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. 1971 Oct;8(4):430–436. doi: 10.1128/jvi.8.4.430-436.1971

Spontaneous Virus Production by Clonal Lines of Simian Virus 40-Transformed Cells and Effects of Superinfection by Deoxyribonucleic Acid from Mutant Simian Virus 40 Strains

D R Dubbs 1, Saul Kit 1
PMCID: PMC376216  PMID: 4331650

Abstract

Small amounts of infectious simian virus 40 (SV40) were recovered from parental cultures of SV40-transformed human embryonic lung (WI38 Va13A) cells, from 12 primary clones, from 17 secondary clones, and from 18 tertiary clones. The cloning experiments demonstrated that the capacity for spontaneous virus production is a hereditary property of WI38 Va13A cells. Infectious virus was not recovered from every clone at every passage. Repeated trials at different passage levels were necessary to detect virus production. Approximately one in 105 to 106 of the cells of the clonal lines initiated plaque formation when plated on the CV-1 line of African green monkey kidney cells. No increase in infectious center formation was observed after the clonal lines were treated with bromodeoxyuridine, iododeoxyuridine, or mitomycin C or after heterokaryon formation of treated cells with CV-1 cells. The clonal lines of WI38 Va13A cells were susceptible to superinfection by SV40 deoxyribonucleic acid (DNA). To determine whether only those cells which spontaneously produced virus supported the replication of superinfecting SV40 DNA, cultures were infected with DNA from a plaque morphology mutant and a temperature-sensitive mutant of SV40. After infection by SV40 DNA, approximately 100 to 4,400 times more transformed cells formed infectious centers than were spontaneously producing virus. To determine whether the resident SV40 genome or the superinfecting SV40 genome was replicating, infectious centers produced by SV40 DNA-infected WI38 Va13A cells on CV-1 monolayers were picked and the progeny virus was analyzed. Only the superinfecting SV40 was recovered from the infectious centers, indicating that in the majority of superinfected cells the resident SV40 was not induced to replicate.

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Selected References

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

  1. Burns W. H., Black P. H. Analysis of simian virus 40-induced transformation of hamster kidney tissue in vitro. V. Variability of virus recovery from cell clones inducible with mitomycin C and cell fusion. J Virol. 1968 Jun;2(6):606–609. doi: 10.1128/jvi.2.6.606-609.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Dubbs D. R., Kit S. Isolation of defective lysogens from Simian virus 40-transformed mouse kidney cultures. J Virol. 1968 Nov;2(11):1272–1282. doi: 10.1128/jvi.2.11.1272-1282.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dubbs D. R., Kit S. Isolation of double lysogens from 3T3 cells transformed by plaque morphology mutants of SV40. Proc Natl Acad Sci U S A. 1970 Mar;65(3):536–543. doi: 10.1073/pnas.65.3.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fogel M., Sachs L. Induction of virus synthesis in polyoma transformed cells by ultraviolet light and mitomycin C. Virology. 1970 Jan;40(1):174–177. doi: 10.1016/0042-6822(70)90391-0. [DOI] [PubMed] [Google Scholar]
  5. Fogel M., Sachs L. The activation of virus synthesis in polyoma-transformed cells. Virology. 1969 Mar;37(3):327–334. doi: 10.1016/0042-6822(69)90216-5. [DOI] [PubMed] [Google Scholar]
  6. GERBER P. VIROGENIC HAMSTER TUMOR CELLS: INDUCTION OF VIRUS SYNTHESIS. Science. 1964 Aug 21;145(3634):833–833. doi: 10.1126/science.145.3634.833. [DOI] [PubMed] [Google Scholar]
  7. GIRARDI A. J., JENSEN F. C., KOPROWSKI H. SV40-INDUCED TRANFORMATION OF HUMAN DIPLOID CELLS: CRISIS AND RECOVERY. J Cell Physiol. 1965 Feb;65:69–83. doi: 10.1002/jcp.1030650110. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Jensen F. C., Koprowski H. Absence of repressor in SV40-transformed cells. Virology. 1969 Apr;37(4):687–690. doi: 10.1016/0042-6822(69)90290-6. [DOI] [PubMed] [Google Scholar]
  11. Kit S., Kurimura T., Brown M., Dubbs D. R. Identification of the simian virus 40 which replicates when simian virus 40-transformed human cells are fused with simian virus 40-transformed mouse cells or superinfected with simian virus 40 deoxyribonucleic acid. J Virol. 1970 Jul;6(1):69–77. doi: 10.1128/jvi.6.1.69-77.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kit S., Kurimura T., Salvi M. L., Dubbs D. R. Activation of infectious SV40 synthesis in transformed cells. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1239–1246. doi: 10.1073/pnas.60.4.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kit S., Tokuno S., Nakajima K., Trkula D., Dubbs D. R. Temperature-sensitive simian virus 40 mutant defective in a late function. J Virol. 1970 Sep;6(3):286–294. doi: 10.1128/jvi.6.3.286-294.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. PUCK T. T., MARCUS P. I., CIECIURA S. J. Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med. 1956 Feb 1;103(2):273–283. doi: 10.1084/jem.103.2.273. [DOI] [PMC free article] [PubMed] [Google Scholar]

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