Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1983 Aug;47(2):344–353. doi: 10.1128/jvi.47.2.344-353.1983

The adenovirus type 2-simian virus 40 hybrid virus Ad2+ND4 requires deletion variants to grow in monkey cells.

A M Lewis Jr, H Westphal
PMCID: PMC255266  PMID: 6312070

Abstract

The Ad2+ND4 virus is an adenovirus type 2 (Ad2)-simian virus 40 (SV40) recombination. The Ad2 genome of this recombinant has a rearrangement within early region 3; Ad2 DNA sequences between map positions 81.3 and 85.5 have been deleted, and the SV40 DNA sequences between map positions 0.11 and 0.626 have been inserted into the deletion in an 81.3-0.626 orientation. Nonhybrid Ad2 is defective in monkey cells; however, the Ad2+ND4 virus can replicate in monkey cells due to the expression of the SV40-enhancing function encoded by the DNA insert. Stocks of the Ad2+ND4 hybrid were produced in primary monkey cells by using the progeny of a three-step plaque purification procedure and were considered to be homogeneous populations of Ad2+ND4 virions because they induced plaques in primary monkey cells by first-order kinetics. By studying the kinetics of plaque induction in continuous lines (BSC-1 and CV-1) of monkey cells, we have found that stocks (prepared with virions before and after plaque purification) of Ad2+ND4 are actually heterogeneous populations of Ad2+ND4 virions and Ad2+ND4 deletion variants that lack SV40 and frequently Ad2 DNA sequences at the left Ad2-SV40 junction. Due to the defectiveness of the Ad2+ND4 virus, the production of progeny in BSC-1 and CV-1 cells requires complementation between the Ad2+ND4 genome and the genome of an Ad2+ND4 deletion variant. Since the deletion variants that have been obtained from Ad2+ND4 stocks do not express the SV40-enhancing function in that they cannot produce progeny in monkey cells, we conclude that they are providing an Ad2 component that is essential for the production of Ad2+ND4 progeny. These data imply that the Ad2+ND4 virus is incapable of replicating in singly infected primary monkey cells without generating deletion variants that are missing various amounts of DNA around the left Ad2-SV40 junction in the hybrid genome. As the deletion variants that arise from the Ad2+ND4 virus are created by nonhomologous DNA recombination, the generation of deletion variants in monkey cells infected with Ad2+ND4 may be a useful model for studying this process.

Full text

PDF
344

Images in this article

348Image
on p.348
349Image
on p.349
351Image
on p.351

Selected References

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

  1. Boeyé A., Melnick J. L., Rapp F. SV40-adenovirus "hybrids": presence of two genotypes and the requirement of their complementation for viral replication. Virology. 1966 Jan;28(1):56–70. doi: 10.1016/0042-6822(66)90306-0. [DOI] [PubMed] [Google Scholar]
  2. Fey G., Lewis J. B., Grodzicker T., Bothwell A. Characterization of a fused protein specified by the adenovirus type 2-simian virus 40 hybrid Ad2+ND1 dp2. J Virol. 1979 Apr;30(1):201–217. doi: 10.1128/jvi.30.1.201-217.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Giard D. J., Aaronson S. A., Todaro G. J., Arnstein P., Kersey J. H., Dosik H., Parks W. P. In vitro cultivation of human tumors: establishment of cell lines derived from a series of solid tumors. J Natl Cancer Inst. 1973 Nov;51(5):1417–1423. doi: 10.1093/jnci/51.5.1417. [DOI] [PubMed] [Google Scholar]
  4. Gluzman Y., Van Doren K. Palindromic adenovirus type 5-simian virus 40 hybrid. J Virol. 1983 Jan;45(1):91–103. doi: 10.1128/jvi.45.1.91-103.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grodzicker T., Lewis J. B., Anderson C. W. Conditional lethal mutants of adenovirus type 2-simian virus 40 hybrids. II. Ad2+ND1 host-range mutants that synthesize fragments of the Ad2+ND1 30K protein. J Virol. 1976 Aug;19(2):559–571. doi: 10.1128/jvi.19.2.559-571.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HOPPS H. E., BERNHEIM B. C., NISALAK A., TJIO J. H., SMADEL J. E. BIOLOGIC CHARACTERISTICS OF A CONTINUOUS KIDNEY CELL LINE DERIVED FROM THE AFRICAN GREEN MONKEY. J Immunol. 1963 Sep;91:416–424. [PubMed] [Google Scholar]
  7. JENSEN F. C., GIRARDI A. J., GILDEN R. V., KOPROWSKI H. INFECTION OF HUMAN AND SIMIAN TISSUE CULTURES WITH ROUS SARCOMA VIRUS. Proc Natl Acad Sci U S A. 1964 Jul;52:53–59. doi: 10.1073/pnas.52.1.53. [DOI] [PMC free article] [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., Alwine J., Goldman N., Gruss P., Jay G. New chimeric splice junction in adenovirus type 2-simian virus 40 hybrid viral mRNA. J Virol. 1980 Oct;36(1):143–151. doi: 10.1128/jvi.36.1.143-151.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lebowitz P., Kelly T. J., Jr, Nathans D., Lee T. N., Lewis A. M., Jr A colinear map relating the simian virus 40 (SV40) DNA segments of six adenovirus-SV40 hybrids to the DNA fragments produced by restriction endonuclease cleavage of SV40 DNA. Proc Natl Acad Sci U S A. 1974 Feb;71(2):441–445. doi: 10.1073/pnas.71.2.441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Levine A. S., Levin M. J., Oxman M. N., Lewis A. M., Jr Studies of nondefective adenovirus 2-simian virus 40 hybrid viruses. VII. Characterization of the simian virus 40 RNA species induced by five nondefective hybrid viruses. J Virol. 1973 May;11(5):672–681. doi: 10.1128/jvi.11.5.672-681.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Lewis A. M., Jr, Rowe W. P. Isolation of two plaque variants from the adenovirus type 2-simian virus 40 hybrid population which differ in their efficiency in yielding simian virus 40. J Virol. 1970 Apr;5(4):413–420. doi: 10.1128/jvi.5.4.413-420.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Ling L. E., Manos M. M., Gluzman Y. Sequence of the junction in adenovirus 2-SV40 hybrids: examples of illegitimate recombination. Nucleic Acids Res. 1982 Dec 20;10(24):8099–8112. doi: 10.1093/nar/10.24.8099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. 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]
  18. Rowe W. P., Bau S. G. Studies of adenovirus SV40 hybrid viruses. II. Defectiveness of the hybrid particles. J Exp Med. 1965 Nov 1;122(5):955–966. doi: 10.1084/jem.122.5.955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Westphal H. Genomic arrangement of an adenovirus-simian virus 40 hybrid virus, Ad2+ND4del. J Virol. 1981 Nov;40(2):526–532. doi: 10.1128/jvi.40.2.526-532.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. Zain B. S., Roberts R. J. Characterization and sequence analysis of a recombination site in the hybrid virus Ad2+ND. J Mol Biol. 1978 Mar 25;120(1):13–31. doi: 10.1016/0022-2836(78)90293-0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES