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. 1980 Oct;36(1):143–151. doi: 10.1128/jvi.36.1.143-151.1980

New chimeric splice junction in adenovirus type 2-simian virus 40 hybrid viral mRNA.

G Khoury, J Alwine, N Goldman, P Gruss, G Jay
PMCID: PMC353625  PMID: 6255201

Abstract

We have examined hybrid viral RNAs synthesized in both human and monkey cells infected by three nondefective adenovirus type 2 (Ad2)-simian virus 40 (SV40) hybrid viruses; Ad2+ND1, Ad2+ND2, and Ad2+ND4. Most of the hybrid viral RNA molecules appeared to be initiated within adenoviral sequences, but were polyadenylated on their 3' end at the early SV40 mRNA polyadenylation site. The Ad2+ND4 stock of virus was not homogeneous, but consisted of two principle populations of viral DNA. Both populations contained a segment of SV40 DNA extending from the SV40 map positions 0.63 to 0.11 in a left-to-right orientation at adenovirus map position 0.82. One population contained an intact SV40 segment, whereas the other (representing 80 to 85% of the population) has a 500-base pair deletion mapping from approximately 0.60 to 0.50 SV40 map units. This deletion encompassed the SV40 DNA segment which encodes the early SV40 splice sites. Cells infected by the mixed Ad2+ND4 population induced the synthesis of at least three major SV40 RNA species among the hybrid viral transcripts. The most abundant of these hybrid mRNA's appeared only late in the lytic cycle, after the onset of viral DNA replication. It contained an RNA splice junction which extended from a donor (5') nucleotide within the adenoviral RNA sequences to an acceptor (3') splice site within the early region of SV40 at 0.46 SV40 map units. This SV40 acceptor splice site was remarkable in that its use has not been detected in the spliced viral mRNA's of SV40-infected or -transformed cells.

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Selected References

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  1. Adams S. L., Alwine J. C., de Crombrugghe B., Pastan I. Use of recombinant plasmids to characterize collagen RNAs in normal and transformed chick embryo fibroblasts. J Biol Chem. 1979 Jun 25;254(12):4935–4938. [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. Alwine J. C., Khoury G. Control of simian virus 40 gene expression at the levels of RNA synthesis and processing: thermally induced changes in the ratio of the simian virus 40 early mRNA's and proteins. J Virol. 1980 Jul;35(1):157–164. doi: 10.1128/jvi.35.1.157-164.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bachenheimer S., Darnell J. E. Adenovirus-2 mRNA is transcribed as part of a high-molecular-weight precursor RNA. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4445–4449. doi: 10.1073/pnas.72.11.4445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  6. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Chow L. T., Broker T. R. The spliced structures of adenovirus 2 fiber message and the other late mRNAs. Cell. 1978 Oct;15(2):497–510. doi: 10.1016/0092-8674(78)90019-3. [DOI] [PubMed] [Google Scholar]
  10. Chow L. T., Broker T. R. The spliced structures of adenovirus 2 fiber message and the other late mRNAs. Cell. 1978 Oct;15(2):497–510. doi: 10.1016/0092-8674(78)90019-3. [DOI] [PubMed] [Google Scholar]
  11. Crick F. Split genes and RNA splicing. Science. 1979 Apr 20;204(4390):264–271. doi: 10.1126/science.373120. [DOI] [PubMed] [Google Scholar]
  12. Darnell J. E., Jr Implications of RNA-RNA splicing in evolution of eukaryotic cells. Science. 1978 Dec 22;202(4374):1257–1260. doi: 10.1126/science.364651. [DOI] [PubMed] [Google Scholar]
  13. Deppert W., Walter G. Simian virus to (SV40) tumor-specific proteins in nucleus and plasma membrane of HeLa cells infected by adenovirus 2-SV40 hybrid virus Ad2+ND2. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2505–2509. doi: 10.1073/pnas.73.7.2505. [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. Flint J. The topography and transcription of the adenovirus genome. Cell. 1977 Feb;10(2):153–166. doi: 10.1016/0092-8674(77)90211-2. [DOI] [PubMed] [Google Scholar]
  16. Flint S. J., Wewerka-Lutz Y., Levine A. S., Sambrook J., Sharp P. A. Adenovirus transcription. II. RNA sequences complementary to simian virus 40 and adenovirus 2DNA in AD2+ND1- and AD2+ND3-infected cells. J Virol. 1975 Sep;16(3):662–673. doi: 10.1128/jvi.16.3.662-673.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Goldberg S., Weber J., Darnell J. E., Jr The definition of a large viral transcription unit late in Ad2 infection of HeLa cells: mapping by effects of ultraviolet irradiation. Cell. 1977 Apr;10(4):617–621. doi: 10.1016/0092-8674(77)90094-0. [DOI] [PubMed] [Google Scholar]
  18. Gruss P., Lai C. J., Dhar R., Khoury G. Splicing as a requirement for biogenesis of functional 16S mRNA of simian virus 40. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4317–4321. doi: 10.1073/pnas.76.9.4317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Halbert D. N., Spector D. J., Raskas H. J. In vitro translation products specified by the transforming region of adenovirus type 2. J Virol. 1979 Sep;31(3):621–629. doi: 10.1128/jvi.31.3.621-629.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hamer D. H., Leder P. Splicing and the formation of stable RNA. Cell. 1979 Dec;18(4):1299–1302. doi: 10.1016/0092-8674(79)90240-x. [DOI] [PubMed] [Google Scholar]
  21. Jay G., Jay F. T., Friedman R. M., Levine A. S. Biosynthesis, immunological specificity, and intracellular distribution of the simian virus 40-specific protein induced by the nondefective hybrid Ad2+ND1. J Virol. 1978 May;26(2):411–419. doi: 10.1128/jvi.26.2.411-419.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jay G., Jay F. T., Friedman R. M., Levine A. S. Simian virus 40-specific ribosome-binding proteins induced by a nondefective adenovirus 2-simian virus 40 hybrid. J Virol. 1977 Sep;23(3):692–699. doi: 10.1128/jvi.23.3.692-699.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Khoury G., Gruss P., Dhar R., Lai C. J. Processing and expression of early SV40 mRNA: a role for RNA conformation in splicing. Cell. 1979 Sep;18(1):85–92. doi: 10.1016/0092-8674(79)90356-8. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Lai C. J., Khoury G. Deletion mutants of simian virus 40 defective in biosynthesis of late viral mRNA. Proc Natl Acad Sci U S A. 1979 Jan;76(1):71–75. doi: 10.1073/pnas.76.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lawrence C., Hunter T., Walter G. Characterization of the simian virus 40-specific messenger RNAs isolated from HeLa cells infected with the non-defective adenovirus 2-simian virus 40 hybrid viruses Ad2+ND2 and Ad2+ND4. J Mol Biol. 1979 Oct 15;134(1):95–108. doi: 10.1016/0022-2836(79)90415-7. [DOI] [PubMed] [Google Scholar]
  27. Lerner M. R., Boyle J. A., Mount S. M., Wolin S. L., Steitz J. A. Are snRNPs involved in splicing? Nature. 1980 Jan 10;283(5743):220–224. doi: 10.1038/283220a0. [DOI] [PubMed] [Google Scholar]
  28. Lewis A. M., Jr Defective and nondefective Ad2-SV40 hybrids. Prog Med Virol. 1977;23:96–139. [PubMed] [Google Scholar]
  29. 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]
  30. Mann K., Hunter T., Walter G., Linke H. Evidence for simian virus 40 (SV40) coding of SV40 T-antigen and the SV40-specific proteins in HeLa cells infected with nondefective adenovirus type 2-SV40 hybrid viruses. J Virol. 1977 Oct;24(1):151–169. doi: 10.1128/jvi.24.1.151-169.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Mulligan R. C., Howard B. H., Berg P. Synthesis of rabbit beta-globin in cultured monkey kidney cells following infection with a SV40 beta-globin recombinant genome. Nature. 1979 Jan 11;277(5692):108–114. doi: 10.1038/277108a0. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Oxman M. N., Levin M. J., Lewis A. M., Jr Control of simian virus 40 gene expression in adenovirus-simian virus 40 hybrid viruses. Synthesis of hybrid adenovirus 2-simian virus 40 RNA molecules in cells infected with a nondefective adenovirus 2-simian virus 40 hybrid virus. J Virol. 1974 Feb;13(2):322–330. doi: 10.1128/jvi.13.2.322-330.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Reddy V. B., Ghosh P. K., Lebowitz P., Piatak M., Weissman S. M. Simian virus 40 early mRNA's. I. Genomic localization of 3' and 5' termini and two major splices in mRNA from transformed and lytically infected cells. J Virol. 1979 Apr;30(1):279–296. doi: 10.1128/jvi.30.1.279-296.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  37. Rogers J., Wall R. A mechanism for RNA splicing. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1877–1879. doi: 10.1073/pnas.77.4.1877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weber J., Jelinek W., Darnell J. E., Jr The definition of a large viral transcription unit late in Ad2 infection of HeLa cells: mapping of nascent RNA molecules labeled in isolated nuclei. Cell. 1977 Apr;10(4):611–616. doi: 10.1016/0092-8674(77)90093-9. [DOI] [PubMed] [Google Scholar]
  39. Westphal H., Lai S. P., Lawrence C., Hunter T., Walter G. Mosaic adenovirus-SV40 RNA specified by the non-defective hybrid virus Ad2+ND4. J Mol Biol. 1979 May 25;130(3):337–351. doi: 10.1016/0022-2836(79)90545-x. [DOI] [PubMed] [Google Scholar]
  40. Westphal H., Lai S. P. Quantitative electron microscopy of early adenovirus RNA. J Mol Biol. 1977 Nov 5;116(3):525–548. doi: 10.1016/0022-2836(77)90082-1. [DOI] [PubMed] [Google Scholar]
  41. Zain S., Sambrook J., Roberts R. J., Keller W., Fried M., Dunn A. R. Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA. Cell. 1979 Apr;16(4):851–861. doi: 10.1016/0092-8674(79)90100-4. [DOI] [PubMed] [Google Scholar]
  42. 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]

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