Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1987 Feb;61(2):509–515. doi: 10.1128/jvi.61.2.509-515.1987

Differences in biological activity and structural protein VP1 phosphorylation of polyomavirus progeny resulting from infection of primary mouse kidney and primary mouse embryo cell cultures.

J W Ludlow, R A Consigli
PMCID: PMC253975  PMID: 3027379

Abstract

Both primary mouse kidney and primary mouse embryo cells in culture were used for polyomavirus progeny production. Examination of polyomavirus virion structural integrity revealed that mouse embryo cell progeny contained a threefold greater population of unstable particles when compared with mouse kidney cell progeny. Differences in biological activity between these two progeny virion types were also shown. Mouse kidney cell progeny compared with mouse embryo cell progeny exhibited a 10-fold greater ability to agglutinate guinea pig erythrocytes, a 3-fold lower ability to become internalized into monopinocytotic vesicles, and a 2-fold lower ability to initiate a productive infection based on positive nuclear immunofluorescence when mouse embryo host cell cultures were used. The mouse kidney progeny were also found to bind to host cells less specifically than the mouse embryo cell progeny. When these two progeny virion types were labeled in vivo with 32P and subjected to isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophroesis in the second dimension, differences in the phosphorylation pattern of the major virus-encoded structural protein VP1 species were observed. It was revealed that species D and E of mouse kidney cell progeny were phosphorylated to the same degree, while mouse embryo cell progeny species E and F were phosphorylated equally. These data suggest that the host cells play a role in modulating the biological activity of the virus by affecting the degree and site-specific phosphorylation of the major capsid protein VP1 which may influence the recognition of virus attachment proteins for specific cellular receptors.

Full text

PDF
509

Images in this article

511Image
on p.511

Selected References

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

  1. Anders D. G., Consigli R. A. Chemical cleavage of polyomavirus major structural protein VP1: identification of cleavage products and evidence that the receptor moiety resides in the carboxy-terminal region. J Virol. 1983 Oct;48(1):197–205. doi: 10.1128/jvi.48.1.197-205.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anders D. G., Consigli R. A. Comparison of nonphosphorylated and phosphorylated species of polyomavirus major capsid protein VP1 and identification of the major phosphorylation region. J Virol. 1983 Oct;48(1):206–217. doi: 10.1128/jvi.48.1.206-217.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolen J. B., Anders D. G., Trempy J., Consigli R. A. Differences in the subpopulations of the structural proteins of polyoma virions and capsids: biological functions of the multiple VP1 species. J Virol. 1981 Jan;37(1):80–91. doi: 10.1128/jvi.37.1.80-91.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolen J. B., Consigli R. A. Differential adsorption of polyoma virions and capsids to mouse kidney cells and guinea pig erythrocytes. J Virol. 1979 Nov;32(2):679–683. doi: 10.1128/jvi.32.2.679-683.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bolen J. B., Consigli R. A. Separation of neutralizing and hemagglutination-inhibiting antibody activities and specificity of antisera to sodium dodecyl sulfate-derived polypeptides of polyoma virions. J Virol. 1980 Apr;34(1):119–129. doi: 10.1128/jvi.34.1.119-129.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Brunk C. F., Leick V. Rapid equilibrium isopycnic CsC1 gradients. Biochim Biophys Acta. 1969 Mar 18;179(1):136–144. doi: 10.1016/0005-2787(69)90129-4. [DOI] [PubMed] [Google Scholar]
  9. CRAWFORD L. V., CRAWFORD E. M., WATSONDH The physical characteristics of polyoma virus. I. Two types of particle. Virology. 1962 Oct;18:170–176. doi: 10.1016/0042-6822(62)90002-8. [DOI] [PubMed] [Google Scholar]
  10. CRAWFORD L. V. The adsorption of polyoma virus. Virology. 1962 Oct;18:177–181. doi: 10.1016/0042-6822(62)90003-x. [DOI] [PubMed] [Google Scholar]
  11. Consigli R. A., Minocha H. C., Abo-Ahmed H. The effect of 5-fluoro-deoxyuridine on the replication of viral DNA and synthesis of polyoma capsid protein. J Gen Virol. 1968 May;2(3):437–441. doi: 10.1099/0022-1317-2-3-437. [DOI] [PubMed] [Google Scholar]
  12. Etchison D., Walter G. Subunit interactions in polyoma virus structure. Virology. 1977 Apr;77(2):783–796. doi: 10.1016/0042-6822(77)90499-8. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. Garcea R. L., Ballmer-Hofer K., Benjamin T. L. Virion assembly defect of polyomavirus hr-t mutants: underphosphorylation of major capsid protein VP1 before viral DNA encapsidation. J Virol. 1985 May;54(2):311–316. doi: 10.1128/jvi.54.2.311-316.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Griffith G. R., Consigli R. A. Cross-linking of a polyomavirus attachment protein to its mouse kidney cell receptor. J Virol. 1986 Jun;58(3):773–781. doi: 10.1128/jvi.58.3.773-781.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Griffith G. R., Consigli R. A. Isolation and characterization of monopinocytotic vesicles containing polyomavirus from the cytoplasm of infected mouse kidney cells. J Virol. 1984 Apr;50(1):77–85. doi: 10.1128/jvi.50.1.77-85.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kistner O., Müller H., Becht H., Scholtissek C. Phosphopeptide fingerprints of nucleoproteins of various influenza A virus strains grown in different host cells. J Gen Virol. 1985 Mar;66(Pt 3):465–472. doi: 10.1099/0022-1317-66-3-465. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. MORI R., SCHIEBLE J. H., ACKERMANN W. W. Reaction of polyoma and influenza viruses with receptors of erythrocytes and host-cells. Proc Soc Exp Biol Med. 1962 Mar;109:685–690. doi: 10.3181/00379727-109-27307. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. 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]
  25. McMillen J., Consigli R. A. In vitro radioisotopic labeling of proteins associated with purified polyoma virions. J Virol. 1974 Dec;14(6):1627–1629. doi: 10.1128/jvi.14.6.1627-1629.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Moore B. R., Free S. J. Protein modification and its biological role. Int J Biochem. 1985;17(3):283–289. doi: 10.1016/0020-711x(85)90202-2. [DOI] [PubMed] [Google Scholar]
  27. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  28. O'Farrell P. Z., Goodman H. M. Resolution of simian virus 40 proteins in whole cell extracts by two-dimensional electrophoresis: heterogeneity of the major capsid protein. Cell. 1976 Oct;9(2):289–298. doi: 10.1016/0092-8674(76)90119-7. [DOI] [PubMed] [Google Scholar]
  29. Rayment I., Baker T. S., Caspar D. L., Murakami W. T. Polyoma virus capsid structure at 22.5 A resolution. Nature. 1982 Jan 14;295(5845):110–115. doi: 10.1038/295110a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roblin R., Härle E., Dulbecco R. Polyoma virus proteins. 1. Multiple virion components. Virology. 1971 Sep;45(3):555–566. doi: 10.1016/0042-6822(71)90171-1. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. WINOCOUR E. Purification of polyoma virus. Virology. 1963 Feb;19:158–168. doi: 10.1016/0042-6822(63)90005-9. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Winston V. D., Bolen J. B., Consigli R. A. Isolation and characterization of polyoma uncoating intermediates from the nuclei of infected mouse cells. J Virol. 1980 Mar;33(3):1173–1181. doi: 10.1128/jvi.33.3.1173-1181.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Yuen L. K., Consigli R. A. Generation of capsids from unstable polyoma virions. J Virol. 1983 Sep;47(3):620–625. doi: 10.1128/jvi.47.3.620-625.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yuen L. K., Consigli R. A. Identification and protein analysis of polyomavirus assembly intermediates from infected primary mouse embryo cells. Virology. 1985 Jul 15;144(1):127–138. doi: 10.1016/0042-6822(85)90311-3. [DOI] [PubMed] [Google Scholar]

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

RESOURCES