Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1994 May;68(5):3386–3390. doi: 10.1128/jvi.68.5.3386-3390.1994

Purification of recombinant budgerigar fledgling disease virus VP1 capsid protein and its ability for in vitro capsid assembly.

R E Rodgers 1, D Chang 1, X Cai 1, R A Consigli 1
PMCID: PMC236831  PMID: 8151798

Abstract

A recombinant system for the major capsid VP1 protein of budgerigar fledgling disease virus has been established. The VP1 gene was inserted into a truncated form of the pFlag-1 vector and expressed in Escherichia coli. The budgerigar fledgling disease virus VP1 protein was purified to near homogeneity by immunoaffinity chromatography. Fractions containing highly purified VP1 were pooled and found to constitute 3.3% of the original E. coli-expressed VP1 protein. Electron microscopy revealed that the VP1 protein was isolated as pentameric capsomeres. Electron microscopy also revealed that capsid-like particles were formed in vitro from purified VP1 capsomeres with the addition of Ca2+ ions and the removal of chelating and reducing agents.

Full text

PDF
3386

Images in this article

3387Image
on p.3387
3388Image
on p.3388
3389Image
on p.3389

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. Bernier G., Morin M., Marsolais G. A generalized inclusion body disease in the budgerigar (Melopsittacus undulatus) caused by a papovavirus-like agent. Avian Dis. 1981 Oct-Dec;25(4):1083–1092. [PubMed] [Google Scholar]
  4. 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]
  5. Bozeman L. H., Davis R. B., Gaudry D., Lukert P. D., Fletcher O. J., Dykstra M. J. Characterization of a papovavirus isolated from fledgling budgerigars. Avian Dis. 1981 Oct-Dec;25(4):972–980. [PubMed] [Google Scholar]
  6. Brady J. N., Consigli R. A. Chromatographic separation of the polyoma virus proteins and renaturation of the isolated VP1 major capsid protein. J Virol. 1978 Aug;27(2):436–442. doi: 10.1128/jvi.27.2.436-442.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brady J. N., Kendall J. D., Consigli R. A. In vitro reassembly of infectious polyoma virions. J Virol. 1979 Nov;32(2):640–647. doi: 10.1128/jvi.32.2.640-647.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Chang D., Cai X., Consigli R. A. Characterization of the DNA binding properties of polyomavirus capsid protein. J Virol. 1993 Oct;67(10):6327–6331. doi: 10.1128/jvi.67.10.6327-6331.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chang D., Haynes J. I., 2nd, Brady J. N., Consigli R. A. The use of additive and subtractive approaches to examine the nuclear localization sequence of the polyomavirus major capsid protein VP1. Virology. 1992 Aug;189(2):821–827. doi: 10.1016/0042-6822(92)90615-v. [DOI] [PubMed] [Google Scholar]
  12. Christiansen G., Landers T., Griffith J., Berg P. Characterization of components released by alkali disruption of simian virus 40. J Virol. 1977 Mar;21(3):1079–1084. doi: 10.1128/jvi.21.3.1079-1084.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Davis R. B., Bozeman L. H., Gaudry D., Fletcher O. J., Lukert P. D., Dykstra M. J. A viral disease of fledgling budgerigars. Avian Dis. 1981 Jan-Mar;25(1):179–183. [PubMed] [Google Scholar]
  14. Dykstra M. J., Dykstra C. C., Lukert P. D., Bozeman L. H. Investigations of budgerigar fledgling disease virus. Am J Vet Res. 1984 Sep;45(9):1883–1887. [PubMed] [Google Scholar]
  15. Fattaey A. R., Consigli R. A. Synthesis, posttranslational modifications, and nuclear transport of polyomavirus major capsid protein VP1. J Virol. 1989 Jul;63(7):3168–3175. doi: 10.1128/jvi.63.7.3168-3175.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fattaey A., Lenz L., Consigli R. A. Production and characterization of monoclonal antibodies to budgerigar fledgling disease virus major capsid protein VP. Avian Dis. 1992 Jul-Sep;36(3):543–553. [PubMed] [Google Scholar]
  17. 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]
  18. Hagensee M. E., Yaegashi N., Galloway D. A. Self-assembly of human papillomavirus type 1 capsids by expression of the L1 protein alone or by coexpression of the L1 and L2 capsid proteins. J Virol. 1993 Jan;67(1):315–322. doi: 10.1128/jvi.67.1.315-322.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Haynes J. I., 2nd, Chang D., Consigli R. A. Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly. J Virol. 1993 May;67(5):2486–2495. doi: 10.1128/jvi.67.5.2486-2495.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Haynes J. I., 2nd, Consigli R. A. Phosphorylation of the budgerigar fledgling disease virus major capsid protein VP1. J Virol. 1992 Jul;66(7):4551–4555. doi: 10.1128/jvi.66.7.4551-4555.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hunter B., Gagnon A., Onderka D., Goltz J., Holmes B. Viral hepatitis in budgerigars in southern Ontario. Can Vet J. 1979 Jun;20(6):176–176. [PMC free article] [PubMed] [Google Scholar]
  22. Kaleta E. F., Herbst W., Kaup F. J., Jank-Ladwig R., Marschall H. J., Drommer W., Krautwald M. Untersuchungen zur Virusätiologie einer mit Hepatitis und Befiederungsstörungen einhergehender Krankheit bei Wellesittich-Nestlingen (Melopsittacus undulatus). Zentralbl Veterinarmed B. 1984 Apr;31(3):219–224. [PubMed] [Google Scholar]
  23. Kirnbauer R., Taub J., Greenstone H., Roden R., Dürst M., Gissmann L., Lowy D. R., Schiller J. T. Efficient self-assembly of human papillomavirus type 16 L1 and L1-L2 into virus-like particles. J Virol. 1993 Dec;67(12):6929–6936. doi: 10.1128/jvi.67.12.6929-6936.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lawler J., Hynes R. O. The structure of human thrombospondin, an adhesive glycoprotein with multiple calcium-binding sites and homologies with several different proteins. J Cell Biol. 1986 Nov;103(5):1635–1648. doi: 10.1083/jcb.103.5.1635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lehn H., Müller H. Cloning and characterization of budgerigar fledgling disease virus, an avian polyomavirus. Virology. 1986 Jun;151(2):362–370. doi: 10.1016/0042-6822(86)90056-5. [DOI] [PubMed] [Google Scholar]
  26. Ludlow J. W., Consigli R. A. 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 Virol. 1987 Feb;61(2):509–515. doi: 10.1128/jvi.61.2.509-515.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ludlow J. W., Consigli R. A. Hydroxyproline in the major capsid protein VP1 of polyomavirus. J Virol. 1989 Jun;63(6):2881–2884. doi: 10.1128/jvi.63.6.2881-2884.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ludlow J. W., Consigli R. A. Localization of calcium on the polyomavirus VP1 capsid protein. J Virol. 1987 Sep;61(9):2934–2937. doi: 10.1128/jvi.61.9.2934-2937.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Ludlow J. W., Consigli R. A. Polyomavirus major capsid protein VP1 is modified by tyrosine sulfuration. J Virol. 1987 May;61(5):1708–1711. doi: 10.1128/jvi.61.5.1708-1711.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. Minocha H. C., Consigli R. A., Eisenstark A. Concentration and purification of Newcastle disease virus. Am J Vet Res. 1968 Apr;29(4):877–882. [PubMed] [Google Scholar]
  32. Montross L., Watkins S., Moreland R. B., Mamon H., Caspar D. L., Garcea R. L. Nuclear assembly of polyomavirus capsids in insect cells expressing the major capsid protein VP1. J Virol. 1991 Sep;65(9):4991–4998. doi: 10.1128/jvi.65.9.4991-4998.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Moreland R. B., Garcea R. L. Characterization of a nuclear localization sequence in the polyomavirus capsid protein VP1. Virology. 1991 Nov;185(1):513–518. doi: 10.1016/0042-6822(91)90811-o. [DOI] [PubMed] [Google Scholar]
  34. Moreland R. B., Montross L., Garcea R. L. Characterization of the DNA-binding properties of the polyomavirus capsid protein VP1. J Virol. 1991 Mar;65(3):1168–1176. doi: 10.1128/jvi.65.3.1168-1176.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pipas J. M. Common and unique features of T antigens encoded by the polyomavirus group. J Virol. 1992 Jul;66(7):3979–3985. doi: 10.1128/jvi.66.7.3979-3985.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ponder B. A., Robbins A. K., Crawford L. V. Phophorylation of polyoma and SV40 virus proteins. J Gen Virol. 1977 Oct;37(1):75–83. doi: 10.1099/0022-1317-37-1-75. [DOI] [PubMed] [Google Scholar]
  37. Rose R. C., Bonnez W., Reichman R. C., Garcea R. L. Expression of human papillomavirus type 11 L1 protein in insect cells: in vivo and in vitro assembly of viruslike particles. J Virol. 1993 Apr;67(4):1936–1944. doi: 10.1128/jvi.67.4.1936-1944.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rott O., Kröger M., Müller H., Hobom G. The genome of budgerigar fledgling disease virus, an avian polyomavirus. Virology. 1988 Jul;165(1):74–86. doi: 10.1016/0042-6822(88)90660-5. [DOI] [PubMed] [Google Scholar]
  39. Salunke D. M., Caspar D. L., Garcea R. L. Polymorphism in the assembly of polyomavirus capsid protein VP1. Biophys J. 1989 Nov;56(5):887–900. doi: 10.1016/S0006-3495(89)82735-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Salunke D. M., Caspar D. L., Garcea R. L. Self-assembly of purified polyomavirus capsid protein VP1. Cell. 1986 Sep 12;46(6):895–904. doi: 10.1016/0092-8674(86)90071-1. [DOI] [PubMed] [Google Scholar]
  41. Schmidt M., Müller H., Schmidt M. F., Rott R. Myristoylation of budgerigar fledgling disease virus capsid protein VP2. J Virol. 1989 Jan;63(1):429–431. doi: 10.1128/jvi.63.1.429-431.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Strynadka N. C., James M. N. Crystal structures of the helix-loop-helix calcium-binding proteins. Annu Rev Biochem. 1989;58:951–998. doi: 10.1146/annurev.bi.58.070189.004511. [DOI] [PubMed] [Google Scholar]
  43. Yuen L. K., Consigli R. A. Improved infectivity of reassembled polyoma virus. J Virol. 1982 Jul;43(1):337–341. doi: 10.1128/jvi.43.1.337-341.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES