Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1978 Jan;25(1):28–36. doi: 10.1128/jvi.25.1.28-36.1978

Transcription of host-substituted simian virus 40 DNA in whole cells and extracts.

E L Kuff, F J Ferdinand, G Khoury
PMCID: PMC353897  PMID: 202741

Abstract

Viral transcriptional complexes were extracted from the nuclei of monkey kidney cells infected with wild-type simian virus 40 (SV40) or a variant strain containing a high proportion of host-substituted DNA molecules. The RNAs synthesized by these complexes in an in vitro system were analyzed for their content of SV40 and host sequences by a technique of sequential hybridization to plaque-purified and substituted viral DNAs. The relative labeling of the two types of sequences was commensurate with their proportion in the viral DNA (about 20% host). The substituted virus contains both reiterated and unique types of cellular sequences, and both kinds appeared to be transcribed. Transcripts of the substituted sequences formed a much smaller proportion of the virus related RNA recovered from intact infected cells, suggesting that host sequence transcripts are synthesized but rapidly degraded in the whole cell. The alternative, that transcription of these sequences is artificially enhanced in the in vitro system, cannot be rigorously excluded. We compared the self-annealing of viral RNAs from nuclear extracts of cells infected with wild-type and substituted viruses; transcripts labeled both in vivo and in vitro showed a two- to threefold-higher level of self-annealing in the case of the variant than in the case of wild type SV40.

Full text

PDF
28

Selected References

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

  1. Brockman W. W., Lee T. N., Nathans D. The evolution of new species of viral DNA during serial passage of simian virus 40 at high multiplicity. Virology. 1973 Aug;54(2):384–397. doi: 10.1016/0042-6822(73)90151-7. [DOI] [PubMed] [Google Scholar]
  2. Davoli D., Ganem D., Nussbaum A. L., Fareed G., Howley P. M., Khoury G., Martin M. A. Genome Structures of reiteration mutants of simian virus 40. Virology. 1977 Mar;77(1):110–124. doi: 10.1016/0042-6822(77)90411-1. [DOI] [PubMed] [Google Scholar]
  3. Ferdinand F. J., Brown M., Khoury G. Synthesis and characterization of late lytic simian virus 40 RNA from transcriptional complexes. Virology. 1977 May 1;78(1):150–161. doi: 10.1016/0042-6822(77)90087-3. [DOI] [PubMed] [Google Scholar]
  4. Folk W. R., Fishel B. R. Tandem repetition of the origin of DNA replication in defective polyoma virus DNA's. Virology. 1975 Apr;64(2):447–453. doi: 10.1016/0042-6822(75)90122-1. [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. Fried M., Griffin B. E., Lund E., Robberson D. L. Polyoma virus--a study of wild-type, mutant and defective DNAs. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):45–52. doi: 10.1101/sqb.1974.039.01.009. [DOI] [PubMed] [Google Scholar]
  7. GERBER P. An infectious deoxyribonucleic acid derived from vacuolating virus (SV40). Virology. 1962 Jan;16:96–97. doi: 10.1016/0042-6822(62)90209-x. [DOI] [PubMed] [Google Scholar]
  8. Ganem D., Nussbaum A. L., Davoli D., Fareed G. C. Propagation of a segment of bacteriophage lamda-DNA in monkey cells after covalent linkage to a defective simian virus 40 genome. Cell. 1976 Mar;7(3):349–359. doi: 10.1016/0092-8674(76)90164-1. [DOI] [PubMed] [Google Scholar]
  9. Gariglio P., Mousset S. Isolation and partial characterization of a nuclear RNA polymerase - SV40 DNA complex. FEBS Lett. 1975 Aug 1;56(1):149–155. doi: 10.1016/0014-5793(75)80130-x. [DOI] [PubMed] [Google Scholar]
  10. Gillespie D., Spiegelman S. A quantitative assay for DNA-RNA hybrids with DNA immobilized on a membrane. J Mol Biol. 1965 Jul;12(3):829–842. doi: 10.1016/s0022-2836(65)80331-x. [DOI] [PubMed] [Google Scholar]
  11. Goff S. P., Berg P. Construction of hybrid viruses containing SV40 and lambda phage DNA segments and their propagation in cultured monkey cells. Cell. 1976 Dec;9(4 Pt 2):695–705. doi: 10.1016/0092-8674(76)90133-1. [DOI] [PubMed] [Google Scholar]
  12. Green M. H., Brooks T. L. Isolation of two forms of SV40 nucleoprotein containing RNA polymerase from infected monkey cells. Virology. 1976 Jul 1;72(1):110–120. doi: 10.1016/0042-6822(76)90316-0. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hamer D. H., Davoli D., Thomas C. A., Jr, Fareed G. C. Simian virus 40 carrying an Escherichia coli suppressor gene. J Mol Biol. 1977 May 15;112(2):155–182. doi: 10.1016/s0022-2836(77)80137-x. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Khoury G., Fareed G. C., Berry K., Martin M. A., Lee T. N., Nathans D. Characterization of a rearrangement in viral DNA: mapping of the circular simian virus 40-like DNA containing a triplication of a specific one-third of the viral genome. J Mol Biol. 1974 Aug 5;87(2):289–301. doi: 10.1016/0022-2836(74)90150-8. [DOI] [PubMed] [Google Scholar]
  17. Khoury G., May E. Regulation of early and late simian virus 40 transcription: overproduction of early viral RNA in the absence of a functional T-antigen. J Virol. 1977 Jul;23(1):167–176. doi: 10.1128/jvi.23.1.167-176.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Laub O., Aloni Y. Transcription of simian virus 40. V. Regulattion of simian virus 40 gene expression. J Virol. 1975 Nov;16(5):1171–1183. doi: 10.1128/jvi.16.5.1171-1183.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. 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]
  22. 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]
  23. Nussbaum A. L., Davoli D., Ganem D., Fareed G. C. Construction and propagation of a defective simian virus 40 genome bearing an operator from bacteriophage lambda. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1068–1072. doi: 10.1073/pnas.73.4.1068. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Penman S. RNA metabolism in the HeLa cell nucleus. J Mol Biol. 1966 May;17(1):117–130. doi: 10.1016/s0022-2836(66)80098-0. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Reed S. I., Stark G. R., Alwine J. C. Autoregulation of simian virus 40 gene A by T antigen. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3083–3087. doi: 10.1073/pnas.73.9.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Risser R., Mulder C. Relative locations of rearrangements in the DNA of defective simian virus 40 (SV40). Virology. 1974 Apr;58(2):424–438. doi: 10.1016/0042-6822(74)90077-4. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Shmookler R. J., Buss J., Green M. H. Properties of the polyoma virus transcription complex obtained from mouse nuclei. Virology. 1974 Jan;57(1):122–127. doi: 10.1016/0042-6822(74)90113-5. [DOI] [PubMed] [Google Scholar]
  31. Weinberg R. A., Warnaar S. O., Winocour E. Isolation and characterization of simian virus 40 ribonucleic acid. J Virol. 1972 Aug;10(2):193–201. doi: 10.1128/jvi.10.2.193-201.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Winocour E., Frenkel N., Lavi S., Osenholts M., Rozenblatt S. Host substitution in SV40 and polyoma DNA. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):101–108. doi: 10.1101/sqb.1974.039.01.015. [DOI] [PubMed] [Google Scholar]

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

RESOURCES