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. 1978 Mar;25(3):719–729. doi: 10.1128/jvi.25.3.719-729.1978

Polyoma virus-specific RNA synthesis in an inducible line of polyoma virus-transformed rat cells.

H Manor, R Kamen
PMCID: PMC525965  PMID: 205669

Abstract

Viral RNA present in the inducible LPT clone 1A of polyoma virus-transformed rat cells was characterized before and after mitomycin C induction by hybridization with 32P-labeled separated E and L strands of polyoma viral DNA restriction endonuclease fragments. In clone 1A cells maintained under normal growth conditions, the cytoplasm contained a transcript of the E-strand DNA from the "early" region similar to that previously identified in lytically infected cells, as well as minor quantities of RNA complementary to less than one-half of the L- and the E-strand DNA from the "late" region. Nuclei of normally growing cells contained the same species found in the cytoplasm, as well as an additional abundant RNA complementary to one-half of the L-strand DNA of the late region. No significant changes occurred in the cytoplasmic viral RNA after mitomycin C treatment before the onset of viral DNA replication, but the concentration of the nuclear L-strand DNA transcript diminished. After the onset of viral DNA replication after mitomycin C treatment, transcripts of virtually the entire L-strand DNA were found in the nuclei, and a 10-fold increase was observed in the abundance of RNA transcribed from the E strand of the early region. In the cytoplasm, the abundance of the early RNA increased about 25-fold and late RNA complementary to the L-strand DNA of the late region was found in a similar quantity. The synthesis of both the early and the late RNA species was inhibited if viral DNA replication was blocked with 5-fluorodeoxyuridine. We conclude that the induction of viral DNA replication in LPT cells is not determined at the level of mRNA synthesis.

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Selected References

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

  1. Bacheler L. T. Virus-secific transcription in 3T3 cells transformed by the ts-a mutant of polyoma virus. J Virol. 1977 Apr;22(1):54–64. doi: 10.1128/jvi.22.1.54-64.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Crawford L. V., Robbins A. K. The cleavage of polyoma virus DNA by restriction enzymes KpnI and PstI. J Gen Virol. 1976 Jun;31(3):315–321. doi: 10.1099/0022-1317-31-3-315. [DOI] [PubMed] [Google Scholar]
  3. DULBECCO R., VOGT M. Plaque formation and isolation of pure lines with poliomyelitis viruses. J Exp Med. 1954 Feb;99(2):167–182. doi: 10.1084/jem.99.2.167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fogel M. Induction of virus synthesis in polyoma-transformed cells by DNA antimetabolites and by irradiation after pretreatment with 5-bromodeoxyuridine. Virology. 1972 Jul;49(1):12–22. doi: 10.1016/s0042-6822(72)80003-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Griffin B. E., Fried M. Amplification of a specific region of the polyoma virus genome. Nature. 1975 Jul 17;256(5514):175–179. doi: 10.1038/256175a0. [DOI] [PubMed] [Google Scholar]
  8. Griffin B. E., Fried M., Cowie A. Polyoma DNA: a physical map. Proc Natl Acad Sci U S A. 1974 May;71(5):2077–2081. doi: 10.1073/pnas.71.5.2077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hudson J., Goldstein D., Weil R. A study on the transcription of the polyoma viral genome. Proc Natl Acad Sci U S A. 1970 Jan;65(1):226–233. doi: 10.1073/pnas.65.1.226. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kamen R., Lindstrom D. M., Shure H., Old R. W. Virus-specific RNA in cells productively infected or transformed by polyoma virus. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):187–198. doi: 10.1101/sqb.1974.039.01.025. [DOI] [PubMed] [Google Scholar]
  11. Kamen R., Sedat J., Ziff E. Orientation of the complementary strands of polyoma virus DNA with respect to the DNA physical map. J Virol. 1975 Jan;17(1):212–218. doi: 10.1128/jvi.17.1.212-218.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kamen R., Shure H. Topography of polyoma virus messenger RNA molecules. Cell. 1976 Mar;7(3):361–371. doi: 10.1016/0092-8674(76)90165-3. [DOI] [PubMed] [Google Scholar]
  13. Kelly R. B., Cozzarelli N. R., Deutscher M. P., Lehman I. R., Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. XXXII. Replication of duplex deoxyribonucleic acid by polymerase at a single strand break. J Biol Chem. 1970 Jan 10;245(1):39–45. [PubMed] [Google Scholar]
  14. Lindberg U., Darnell J. E. SV40-specific RNA in the nucleus and polyribosomes of transformed cells. Proc Natl Acad Sci U S A. 1970 Apr;65(4):1089–1096. doi: 10.1073/pnas.65.4.1089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Manor H., Fogel M., Sachs L. Integration of viral into chromosomal deoxyribonucleic acid in an inducible line of polyoma-transformed cells. Virology. 1973 May;53(1):174–185. doi: 10.1016/0042-6822(73)90476-5. [DOI] [PubMed] [Google Scholar]
  16. Manor H., Neer A. Effects of cycloheximide on virus RNA replication in an inducible line of polyoma-transformed rat cells. Cell. 1975 Jul;5(3):311–318. doi: 10.1016/0092-8674(75)90106-3. [DOI] [PubMed] [Google Scholar]
  17. Neer A., Baran N., Manor H. In situ hybridization analysis of polyoma DNA replication in an inducible line of polyoma-transformed cells. Cell. 1977 May;11(1):65–71. doi: 10.1016/0092-8674(77)90317-8. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Prasad I., Zouzias D., Basilico C. State of the viral DNA in rat cells transformed by polyoma virus. I. Virus rescue and the presence of nonintergrated viral DNA molecules. J Virol. 1976 May;18(2):436–444. doi: 10.1128/jvi.18.2.436-444.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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