Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1985 Aug;5(8):2019–2028. doi: 10.1128/mcb.5.8.2019

Regulation of simian virus 40 gene expression in Xenopus laevis oocytes.

T Michaeli, C Prives
PMCID: PMC366920  PMID: 3018545

Abstract

Expression of the simian virus 40 (SV40) early and late regions was examined in Xenopus laevis oocytes microinjected with viral DNA. In contrast to the situation in monkey cells, both late-strand-specific (L-strand) RNA and early-strand-specific (E-strand) RNA could be detected as early as 2 h after injection. At all time points tested thereafter, L-strand RNA was synthesized in excess over E-strand RNA. Significantly greater quantities of L-strand, relative to E-strand, RNA were detected over a 100-fold range of DNA concentrations injected. Analysis of the subcellular distribution of [35S]methionine-labeled viral proteins revealed that while the majority of the VP-1 and all detectable small t antigen were found in the oocyte cytoplasm, most of the large T antigen was located in the oocyte nucleus. The presence of the large T antigen in the nucleus led us to investigate whether this viral product influences the relative synthesis of late or early RNA in the oocyte as it does in infected monkey cells. Microinjection of either mutant C6 SV40 DNA, which encodes a large T antigen unable to bind specifically to viral regulatory sequences, or deleted viral DNA lacking part of the large T antigen coding sequences yielded ratios of L-strand to E-strand RNA that were similar to those observed with wild-type SV40 DNA. Taken together, these observations suggest that the regulation of SV40 RNA synthesis in X. laevis oocytes occurs by a fundamentally different mechanism than that observed in infected monkey cells. This notion was further supported by the observation that the major 5' ends of L-strand RNA synthesized in oocytes were different from those detected in infected cells. Furthermore, only a subset of those L-strand RNAs were polyadenylated.

Full text

PDF
2023

Images in this article

2021Image
on p.2021
2022Image
on p.2022
2023Image
on p.2023
2023Image
on p.2023
2024Image
on p.2024
2025Image
on p.2025
2026Image
on p.2026

Selected References

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

  1. Ben-Ze'ev A., Abulafia R., Aloni Y. SV40 virions and viral RNA metabolism are associated with cellular substructures. EMBO J. 1982;1(10):1225–1231. doi: 10.1002/j.1460-2075.1982.tb00017.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birkenmeier E. H., Chiu N., Radonovich M. F., May E., Salzman N. P. Regulation of simian virus 40 early and late gene transcription without viral DNA replication. J Virol. 1979 Mar;29(3):983–989. doi: 10.1128/jvi.29.3.983-989.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brady J., Bolen J. B., Radonovich M., Salzman N., Khoury G. Stimulation of simian virus 40 late gene expression by simian virus 40 tumor antigen. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2040–2044. doi: 10.1073/pnas.81.7.2040. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dingwall C., Sharnick S. V., Laskey R. A. A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell. 1982 Sep;30(2):449–458. doi: 10.1016/0092-8674(82)90242-2. [DOI] [PubMed] [Google Scholar]
  6. Eppig J. J., Steckman M. L. Comparison of exogenous energy sources for in vitro maintenance of follicle cell-free Xenopus laevis oocytes. In Vitro. 1976 Mar;12(3):173–179. doi: 10.1007/BF02796439. [DOI] [PubMed] [Google Scholar]
  7. Ernoult-Lange M., May E. Evidence of transcription from the late region of the integrated simian virus 40 genome in transformed cells: location of the 5' ends of late transcripts in cells abortively infected and in cells transformed by simian virus 40. J Virol. 1983 Jun;46(3):756–767. doi: 10.1128/jvi.46.3.756-767.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  9. Fradin A., Jove R., Hemenway C., Keiser H. D., Manley J. L., Prives C. Splicing pathways of SV40 mRNAs in X. laevis oocytes differ in their requirements for snRNPs. Cell. 1984 Jul;37(3):927–936. doi: 10.1016/0092-8674(84)90427-6. [DOI] [PubMed] [Google Scholar]
  10. Ghosh P. K., Reddy V. B., Swinscoe J., Lebowitz P., Weissman S. M. Heterogeneity and 5'-terminal structures of the late RNAs of simian virus 40. J Mol Biol. 1978 Dec 25;126(4):813–846. doi: 10.1016/0022-2836(78)90022-0. [DOI] [PubMed] [Google Scholar]
  11. Gluzman Y., Ahrens B. SV40 early mutants that are defective for viral DNA synthesis but competent for transformation of cultured rat and simian cells. Virology. 1982 Nov;123(1):78–92. doi: 10.1016/0042-6822(82)90296-3. [DOI] [PubMed] [Google Scholar]
  12. Graessmann A., Graessmann M., Mueller C. Microinjection of early SV40 DNA fragments and T antigen. Methods Enzymol. 1980;65(1):816–825. doi: 10.1016/s0076-6879(80)65076-9. [DOI] [PubMed] [Google Scholar]
  13. Gurdon J. B., Brown D. D. The transcription of 5 S DNA injected into Xenopus oocytes. Dev Biol. 1978 Dec;67(2):346–356. doi: 10.1016/0012-1606(78)90205-1. [DOI] [PubMed] [Google Scholar]
  14. Gurdon J. B. Changes in somatic cell nuclei inserted into growing and maturing amphibian oocytes. J Embryol Exp Morphol. 1968 Nov;20(3):401–414. [PubMed] [Google Scholar]
  15. Hansen U., Tenen D. G., Livingston D. M., Sharp P. A. T antigen repression of SV40 early transcription from two promoters. Cell. 1981 Dec;27(3 Pt 2):603–613. doi: 10.1016/0092-8674(81)90402-5. [DOI] [PubMed] [Google Scholar]
  16. Harland R. M., Laskey R. A. Regulated replication of DNA microinjected into eggs of Xenopus laevis. Cell. 1980 Oct;21(3):761–771. doi: 10.1016/0092-8674(80)90439-0. [DOI] [PubMed] [Google Scholar]
  17. Hay N., Skolnik-David H., Aloni Y. Attenuation in the control of SV40 gene expression. Cell. 1982 May;29(1):183–193. doi: 10.1016/0092-8674(82)90102-7. [DOI] [PubMed] [Google Scholar]
  18. Hayward G. S. Gel electrophoretic separation of the complementary strands of bacteriophage DNA. Virology. 1972 Jul;49(1):342–344. doi: 10.1016/s0042-6822(72)80042-4. [DOI] [PubMed] [Google Scholar]
  19. Jove R., Sperber D. E., Manley J. L. Transcription of methylated eukaryotic viral genes in a soluble in vitro system. Nucleic Acids Res. 1984 Jun 11;12(11):4715–4730. doi: 10.1093/nar/12.11.4715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. A short amino acid sequence able to specify nuclear location. Cell. 1984 Dec;39(3 Pt 2):499–509. doi: 10.1016/0092-8674(84)90457-4. [DOI] [PubMed] [Google Scholar]
  21. Keller J. M., Alwine J. C. Activation of the SV40 late promoter: direct effects of T antigen in the absence of viral DNA replication. Cell. 1984 Feb;36(2):381–389. doi: 10.1016/0092-8674(84)90231-9. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Lane C. D. The fate of foreign proteins introduced in Xenopus oocytes. Cell. 1981 May;24(2):281–282. doi: 10.1016/0092-8674(81)90315-9. [DOI] [PubMed] [Google Scholar]
  24. Lanford R. E., Butel J. S. Construction and characterization of an SV40 mutant defective in nuclear transport of T antigen. Cell. 1984 Jul;37(3):801–813. doi: 10.1016/0092-8674(84)90415-x. [DOI] [PubMed] [Google Scholar]
  25. Lebowitz P., Weissman S. M. Organization and transcription of the simian virus 40 genome. Curr Top Microbiol Immunol. 1979;87:43–172. doi: 10.1007/978-3-642-67344-3_3. [DOI] [PubMed] [Google Scholar]
  26. Lin W., Hata T., Kasamatsu H. Subcellular distribution of viral structural proteins during simian virus 40 infection. J Virol. 1984 May;50(2):363–371. doi: 10.1128/jvi.50.2.363-371.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Miller T. J., Mertz J. E. Template structural requirements for transcription in vivo by RNA polymerase II. Mol Cell Biol. 1982 Dec;2(12):1595–1607. doi: 10.1128/mcb.2.12.1595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miller T. J., Stephens D. L., Mertz J. E. Kinetics of accumulation and processing of simian virus 40 RNA in Xenopus laevis oocytes injected with simian virus 40 DNA. Mol Cell Biol. 1982 Dec;2(12):1581–1594. doi: 10.1128/mcb.2.12.1581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Piatak M., Subramanian K. N., Roy P., Weissman S. M. Late messenger RNA production by viable simian virus 40 mutants with deletions in the leader region. J Mol Biol. 1981 Dec 15;153(3):589–618. doi: 10.1016/0022-2836(81)90409-5. [DOI] [PubMed] [Google Scholar]
  30. Prives C. L., Shure H. Cell-free translation of simian virus 40 16S and 19S L-strand-specific mRNA classes to simian virus 40 major VP-1 and minor VP-2 and VP-3 capsid proteins. J Virol. 1979 Mar;29(3):1204–1212. doi: 10.1128/jvi.29.3.1204-1212.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Prives C., Barnet B., Scheller A., Khoury G., Jay G. Discrete regions of simian virus 40 large T antigen are required for nonspecific and viral origin-specific DNA binding. J Virol. 1982 Jul;43(1):73–82. doi: 10.1128/jvi.43.1.73-82.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Rio D., Robbins A., Myers R., Tjian R. Regulation of simian virus 40 early transcription in vitro by a purified tumor antigen. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5706–5710. doi: 10.1073/pnas.77.10.5706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rungger D., Türler H. DNAs of simian virus 40 and polyoma direct the synthesis of viral tumor antigens and capsid proteins in Xenopus oocytes. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6073–6077. doi: 10.1073/pnas.75.12.6073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Scheller A., Covey L., Barnet B., Prives C. A small subclass of SV40 T antigen binds to the viral origin of replication. Cell. 1982 Jun;29(2):375–383. doi: 10.1016/0092-8674(82)90154-4. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wickens M. P., Gurdon J. B. Post-transcriptional processing of simian virus 40 late transcripts in injected frog oocytes. J Mol Biol. 1983 Jan 5;163(1):1–26. doi: 10.1016/0022-2836(83)90027-x. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES