Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1981 Oct;78(10):6081–6085. doi: 10.1073/pnas.78.10.6081

Use of whole-cell fixation to visualize replicating and maturing simian virus 40: identification of new viral gene product.

V Jackson, R Chalkley
PMCID: PMC348981  PMID: 6273846

Abstract

Formaldehyde fixation of simian virus 40 (SV40)-infected CV-1 cells at appropriate times after infection permits us to isolate crosslinked complexes of SV40 minichromosomes during the time of DNA replication and during packaging with viral proteins. Such crosslinked complexes can be separated on the basis of density on CsCl/guanidine . HCl density gradients. During the course of these studies we observed the presence of a low molecular weight protein in a region of the gradient much enriched with viral nucleoproteins. This protein is present only in infected cells and has a molecular weight and amino acid composition consistent with it being the product of the so-called SV40 agnogene.

Full text

PDF
6081

Images in this article

6082Image
on p.6082
6083Image
on p.6083
6084Image
on p.6084
6084Image
on p.6084

Selected References

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

  1. Bloemendal H. The vertebrate eye lens. Science. 1977 Jul 8;197(4299):127–138. doi: 10.1126/science.877544. [DOI] [PubMed] [Google Scholar]
  2. Coca-Prados M., Hsu M. T. Intracellular forms of simian virus 40 nucleoprotein complexes. II. Biochemical and electron microscopic analysis of simian virus 40 virion assembly. J Virol. 1979 Jul;31(1):199–208. doi: 10.1128/jvi.31.1.199-208.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Fanning E., Baumgartner I. Role of fast-sedimenting SV40 nucleoprotein complexes in virus assembly. Virology. 1980 Apr 15;102(1):1–12. doi: 10.1016/0042-6822(80)90064-1. [DOI] [PubMed] [Google Scholar]
  4. Fareed G. C., Davoli D. Molecular biology of papovaviruses. Annu Rev Biochem. 1977;46:471–522. doi: 10.1146/annurev.bi.46.070177.002351. [DOI] [PubMed] [Google Scholar]
  5. Garber E. A., Seidman M. M., Levine A. J. Intracellular SV40 nucleoprotein complexes: synthesis to encapsidation. Virology. 1980 Dec;107(2):389–401. doi: 10.1016/0042-6822(80)90306-2. [DOI] [PubMed] [Google Scholar]
  6. Ghosh P. K., Reddy V. B., Swinscoe J., Choudary P. V., Lebowitz P., Weissman S. M. The 5'-terminal leader sequence of late 16 S mRNA from cells infected with simian virus 40. J Biol Chem. 1978 May 25;253(10):3643–3647. [PubMed] [Google Scholar]
  7. Hunter T. Functional characterization of the early and late mRNAs of simian virus 40. Virology. 1979 Jun;95(2):511–522. doi: 10.1016/0042-6822(79)90505-1. [DOI] [PubMed] [Google Scholar]
  8. Jackson V., Chalkley R. A new method for the isolation of replicative chromatin: selective deposition of histone on both new and old DNA. Cell. 1981 Jan;23(1):121–134. doi: 10.1016/0092-8674(81)90277-4. [DOI] [PubMed] [Google Scholar]
  9. Jackson V., Chalkley R. A reevaluation of new histone deposition on replicating chromatin. J Biol Chem. 1981 May 25;256(10):5095–5103. [PubMed] [Google Scholar]
  10. Jakobovits E. B., Aloni Y. Isolation and characterization of various forms of simian virus 40 DNA-protein complexes. Virology. 1980 Apr 15;102(1):107–118. doi: 10.1016/0042-6822(80)90074-4. [DOI] [PubMed] [Google Scholar]
  11. Jay G., Nomura S., Anderson C. W., Khoury G. Identification of the SV40 agnogene product: a DNA binding protein. Nature. 1981 May 28;291(5813):346–349. doi: 10.1038/291346a0. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  14. Lazarides E., Files J. G., Weber K. Simian virus 40 structural proteins: amino-terminal sequence of the major capsid protein. Virology. 1974 Aug;60(2):584–587. doi: 10.1016/0042-6822(74)90353-5. [DOI] [PubMed] [Google Scholar]
  15. Mertz J. E., Berg P. Viable deletion mutants of simian virus 40: selective isolation by means of a restriction endonuclease from Hemophilus parainfluenzae. Proc Natl Acad Sci U S A. 1974 Dec;71(12):4879–4883. doi: 10.1073/pnas.71.12.4879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Reddy V. B., Thimmappaya B., Dhar R., Subramanian K. N., Zain B. S., Pan J., Ghosh P. K., Celma M. L., Weissman S. M. The genome of simian virus 40. Science. 1978 May 5;200(4341):494–502. doi: 10.1126/science.205947. [DOI] [PubMed] [Google Scholar]
  17. Zweidler A. Resolution of histones by polyacrylamide gel electrophoresis in presence of nonionic detergents. Methods Cell Biol. 1978;17:223–233. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES