide (or emetine) does not appear to be due to increased RNA synthesis; thus, either increased mRNA stability or increased efficiency of nuclear RNA processing must occur.

Full text

PDF
727

Selected References

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

  1. 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]
  2. Berk A. J., Sharp P. A. Ultraviolet mapping of the adenovirus 2 early promoters. Cell. 1977 Sep;12(1):45–55. doi: 10.1016/0092-8674(77)90184-2. [DOI] [PubMed] [Google Scholar]
  3. Blanchard J. M., Weber J., Jelinek W., Darnell J. E. In vitro RNA-RNA splicing in adenovirus 2 mRNA formation. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5344–5348. doi: 10.1073/pnas.75.11.5344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blanton R. A., Carter T. H. Autoregulation of adenovirus type 5 early gene expression. III. Transcription studies in isolated nuclei. J Virol. 1979 Feb;29(2):458–465. doi: 10.1128/jvi.29.2.458-465.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carter T. H., Blanton R. A. Autoregulation of adenovirus type 5 early gene expression II. Effect of temperature-sensitive early mutations on virus RNA accumulation. J Virol. 1978 Nov;28(2):450–456. doi: 10.1128/jvi.28.2.450-456.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Carter T. H., Blanton R. A. Possible role of the 72,000 dalton DNA-binding protein in regulation of adenovirus type 5 early gene expression. J Virol. 1978 Feb;25(2):664–674. doi: 10.1128/jvi.25.2.664-674.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chow L. T., Roberts J. M., Lewis J. B., Broker T. R. A map of cytoplasmic RNA transcripts from lytic adenovirus type 2, determined by electron microscopy of RNA:DNA hybrids. Cell. 1977 Aug;11(4):819–836. doi: 10.1016/0092-8674(77)90294-x. [DOI] [PubMed] [Google Scholar]
  8. Craig E. A., Raskas H. J. Effect of cycloheximide on RNA metabolism early in productive infection with adenovirus 2. J Virol. 1974 Jul;14(1):26–32. doi: 10.1128/jvi.14.1.26-32.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Eggerding F., Raskas H. J. Effect of protein synthesis inhibitors on viral mRNA's synthesized early in adenovirus type 2 infection. J Virol. 1978 Jan;25(1):453–458. doi: 10.1128/jvi.25.1.453-458.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eggerding F., Raskas H. J. Regulation of an early RNA transcribed from the transforming segment of the adenovirus 2 genome. Virology. 1978 Dec;91(2):312–320. doi: 10.1016/0042-6822(78)90379-3. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Ginsberg H. S., Ensinger M. J., Kauffman R. S., Mayer A. J., Lundholm U. Cell transformation: a study of regulation with types 5 and 12 adenovirus temperature-sensitive mutants. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):419–426. doi: 10.1101/sqb.1974.039.01.054. [DOI] [PubMed] [Google Scholar]
  13. Goldenberg C. J., Raskas H. J. Splicing patterns of nuclear precursors to the mRNA for adenovirus 2 DNA binding protein. Cell. 1979 Jan;16(1):131–138. doi: 10.1016/0092-8674(79)90194-6. [DOI] [PubMed] [Google Scholar]
  14. Grollman A. P. Inhibitors of protein biosynthesis. V. Effects of emetine on protein and nucleic acid biosynthesis in HeLa cells. J Biol Chem. 1968 Aug 10;243(15):4089–4094. [PubMed] [Google Scholar]
  15. Jelinek W., Molloy G., Fernandez-Munoz R., Salditt M., Darnell J. E. Secondary structure in heterogeneous nuclear RNA: involvement of regions from repeated DNA sites. J Mol Biol. 1974 Jan 25;82(3):361–370. doi: 10.1016/0022-2836(74)90597-x. [DOI] [PubMed] [Google Scholar]
  16. Kitchingman G. R., Lai S. P., Westphal H. Loop structures in hybrids of early RNA and the separated strands of adenovirus DNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4392–4395. doi: 10.1073/pnas.74.10.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levine A. J., Van Der Vliet P. C., Rosenwirth B., Rabek J., Frenkel G., Ensinger M. Adenovirus-infected, cell-specific, DNA-binding proteins. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):559–566. doi: 10.1101/sqb.1974.039.01.069. [DOI] [PubMed] [Google Scholar]
  18. Neuwald P. D., Meyer J., Maizel J. V., Jr, Westpahl H. Early gene expression of adenovirus type 2: R-loop mapping of mRNA and time course of viral DNA, mRNA, and protein synthesis. J Virol. 1977 Mar;21(3):1019–1030. doi: 10.1128/jvi.21.3.1019-1030.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Nevins J. R., Darnell J. E. Groups of adenovirus type 2 mRNA's derived from a large primary transcript: probable nuclear origin and possible common 3' ends. J Virol. 1978 Mar;25(3):811–823. doi: 10.1128/jvi.25.3.811-823.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nevins J. R., Darnell J. E., Jr Steps in the processing of Ad2 mRNA: poly(A)+ nuclear sequences are conserved and poly(A) addition precedes splicing. Cell. 1978 Dec;15(4):1477–1493. doi: 10.1016/0092-8674(78)90071-5. [DOI] [PubMed] [Google Scholar]
  21. Nevins J. R. Definition and mapping of adenovirus 2 nuclear transcription. Methods Enzymol. 1980;65(1):768–785. doi: 10.1016/s0076-6879(80)65072-1. [DOI] [PubMed] [Google Scholar]
  22. Nevins J. R. Processing of late adenovirus nuclear RNA to mRNA. Kinetics of formation of intermediates and demonstration that all events are nuclear. J Mol Biol. 1979 Jun 5;130(4):493–506. doi: 10.1016/0022-2836(79)90436-4. [DOI] [PubMed] [Google Scholar]
  23. Parsons J. T., Green M. Biochemical studies on adenovirus multiplication. 18. Resolution of early virus-specific RNA species in Ad 2 infected and transformed cells. Virology. 1971 Jul;45(1):154–162. doi: 10.1016/0042-6822(71)90122-x. [DOI] [PubMed] [Google Scholar]
  24. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  25. Pettersson U., Tibbetts C., Philipson L. Hybridization maps of early and late messenger RNA sequences on the adenovirus type 2 genome. J Mol Biol. 1976 Mar 15;101(4):479–501. doi: 10.1016/0022-2836(76)90241-2. [DOI] [PubMed] [Google Scholar]
  26. Sehgal P. B., Fraser N. W., Darnell J. E., Jr Early Ad-2 transcription units: only promoter-proximal RNA continues to be made in the presence of DRB. Virology. 1979 Apr 15;94(1):185–191. doi: 10.1016/0042-6822(79)90448-3. [DOI] [PubMed] [Google Scholar]
  27. Sharp P. A., Gallimore P. H., Flint S. J. Mapping of adenovirus 2 RNA sequences in lytically infected cells and transformed cell lines. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):457–474. doi: 10.1101/sqb.1974.039.01.058. [DOI] [PubMed] [Google Scholar]
  28. Soeiro R., Darnell J. E. Competition hybridization by "pre-saturation" of HeLa cell DNA. J Mol Biol. 1969 Sep 28;44(3):551–562. doi: 10.1016/0022-2836(69)90379-9. [DOI] [PubMed] [Google Scholar]
  29. Spector D. J., McGrogan M., Raskas H. J. Regulation of the appearance of cytoplasmic RNAs from region 1 of the adenovirus 2 genome. J Mol Biol. 1978 Dec 15;126(3):395–414. doi: 10.1016/0022-2836(78)90048-7. [DOI] [PubMed] [Google Scholar]
  30. Van Der Vliet P. C., Levine A. J., Ensinger M. J., Ginsberg H. S. Thermolabile DNA binding proteins from cells infected with a temperature-sensitive mutant of adenovrius defective in viral DNA synthesis. J Virol. 1975 Feb;15(2):348–354. doi: 10.1128/jvi.15.2.348-354.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Vliet P. C., Sussenbach J. S. An adenovirus type 5 gene function required for initiation of viral DNA replication. Virology. 1975 Oct;67(2):415–426. doi: 10.1016/0042-6822(75)90443-2. [DOI] [PubMed] [Google Scholar]
  32. Wall R., Philipson L., Darnell J. E. Processing of adenovirus specific nuclear RNA during virus replication. Virology. 1972 Oct;50(1):27–34. doi: 10.1016/0042-6822(72)90342-x. [DOI] [PubMed] [Google Scholar]
  33. Wilson M. C., Fraser N. W., Darnell J. E., Jr Mapping of RNA initiation sites by high doses of uv irradiation: evidence for three independent promoters within the left 11% of the Ad-2 genome. Virology. 1979 Apr 15;94(1):175–184. doi: 10.1016/0042-6822(79)90447-1. [DOI] [PubMed] [Google Scholar]
  34. Wilson M. C., Sawicki S. G., Salditt-Georgieff M., Darnell J. E. Adenovirus type 2 mRNA in transformed cells: map positions and difference in transport time. J Virol. 1978 Jan;25(1):97–103. doi: 10.1128/jvi.25.1.97-103.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. 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]
  36. van der Vliet P. C., Levine A. J. DNA-binding proteins specific for cells infected by adenovirus. Nat New Biol. 1973 Dec 12;246(154):170–174. doi: 10.1038/newbio246170a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES