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. 1980 Jul;35(1):141–149. doi: 10.1128/jvi.35.1.141-149.1980

Preferential replication of a class of host-substituted defective simian virus 40 variants at low temperature.

M F Singer, R E Thayer
PMCID: PMC288789  PMID: 6251262

Abstract

The host-substituted variant termed CVP8/1/P2 (EcoRI res) was first isolated several years ago after serial passage of simian virus 40 strain 777 on BSC-1 cells at 37 degrees C. When BSC-1 are coinfected with wild-type simian virus 40 strain 777 and variant CVP8/1/P2 (EcoRI res), the variant rapidly becomes the dominant species produced, often representing as much as 80% of the total DNA I synthesized after infection. We present evidence that the replicative advantage of the variant was increased when the infection was carried out at 33 rather than 37 degrees C. Also described are nine new and independent serial passage experiments carried out at 33 degrees C with several purified wild-type virus stocks, including strain 776, and both BSC-1 and primary African green monkey kidney cells. In each series variants related to CVPs/1/P2 (EcoRI res) were detected in the progeny viral genomes after four serial passages. Hybridization data suggest that at least some of these variant DNA I molecules contain simian virus 40 DNA sequences, monkey alpha-component DNA sequences (highly repetitive), and the infrequently reiterated monkey DNA sequences found in CVP8/1/P2 (EcoRI res), all covalently linked as in CPV8/1/P2 (EcoRI res). It appears that this type of variant emerges with some frequency during infection and is then preferentially replicated at 33 degrees C, thereby becoming readily detectable in passaged stocks. A variety of control experiments indicated that the repeated emergence of similar, if not identical, variants is unlikely to be the result of inadvertent cross-contamination or the presence of detectable amounts of the variant in the plaque-purified viral stocks.

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Selected References

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

  1. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  2. Britten R. J., Graham D. E., Neufeld B. R. Analysis of repeating DNA sequences by reassociation. Methods Enzymol. 1974;29:363–418. doi: 10.1016/0076-6879(74)29033-5. [DOI] [PubMed] [Google Scholar]
  3. Fareed G. C., Davoli D. Molecular biology of papovaviruses. Annu Rev Biochem. 1977;46:471–522. doi: 10.1146/annurev.bi.46.070177.002351. [DOI] [PubMed] [Google Scholar]
  4. Fiers W., Contreras R., Haegemann G., Rogiers R., Van de Voorde A., Van Heuverswyn H., Van Herreweghe J., Volckaert G., Ysebaert M. Complete nucleotide sequence of SV40 DNA. Nature. 1978 May 11;273(5658):113–120. doi: 10.1038/273113a0. [DOI] [PubMed] [Google Scholar]
  5. Frenkel N., Lavi S., Winocour E. The host DNA sequences in different populations of serially passaged SV40. Virology. 1974 Jul;60(1):9–20. doi: 10.1016/0042-6822(74)90360-2. [DOI] [PubMed] [Google Scholar]
  6. Gutai M. W., Nathans D. Evolutionary variants of simian virus 40: Cellular DNA sequences and sequences at recombinant joints of substituted variants. J Mol Biol. 1978 Dec 5;126(2):275–288. doi: 10.1016/0022-2836(78)90363-7. [DOI] [PubMed] [Google Scholar]
  7. Gutai M. W., Nathans D. Evolutionary variants of simian virus 40: Nucleotide sequence of a conserved SV40 DNA segment containing the origin of viral DNA replication as an inverted repetition. J Mol Biol. 1978 Dec 5;126(2):259–274. doi: 10.1016/0022-2836(78)90362-5. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Kay A. C., Rao G. R., Singer M. F. Comparison of two viable variants of simian virus 40. J Virol. 1978 Jan;25(1):339–348. doi: 10.1128/jvi.25.1.339-348.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kelly T. J., Jr, Nathans D. The genome of simian virus 40. Adv Virus Res. 1977;21:85–173. doi: 10.1016/s0065-3527(08)60762-9. [DOI] [PubMed] [Google Scholar]
  12. Kuff E. L., Ferdinand F. J., Khoury G. Transcription of host-substituted simian virus 40 DNA in whole cells and extracts. J Virol. 1978 Jan;25(1):28–36. doi: 10.1128/jvi.25.1.28-36.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lavi S., Rozenblatt S., Singer M. F., Winocour E. Acquisition of sequences homologous to host DNA by closed circular simian virus 40 DNA. II. Further studies on the serial passage of virus clones. J Virol. 1973 Sep;12(3):492–500. doi: 10.1128/jvi.12.3.492-500.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lavi S., Winocour E. Acquisition of sequences homologous to host deoxyribonucleic acid by closed circular simian virus 40 deoxyribonucleic acid. J Virol. 1972 Feb;9(2):309–316. doi: 10.1128/jvi.9.2.309-316.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lee T. N., Brockman W. W., Nathans D. Evolutionary variants of simian virus 40: cloned substituted variants containing multiple initiation sites for DNA replication. Virology. 1975 Jul;66(1):53–69. doi: 10.1016/0042-6822(75)90178-6. [DOI] [PubMed] [Google Scholar]
  16. Lee T. N., Nathans D. Evolutionary variants of simian virus 40: replication and encapsidation of variant DNA. Virology. 1979 Jan 30;92(2):291–298. doi: 10.1016/0042-6822(79)90134-x. [DOI] [PubMed] [Google Scholar]
  17. Maio J. J. DNA strand reassociation and polyribonucleotide binding in the African green monkey, Cercopithecus aethiops. J Mol Biol. 1971 Mar 28;56(3):579–595. doi: 10.1016/0022-2836(71)90403-7. [DOI] [PubMed] [Google Scholar]
  18. Martin M. A., Gelb L. D., Fareed G. C., Milstien J. B. Reassortment of simian virus 40 DNA during serial undiluted passage. J Virol. 1973 Oct;12(4):748–757. doi: 10.1128/jvi.12.4.748-757.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McCutchan T., Singer M., Rosenberg M. Structure of simian virus 40 recombinants that contain both host and viral DNA sequences. II. The structure of variant 1103 and its comparison to variant CVPS/1P2 (EcoRI res). J Biol Chem. 1979 May 10;254(9):3592–3597. [PubMed] [Google Scholar]
  20. Oren M., Kuff E. L., Winocour E. The presence of common host sequences in different populations of substituted SV40 DNA. Virology. 1976 Sep;73(2):419–430. doi: 10.1016/0042-6822(76)90403-7. [DOI] [PubMed] [Google Scholar]
  21. Oren M., Lavi S., Winocour E. The structure of a cloned substituted SV40 genome. Virology. 1978 Apr;85(2):404–421. doi: 10.1016/0042-6822(78)90448-8. [DOI] [PubMed] [Google Scholar]
  22. Rao G. R., Singer M. F. Studies on a defective variant of simian virus 40 that is substituted with DNA sequences derived from monkey DNA. I. Origin, properties, and purification. J Biol Chem. 1977 Jul 25;252(14):5115–5123. [PubMed] [Google Scholar]
  23. Rao G. R., Singer M. F. Studies on a defective variant of simian virus 40 that is substituted with DNA sequences derived from monkey. II. Structure of DNA. J Biol Chem. 1977 Jul 25;252(14):5124–5134. [PubMed] [Google Scholar]
  24. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Rosenberg H., Singer M., Rosenberg M. Highly reiterated sequences of SIMIANSIMIANSIMIANSIMIANSIMIAN. Science. 1978 Apr 28;200(4340):394–402. doi: 10.1126/science.205944. [DOI] [PubMed] [Google Scholar]
  27. Rosenberg M., Segal S., Kuff E. L., Singer M. F. The nucleotide sequence of repetitive monkey DNA found in defective simian virus 40. Cell. 1977 Aug;11(4):845–857. doi: 10.1016/0092-8674(77)90296-3. [DOI] [PubMed] [Google Scholar]
  28. Rozenblatt S., Lavi S., Singer M. F., Winocour E. Acquisition of sequences homologous to host DNA by closed circular simian virus 40 DNA. 3. Host sequences. J Virol. 1973 Sep;12(3):501–510. doi: 10.1128/jvi.12.3.501-510.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Segal S., Garner M., Singer M. F., Rosenberg M. In situ hybridization of repetitive monkey genome sequences isolated from defective simian virus 40 DNA. Cell. 1976 Oct;9(2):247–257. doi: 10.1016/0092-8674(76)90116-1. [DOI] [PubMed] [Google Scholar]
  30. Singer D. S. Arrangement of a highly repeated DNA sequence in the genome and chromatin of the African green monkey. J Biol Chem. 1979 Jun 25;254(12):5506–5514. [PubMed] [Google Scholar]
  31. Villarreal L. P., Berg P. Hybridization in situ of SV40 plaques: detection of recombinant SV40 virus carrying specific sequences of nonviral DNA. Science. 1977 Apr 8;196(4286):183–185. doi: 10.1126/science.191907. [DOI] [PubMed] [Google Scholar]
  32. Villarreal L. P., White R. T., Berg P. Mutational alterations within the simian virus 40 leader segment generate altered 16S and 19S mRNA's. J Virol. 1979 Jan;29(1):209–219. doi: 10.1128/jvi.29.1.209-219.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wakamiya T., McCutchan T., Rosenberg M., Singer M. Structure of simian virus 40 recombinants that contain both host and viral DNA sequences. I. The structure of variant CVPS/1/P2 (EcoRI res). J Biol Chem. 1979 May 10;254(9):3584–3591. [PubMed] [Google Scholar]
  34. Zasloff M., Ginder G. D., Felsenfeld G. A new method for the purification and identification of covalently closed circular DNA molcules. Nucleic Acids Res. 1978 Apr;5(4):1139–1152. doi: 10.1093/nar/5.4.1139. [DOI] [PMC free article] [PubMed] [Google Scholar]

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