Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1990 Sep;64(9):4414–4420. doi: 10.1128/jvi.64.9.4414-4420.1990

Myristylated polyomavirus VP2: role in the life cycle of the virus.

N Krauzewicz 1, C H Streuli 1, N Stuart-Smith 1, M D Jones 1, S Wallace 1, B E Griffin 1
PMCID: PMC247910  PMID: 2166822

Abstract

The double-stranded genome of the small DNA tumor virus, polyomavirus, is enclosed in a capsid composed of a major protein, VP1, which associates as pentameric capsomeres into an icosahedral structure, and two minor proteins, VP2 and VP3, whose functions and positions within the structure are unknown. The N-terminal glycine of the VP2 coat protein has been shown to be cotranslationally acylated with myristic acid. To study the function of this modification and the role of VP2 in the life cycle of polyomavirus, the N-terminal glycine, critical to the myristylation consensus sequence, has been altered to a glutamic acid or a valine residue by site-directed oligonucleotide mutagenesis. The glycine----glutamic acid mutant DNA has been further studied. When transfected into cells permissive for the polyomavirus full lytic life cycle, this mutant DNA replicated at levels comparable to those of wild-type viral DNA, and small amounts of nonrevertant (mutant) virus could be harvested from the cultures. The virus particles viewed by electron microscopy appeared slightly distorted, but the ratio of full to empty particles was similar to that produced in a wild-type viral infection. Mutant virus was capable of reinfecting permissive cells but with a considerably reduced efficiency.

Full text

PDF
4415

Images in this article

4417Image
on p.4417
4418Image
on p.4418

Selected References

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

  1. Birnboim H. C. A rapid alkaline extraction method for the isolation of plasmid DNA. Methods Enzymol. 1983;100:243–255. doi: 10.1016/0076-6879(83)00059-2. [DOI] [PubMed] [Google Scholar]
  2. Clayson E. T., Brando L. V., Compans R. W. Release of simian virus 40 virions from epithelial cells is polarized and occurs without cell lysis. J Virol. 1989 May;63(5):2278–2288. doi: 10.1128/jvi.63.5.2278-2288.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cole C. N., Landers T., Goff S. P., Manteuil-Brutlag S., Berg P. Physical and genetic characterization of deletion mutants of simian virus 40 constructed in vitro. J Virol. 1977 Oct;24(1):277–294. doi: 10.1128/jvi.24.1.277-294.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dales S. Early events in cell-animal virus interactions. Bacteriol Rev. 1973 Jun;37(2):103–135. doi: 10.1128/br.37.2.103-135.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dilworth S. M., Griffin B. E. Monoclonal antibodies against polyoma virus tumor antigens. Proc Natl Acad Sci U S A. 1982 Feb;79(4):1059–1063. doi: 10.1073/pnas.79.4.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duckworth M. L., Gait M. J., Goelet P., Hong G. F., Singh M., Titmas R. C. Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route. Nucleic Acids Res. 1981 Apr 10;9(7):1691–1706. doi: 10.1093/nar/9.7.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emini E. A., Jameson B. A., Wimmer E. Priming for and induction of anti-poliovirus neutralizing antibodies by synthetic peptides. Nature. 1983 Aug 25;304(5928):699–703. doi: 10.1038/304699a0. [DOI] [PubMed] [Google Scholar]
  8. Franke C. A., Reynolds P. L., Hruby D. E. Fatty acid acylation of vaccinia virus proteins. J Virol. 1989 Oct;63(10):4285–4291. doi: 10.1128/jvi.63.10.4285-4291.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Griffin B. E., Fried M. Amplification of a specific region of the polyoma virus genome. Nature. 1975 Jul 17;256(5514):175–179. doi: 10.1038/256175a0. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Griffith G. R., Marriott S. J., Rintoul D. A., Consigli R. A. Early events in polyomavirus infection: fusion of monopinocytotic vesicles containing virions with mouse kidney cell nuclei. Virus Res. 1988 Apr;10(1):41–51. doi: 10.1016/0168-1702(88)90056-1. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. Marc D., Drugeon G., Haenni A. L., Girard M., van der Werf S. Role of myristoylation of poliovirus capsid protein VP4 as determined by site-directed mutagenesis of its N-terminal sequence. EMBO J. 1989 Sep;8(9):2661–2668. doi: 10.1002/j.1460-2075.1989.tb08406.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Marriott S. J., Griffith G. R., Consigli R. A. Octyl-beta-D-glucopyranoside extracts polyomavirus receptor moieties from the surfaces of mouse kidney cells. J Virol. 1987 Feb;61(2):375–382. doi: 10.1128/jvi.61.2.375-382.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. NEGRONI G., DOURMASHKIN R., CHESTERMAN F. C. A "polyoma" virus derived from a mouse leukaemia. Br Med J. 1959 Dec 19;2(5163):1359–1360. doi: 10.1136/bmj.2.5163.1359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Payne L. G., Norrby E. Adsorption and penetration of enveloped and naked vaccinia virus particles. J Virol. 1978 Jul;27(1):19–27. doi: 10.1128/jvi.27.1.19-27.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pevear D. C., Fancher M. J., Felock P. J., Rossmann M. G., Miller M. S., Diana G., Treasurywala A. M., McKinlay M. A., Dutko F. J. Conformational change in the floor of the human rhinovirus canyon blocks adsorption to HeLa cell receptors. J Virol. 1989 May;63(5):2002–2007. doi: 10.1128/jvi.63.5.2002-2007.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stamatos N. M., Chakrabarti S., Moss B., Hare J. D. Expression of polyomavirus virion proteins by a vaccinia virus vector: association of VP1 and VP2 with the nuclear framework. J Virol. 1987 Feb;61(2):516–525. doi: 10.1128/jvi.61.2.516-525.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Strauss M., Streuli C. H., Griffin B. E. Efficient oligodeoxyribonucleotide-directed deletion mutagenesis using pEMBL vectors: removal of early region introns from polyoma virus mutants. Gene. 1986;49(3):331–340. doi: 10.1016/0378-1119(86)90369-0. [DOI] [PubMed] [Google Scholar]
  22. Streuli C. H., Griffin B. E. Myristic acid is coupled to a structural protein of polyoma virus and SV40. Nature. 1987 Apr 9;326(6113):619–622. doi: 10.1038/326619a0. [DOI] [PubMed] [Google Scholar]
  23. Towler D. A., Gordon J. I., Adams S. P., Glaser L. The biology and enzymology of eukaryotic protein acylation. Annu Rev Biochem. 1988;57:69–99. doi: 10.1146/annurev.bi.57.070188.000441. [DOI] [PubMed] [Google Scholar]
  24. Twigg A. J., Sherratt D. Trans-complementable copy-number mutants of plasmid ColE1. Nature. 1980 Jan 10;283(5743):216–218. doi: 10.1038/283216a0. [DOI] [PubMed] [Google Scholar]
  25. Tycko B., Maxfield F. R. Rapid acidification of endocytic vesicles containing alpha 2-macroglobulin. Cell. 1982 Mar;28(3):643–651. doi: 10.1016/0092-8674(82)90219-7. [DOI] [PubMed] [Google Scholar]
  26. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis: a simple method using two oligonucleotide primers and a single-stranded DNA template. Methods Enzymol. 1987;154:329–350. doi: 10.1016/0076-6879(87)54083-6. [DOI] [PubMed] [Google Scholar]

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

RESOURCES