Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1975 Sep;16(3):662–673. doi: 10.1128/jvi.16.3.662-673.1975

Adenovirus transcription. II. RNA sequences complementary to simian virus 40 and adenovirus 2DNA in AD2+ND1- and AD2+ND3-infected cells.

S J Flint, Y Wewerka-Lutz, A S Levine, J Sambrook, P A Sharp
PMCID: PMC354714  PMID: 169392

Abstract

The genomes of the two nondefective adenovirus 2/simian virus 40 (Ad2/SV 40) hybrid viruses, nondefective Ad2/SV 40 hybrid virus 1 (Ad2+ND1) and nondefective hybrid virus 3 (Ad2+ND3), WERE FORMED BY A DELETION OF ABOUT 5% OF Ad2 DNA and insertion of part of the SV40 genome. We have compared the cytoplasmic RNA synthesized during both the early and late stages of lytic infection of human cells by these hybrid viruses to that expressed in Ad2-infected and SV40-infected cells. Separated strands of the six fragments of 32P-labeled Ad2 DNA produced by cleavage with the restriction endonuclease EcoRI (isolated from Escherichia coli) and the four fragments of 32P-labeled SV40 DNA produced by cleavage with both a restriction nuclease isolated from Haemophilus parainfluenzae, Hpa1, and EcoRI were prepared by electrophoresis of denatured DNA in agarose gels. The fraction of each fragment strand expressed as cytoplasmic RNA was determined by annealing fragmented 32P-labeled strands to an excess of cellular RNA extracted from infected cells. The segment of Ad2 DNA deleted from both hybrid virus genomes is transcribed into cytoplasmic mRNA during the early phase of Ad2 infection. Hence, we suggest that Ad2 codes for at least one "early" gene product which is nonessential for virus growth in cell culture. In both early Ad2+ND1 and Ad2+ND3-infected cells, 1,000 bases of Ad2 DNA adjacent to the integrated SV40 sequences are expressed as cytoplasmic RNA but are not similarly expressed in early Ad2-infected cells. The 3' termini of this early hybrid virus RNA maps in the vicinity of 0.18 on the conventional SV40 map and probably terminates at the same position as early lytic SV40 cytoplasmic RNA. Therefore, the base sequence in this region of SV40 DNA specifies the 3' termini of early messenger RNA present in both hybrid virus and SV40-infected cells.

Full text

PDF
662

Images in this article

666Image
on p.666

Selected References

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

  1. Botchan M., Ozanne B., Sugden B., Sharp P. A., Sambrook J. Viral DNA in transformed cells. III. The amounts of different regions of the SV40 genome present in a line of transformed mouse cells. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4183–4187. doi: 10.1073/pnas.71.10.4183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Danna K. J., Sack G. H., Jr, Nathans D. Studies of simian virus 40 DNA. VII. A cleavage map of the SV40 genome. J Mol Biol. 1973 Aug 5;78(2):363–376. doi: 10.1016/0022-2836(73)90122-8. [DOI] [PubMed] [Google Scholar]
  3. Dhar R., Weissman S. M., Zain B. S., Pan J., Lewis A. M., Jr The nucleotide sequence preceding an RNA polymerase initiation site on SV40 DNA. Part 2. The sequence of the early strand transcript. Nucleic Acids Res. 1974 Apr;1(4):595–611. doi: 10.1093/nar/1.4.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Grodzicker T., Anderson C., Sharp P. A., Sambrook J. Conditional lethal mutants of adenovirus 2-simian virus 40 hybrids. I. Host range mutants of Ad2+ND1. J Virol. 1974 Jun;13(6):1237–1244. doi: 10.1128/jvi.13.6.1237-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Hayward G. S. Gel electrophoretic separation of the complementary strands of bacteriophage DNA. Virology. 1972 Jul;49(1):342–344. doi: 10.1016/s0042-6822(72)80042-4. [DOI] [PubMed] [Google Scholar]
  6. Henry P. H., Schnipper L. E., Samaha R. J., Crumpacker C. S., Lewis A. M., Jr, Levine A. S. Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. VI. Characterization of the DNA from five nondefective hybrid viruses. J Virol. 1973 May;11(5):665–671. doi: 10.1128/jvi.11.5.665-671.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  8. Kelly T. J., Jr, Lewis A. M., Jr Use of nondefective adenovirus-simian virus 40 hybrids for mapping the simian virus 40 genome. J Virol. 1973 Sep;12(3):643–652. doi: 10.1128/jvi.12.3.643-652.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Khoury G., Byrne J. C., Martin M. A. Patterns of Simian Virus 40 DNA transcription after acute infection of permissive and nonpermissive cells. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1925–1928. doi: 10.1073/pnas.69.7.1925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Khoury G., Howley P., Nathans D., Martin M. Posttranscriptional selection of simian virus 40-specific RNA. J Virol. 1975 Feb;15(2):433–437. doi: 10.1128/jvi.15.2.433-437.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Khoury G., Lewis A. M., Jr, Oxman M. N., Levine A. S. Strand orientation of SV40 transcription in cells infected by non-defective adenovirus 2-SV40 hybrid viruses. Nat New Biol. 1973 Dec 19;246(155):202–205. doi: 10.1038/newbio246202a0. [DOI] [PubMed] [Google Scholar]
  12. Khoury G., Martin M. A., Lee T. N., Danna K. J., Nathans D. A map of simian virus 40 transcription sites expressed in productively infected cells. J Mol Biol. 1973 Aug 5;78(2):377–389. doi: 10.1016/0022-2836(73)90123-x. [DOI] [PubMed] [Google Scholar]
  13. Kumar A., Lindberg U. Characterization of messenger ribonucleoprotein and messenger RNA from KB cells. Proc Natl Acad Sci U S A. 1972 Mar;69(3):681–685. doi: 10.1073/pnas.69.3.681. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lewis A. M., Jr, Levin M. J., Wiese W. H., Crumpacker C. S., Henry P. H. A nondefective (competent) adenovirus-SV40 hybrid isolated from the AD.2-SV40 hybrid population. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1128–1135. doi: 10.1073/pnas.63.4.1128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lewis A. M., Jr, Levine A. S., Crumpacker C. S., Levin M. J., Samaha R. J., Henry P. H. Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. V. Isolation of additional hybrids which differ in their simian virus 40-specific biological properties. J Virol. 1973 May;11(5):655–664. doi: 10.1128/jvi.11.5.655-664.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lewis A. M., Jr, Rowe W. P. Studies on nondefective adenovirus-simian virus 40 hybrid viruses. I. A newly characterized simian virus 40 antigen induced by the Ad2+ND 1 virus. J Virol. 1971 Feb;7(2):189–197. doi: 10.1128/jvi.7.2.189-197.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Morrow J. F., Berg P. Cleavage of Simian virus 40 DNA at a unique site by a bacterial restriction enzyme. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3365–3369. doi: 10.1073/pnas.69.11.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Morrow J. F., Berg P., Kelly T. J., Jr, Lewis A. M., Jr Mapping of simian virus 40 early functions on the viral chromosome. J Virol. 1973 Sep;12(3):653–658. doi: 10.1128/jvi.12.3.653-658.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mulder C., Delius H. Specificity of the break produced by restricting endonuclease R 1 in Simian virus 40 DNA, as revealed by partial denaturation mapping. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3215–3219. doi: 10.1073/pnas.69.11.3215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mulder C., Sharp P. A., Delius H., Pettersson U. Specific fragmentation of DNA of adenovirus serotypes 3, 5, 7, and 12, and adeno-simian virus 40 hybrid virus Ad2+ND1 by restriction endonuclease R.EcoRI. J Virol. 1974 Jul;14(1):68–77. doi: 10.1128/jvi.14.1.68-77.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Oxman M. N., Levine A. S., Crumpacker C. S., Levin M. J., Henry P. H., Lewis A. M., Jr Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. IV. Characterization of the simian virus 40 ribonucleic acid species induced by wild-type simian virus 40 and by the nondefective hybrid virus, Ad2 + ND 1 . J Virol. 1971 Aug;8(2):215–224. doi: 10.1128/jvi.8.2.215-224.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Patch C. T., Lewis A. M., Jr, Levine A. S. Evidence for a transcription-control region of Simian virus 40 in the adenovirus 2--Simian virus 40 hybrid, Ad2+ND 1 . Proc Natl Acad Sci U S A. 1972 Nov;69(11):3375–3379. doi: 10.1073/pnas.69.11.3375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pettersson U., Mulder C., Deluis H., Sharp P. A. Cleavage of adenovirus type 2 DNA into six unique fragments by endonuclease R-RI. Proc Natl Acad Sci U S A. 1973 Jan;70(1):200–204. doi: 10.1073/pnas.70.1.200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pettersson U., Philipson L., Höglund S. Structural proteins of adenoviruses. I. Purification and characterization of the adenovirus type 2 hexon antigen. Virology. 1967 Dec;33(4):575–590. doi: 10.1016/0042-6822(67)90057-8. [DOI] [PubMed] [Google Scholar]
  25. Pettersson U., Philipson L. Synthesis of complementary RNA sequences during productive adenovirus infection. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4887–4891. doi: 10.1073/pnas.71.12.4887. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. RABSON A. S., O'CONOR G. T., BEREZESKY I. K., PAUL F. J. ENHANCEMENT OF ADENOVIRUS GROWTH IN AFRICAN GREEN MONKEY KIDNEY CELL CULTURES BY SV40. Proc Soc Exp Biol Med. 1964 May;116:187–190. doi: 10.3181/00379727-116-29197. [DOI] [PubMed] [Google Scholar]
  27. Sambrook J., Sharp P. A., Keller W. Transcription of Simian virus 40. I. Separation of the strands of SV40 DNA and hybridization of the separated strands to RNA extracted from lytically infected and transformed cells. J Mol Biol. 1972 Sep 14;70(1):57–71. doi: 10.1016/0022-2836(72)90163-5. [DOI] [PubMed] [Google Scholar]
  28. Sambrook J., Sugden B., Keller W., Sharp P. A. Transcription of simian virus 40. 3. Mapping of "early" and "late" species of RNA. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3711–3715. doi: 10.1073/pnas.70.12.3711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. 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]
  30. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  31. Tal J., Craig E. A., Zimmer S., Raskas H. J. Localization of adenovirus 2 messenger RNA's to segments of the viral genome defined by endonuclease R-R1. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4057–4061. doi: 10.1073/pnas.71.10.4057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tibbetts C., Pettersson U. Complementary strand-specific sequences from unique fragments of adenovirus type 2 DNA for hybridization-mapping experiments. J Mol Biol. 1974 Oct 5;88(4):767–784. doi: 10.1016/0022-2836(74)90398-2. [DOI] [PubMed] [Google Scholar]
  33. Westphal H. SV40 DNA strand selection by Escherichia coli RNA polymerase. J Mol Biol. 1970 Jun 14;50(2):407–420. doi: 10.1016/0022-2836(70)90201-9. [DOI] [PubMed] [Google Scholar]
  34. Williams J. F. Enhancement of adenovirus plaque formation on HeLa cells by magnesium chloride. J Gen Virol. 1970 Dec;9(3):251–255. doi: 10.1099/0022-1317-9-3-251. [DOI] [PubMed] [Google Scholar]
  35. Wilson D. A., Thomas C. A., Jr Hydroxyapatite chromatography of short double-helical DNA. Biochim Biophys Acta. 1973 Dec 21;331(3):333–340. doi: 10.1016/0005-2787(73)90019-1. [DOI] [PubMed] [Google Scholar]

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

RESOURCES