Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1977 Jan;21(1):215–224. doi: 10.1128/jvi.21.1.215-224.1977

Cell-free synthesis of polyoma virus capsid proteins VP1 and VP2.

T Wheeler, S T Bayley, R Harvey, L V Crawford, A E Smith
PMCID: PMC353807  PMID: 189065

Abstract

Polyadenylated RNA isolated from the cytoplasm of mouse 3T6 cells 28 h after infection with polyoma virus has been isolated and translated in vitro. Polyoma capsid proteins VP1 and VP2 have been identified in the cell-free product by polyacrylamide gel electrophoresis, specific immunoprecipitation, and tryptic peptide fingerprinting. Polyoma mRNA species have been isolated by preparative hybridization to purified viral DNA immobilized on cellulose nitrate filters and shown to code for both VP1 and VP2. These experiments establish conditions for the isolation of late polyoma mRNA and the cell-free synthesis of polyoma capsid proteins and indicate that the active mRNA species are at least partially virus coded.

Full text

PDF
223

Images in this article

217Image
on p.217
218Image
on p.218
219Image
on p.219
221Image
on p.221
222Image
on p.222

Selected References

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

  1. Crawford L. V., Gesteland R. F. Synthesis of polyoma proteins in vitro. J Mol Biol. 1973 Mar 15;74(4):627–634. doi: 10.1016/0022-2836(73)90053-3. [DOI] [PubMed] [Google Scholar]
  2. Crawford L. V. Proteins of polyoma virus and SV40. Br Med Bull. 1973 Sep;29(3):253–258. doi: 10.1093/oxfordjournals.bmb.a071016. [DOI] [PubMed] [Google Scholar]
  3. England J. M., Howett M. K., Tan K. B. Effect of hypertonic conditions on protein synthesis in cells productively infected with simian virus 40. J Virol. 1975 Nov;16(5):1101–1107. doi: 10.1128/jvi.16.5.1101-1107.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fey G., Hirt B. Fingerprints of polyoma virus proteins and mouse histones. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):235–241. doi: 10.1101/sqb.1974.039.01.030. [DOI] [PubMed] [Google Scholar]
  5. Frearson P. M., Crawford L. V. Polyoma virus basic proteins. J Gen Virol. 1972 Feb;14(2):141–155. doi: 10.1099/0022-1317-14-2-141. [DOI] [PubMed] [Google Scholar]
  6. Gibson W. Polyoma virus proteins: a description of the structural proteins of the virion based on polyacrylamide gel electrophoresis and peptide analysis. Virology. 1974 Dec;62(2):319–336. doi: 10.1016/0042-6822(74)90395-x. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Hewick R. M., Fried M., Waterfield M. D. Nonhistone virion proteins of polyoma: characterisation of the particle proteins by tryptic peptide analysis by use of ion-exchange columns. Virology. 1975 Aug;66(2):408–419. doi: 10.1016/0042-6822(75)90213-5. [DOI] [PubMed] [Google Scholar]
  9. 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]
  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., 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]
  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. Lazarides E., Weber K. Actin antibody: the specific visualization of actin filaments in non-muscle cells. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2268–2272. doi: 10.1073/pnas.71.6.2268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lindberg U., Persson T. Messenger RNA isolation with poly(U) agarose. Methods Enzymol. 1974;34:496–499. doi: 10.1016/s0076-6879(74)34061-x. [DOI] [PubMed] [Google Scholar]
  15. Miller L. K., Fried M. Construction of the genetic map of the polyoma genome. J Virol. 1976 Jun;18(3):824–832. doi: 10.1128/jvi.18.3.824-832.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Prives C. L., Aviv H., Gilboa E., Revel M., Winocour E. The cell-free translation of SV40 messenger RNA. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):309–316. doi: 10.1101/sqb.1974.039.01.041. [DOI] [PubMed] [Google Scholar]
  17. Prives C. L., Aviv H., Paterson B. M., Roberts B. E., Rozenblatt S., Revel M., Winocour E. Cell-free translation of messenger RNA of simian virus 40: synthesis of the major capsid protein. Proc Natl Acad Sci U S A. 1974 Feb;71(2):302–306. doi: 10.1073/pnas.71.2.302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roberts B. E., Paterson B. M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330–2334. doi: 10.1073/pnas.70.8.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  20. Smith A. E., Kamen R., Mangel W. F., Shure H., Wheeler T. Location of the sequences coding for capsid proteins VP1 and VP2 on polyoma virus DNA. Cell. 1976 Nov;9(3):481–487. doi: 10.1016/0092-8674(76)90093-3. [DOI] [PubMed] [Google Scholar]
  21. Smith A. E., Wheeler T., Glanville N., Käriäinen Translation of Semliki-Forest-virus 42-S RNA in a mouse cell free system to give virus-coat proteins. Eur J Biochem. 1974 Nov 1;49(1):101–110. doi: 10.1111/j.1432-1033.1974.tb03815.x. [DOI] [PubMed] [Google Scholar]
  22. TODARO G. J., GREEN H. Quantitative studies of the growth of mouse embryo cells in culture and their development into established lines. J Cell Biol. 1963 May;17:299–313. doi: 10.1083/jcb.17.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Walter G., Deppert W. Intermolecular disulfide bonds: an important structural feature of the polyoma virus capsid. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):255–257. doi: 10.1101/sqb.1974.039.01.033. [DOI] [PubMed] [Google Scholar]
  24. Westphal H., Eron L., Ferdinand F. J., Callahan R., Lai S. P. Analysis of adenovirus type 2 gene functions by cell-free translation of viral messenger RNA. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):575–579. doi: 10.1101/sqb.1974.039.01.071. [DOI] [PubMed] [Google Scholar]

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

RESOURCES