Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1980 Mar;33(3):1173–1181. doi: 10.1128/jvi.33.3.1173-1181.1980

Isolation and characterization of polyoma uncoating intermediates from the nuclei of infected mouse cells.

V D Winston, J B Bolen, R A Consigli
PMCID: PMC288650  PMID: 6245267

Abstract

A method was developed which enabled the efficient recovery of polyoma uncoating intermediates from the nuclei of infected cells at early times after infection (15 min to 12 h). Cells were infected with radiolabeled virus and lysed with the detergent Nonidet P-40. The nuclei were then collected and sonicated, and the products were analyzed on sucrose gradients. The uncoating intermediate sedimented at 190S and was a viral DNA-protein complex closely associated with a structure of host origin. The host material associated with the 190S uncoating intermediate was determined by polyacrylamide gel electrophoresis and visualized by electron microscopy. The amount of 190S uncoating intermediate found in the nucleus increased with time after infection. The viral DNA was predominantly for I. All of the viral proteins were present in the 190S uncoating intermediate in amounts similar to those found in viral DNA-protein complex cores.

Full text

PDF
1173

Images in this article

1179Image
on p.1179

Selected References

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

  1. Barbanti-Brodano G., Swetly P., Koprowski H. Early events in the infection of permissive cells with simian virus 40: adsorption, penetration, and uncoating. J Virol. 1970 Jul;6(1):78–86. doi: 10.1128/jvi.6.1.78-86.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berezney R., Coffey D. S. The nuclear protein matrix: isolation, structure, and functions. Adv Enzyme Regul. 1976;14:63–100. doi: 10.1016/0065-2571(76)90008-x. [DOI] [PubMed] [Google Scholar]
  3. Brady J. N., Winston V. D., Consigli R. A. Characterization of a DNA-protein complex and capsomere subunits derived from polyoma virus by treatment with ethyleneglycol-bis-N,N'-tetraacetic acid and dithiothreitol. J Virol. 1978 Jul;27(1):193–204. doi: 10.1128/jvi.27.1.193-204.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brady J. N., Winston V. D., Consigli R. A. Dissociation of polyoma virus by the chelation of calcium ions found associated with purified virions. J Virol. 1977 Sep;23(3):717–724. doi: 10.1128/jvi.23.3.717-724.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Comings D. E., Wallack A. S. DNA-binding properties of nuclear matrix proteins. J Cell Sci. 1978 Dec;34:233–246. doi: 10.1242/jcs.34.1.233. [DOI] [PubMed] [Google Scholar]
  6. Consigli R. A., Center M. S. Recent advances in polyoma virus research. CRC Crit Rev Microbiol. 1978;6(3):263–299. doi: 10.3109/10408417809090624. [DOI] [PubMed] [Google Scholar]
  7. Consigli R. A., Zabielski J., Weil R. Plaque assay for polyoma virus on primary mouse kidney cell cultures. Appl Microbiol. 1973 Oct;26(4):627–628. doi: 10.1128/am.26.4.627-628.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dijkwel P. A., Mullenders L. H., Wanka F. Analysis of the attachment of replicating DNA to a nuclear matrix in mammalian interphase nuclei. Nucleic Acids Res. 1979 Jan;6(1):219–230. doi: 10.1093/nar/6.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dingman C. W. Bidirectional chromosome replication: some topological considerations. J Theor Biol. 1974 Jan;43(1):187–195. doi: 10.1016/s0022-5193(74)80052-4. [DOI] [PubMed] [Google Scholar]
  10. Frost E., Bourgaux P. Decapsidation of polyoma virus: identification of subviral species. Virology. 1975 Nov;68(1):245–255. doi: 10.1016/0042-6822(75)90165-8. [DOI] [PubMed] [Google Scholar]
  11. Goldstein D. A., Hall M. R., Meinke W. Properties of nucleoprotein complexes containing replicating polyoma DNA. J Virol. 1973 Oct;12(4):887–900. doi: 10.1128/jvi.12.4.887-900.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Green M. H., Miller H. I., Hendler S. Isolation of a polyoma-nucleoprotein complex from infected mouse-cell cultures. Proc Natl Acad Sci U S A. 1971 May;68(5):1032–1036. doi: 10.1073/pnas.68.5.1032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hobart P., Duncan R., Infante A. A. Association of DNA synthesis with the nuclear membrane in sea urchin embryos. Nature. 1977 Jun 9;267(5611):542–544. doi: 10.1038/267542a0. [DOI] [PubMed] [Google Scholar]
  14. Hummeler K., Tomassini N., Sokol F. Morphological aspects of the uptake of simian virus 40 by permissive cells. J Virol. 1970 Jul;6(1):87–93. doi: 10.1128/jvi.6.1.87-93.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Infante A. A., Firshein W., Hobart P., Murray L. A nuclear membrane-associated DNA complex in cultured mammalian cells capable of synthesizing DNA in vitro. Biochemistry. 1976 Nov 2;15(22):4810–4817. doi: 10.1021/bi00667a010. [DOI] [PubMed] [Google Scholar]
  16. LeBlanc D. J., Singer M. F. Localization of replicating DNA of simian virus 40 in monkey kidney cells. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2236–2240. doi: 10.1073/pnas.71.6.2236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. MacGregor J. P., Chen Y. H., Goldstein D. A., Hall M. R. Properties of the histones associated with simian virus 40 replicative form nucleoprotein complex. Biochem Biophys Res Commun. 1978 Aug 14;83(3):814–821. doi: 10.1016/0006-291x(78)91467-5. [DOI] [PubMed] [Google Scholar]
  18. Mackay R. L., Consigli R. A. Early events in polyoma virus infection: attachment, penetration, and nuclear entry. J Virol. 1976 Aug;19(2):620–636. doi: 10.1128/jvi.19.2.620-636.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. McMillen J., Center M. S., Consigli R. A. Origin of the polyoma virus-associated endonuclease. J Virol. 1975 Jan;17(1):127–131. doi: 10.1128/jvi.17.1.127-131.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. McMillen J., Consigli R. A. Immunological reactivity of antisera to sodium dodecyl sulfate-derived polypeptides of polyoma virions. J Virol. 1977 Mar;21(3):1113–1120. doi: 10.1128/jvi.21.3.1113-1120.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Riley D. E., Keller J. M. Cell cycle-dependent changes in non-membranous nuclear ghosts from HeLa cells. J Cell Sci. 1978 Feb;29:129–146. doi: 10.1242/jcs.29.1.129. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Sinha A. A., Mizuno N. S. DNA synthesis associated with a DNA-nuclear-membrane complex from rat liver. Cell Tissue Res. 1977 Sep 26;183(2):191–201. doi: 10.1007/BF00226619. [DOI] [PubMed] [Google Scholar]
  24. Smith G. L., Consigli R. A. Transient inhibition of polyoma virus synthesis by Sendai virus (parainfluenza I). I. Demonstration and nature of the inhibition by inactivated virus. J Virol. 1972 Dec;10(6):1091–1097. doi: 10.1128/jvi.10.6.1091-1097.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tremblay G. Y., Daniels M. J., Schaechter M. Isolation of a cell membrane-DNA-nascent RNA complex from bacteria. J Mol Biol. 1969 Feb 28;40(1):65–76. doi: 10.1016/0022-2836(69)90296-4. [DOI] [PubMed] [Google Scholar]
  26. Yang Y. C., Hearing P., Rundell K. Cellular proteins associated with simian virus 40 early gene products in newly infected cells. J Virol. 1979 Oct;32(1):147–154. doi: 10.1128/jvi.32.1.147-154.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES