1887

Abstract

A mutational and genetic analysis of the poliovirus protein 3A has led to the identification of a single amino acid mutant virus with a restrictive phenotype to form plaques in Vero cells. This mutant (I46T 3A) can be grown and amplified in HeLa cells, where virus replication takes place at wild-type levels. However, Vero cells infected with this virus cannot complete the growth cycle. I46T 3A virus has a defect in the ability to kill Vero cells, as estimated by FACS analysis of propidium iodide uptake by dead cells. Since these defects are observed under conditions where no abnormalities in the rate of synthesis and processing of the mutant polyprotein occur, the inability to induce the cytopathic effect in infected Vero cells denotes the existence of a defect in the activity of 3A, but not the level of expression of the viral genome. As a consequence of this impaired capability to generate the cytopathic effect, I46T 3A mutant viruses cannot be titrated by plaque assay in Vero cells. Only revertant viruses with the wild-type sequence arise and form lysis plaques in Vero cells. Our results suggest a role for the 3A protein (or a precursor thereof) in the virus- induced cytopathic effect. The mutant virus characterized in this work may be a useful tool to understand how poliovirus kills infected cells and carries out the final step of its life-cycle, the release of virus progeny.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-79-8-1911
1998-08-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/79/8/9714239.html?itemId=/content/journal/jgv/10.1099/0022-1317-79-8-1911&mimeType=html&fmt=ahah

References

  1. Bienz K., Egger D., Rasser Y., Bossart W. 1983; Intracellular distribution of poliovirus proteins and the induction of virus-specific cytoplasmic structures. Virology 131:39–48
    [Google Scholar]
  2. Bowen I. D. 1993; Apoptosis or programmed cell death?. Cell Biology International 17:365–380
    [Google Scholar]
  3. Carrasco L. 1977; The inhibition of cell functions after viral infection. A proposed general mechanism. FEBS Letters 76:11–15
    [Google Scholar]
  4. Carrasco L. 1987 Mechanisms of Viral Toxicity in Animal Cells pp. 1–3 Raton: CRC Press;
    [Google Scholar]
  5. Carrasco L., Otero M. J., Castrillo J. L. 1989; Modification of membrane permeability by animal viruses. Pharmacology & Therapeutics 40171–212
    [Google Scholar]
  6. Carrasco L., Perez L., Irurzun A., Lama J., Martinez-Abarca F., Rodrigez P., Guinea R., Castrillo J. L., Sanz M. A., Ayala M. J. 1993; Modification of membrane permeability by animal viruses. pp. 283–305 In Regulation of Gene Expression in Animal Viruses Carrasco L., Sonenberg N., Wimmer E. Edited by London & New York: Plenum Press;
    [Google Scholar]
  7. Castrillo J. L., López-Rivas A., Carrasco L. 1987; Effects of extracellular cations on translation in poliovirus-infected cells. Journal of General Virology 68:325–333
    [Google Scholar]
  8. Cho M. W., Teterina N., Egger D., Bienz K., Ehrenfeld E. 1994; Membrane rearrangement and vesicle induction by recombinant poliovirus 2C and 2BC in human cells. Virology 202:129–145
    [Google Scholar]
  9. Clark M. E., Dasgupta A. 1990; A transcriptionally active form of TFIIIC is modified in poliovirus-infected HeLa cells. Molecular and Cellular Biology 10:5106–5113
    [Google Scholar]
  10. Clark M. E., Hämmerle T., Wimmer E., Dasgupta A. 1991; Poliovirus proteinase 3C converts an active form of transcription factor IIIC to an inactive form, a mechanism for inhibition of host cell polymerase III transcription by poliovirus. EMBO Journal 10:2941–2947
    [Google Scholar]
  11. Contreras A., Carrasco L. 1979; Selective inhibition of protein synthesis in virus-infected mammalian cells. Journal of Virology 29:114–122
    [Google Scholar]
  12. Esolen L. M., Park S. W., Hardwick M., Griffin D. E. 1995; Apoptosis as a cause of death in measles virus-infected cells. Journal of Virology 39:3955–3958
    [Google Scholar]
  13. Etchison D., Milburn S. C., Edery I., Sonenberg N., Hershey J. W. 1982; Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220,000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. Journal of Biological Chemistry 257:14806–14810
    [Google Scholar]
  14. Giachetti C., Hwang S.-S., Semler B. L. 1992; cis-Acting lesions targeted to the hydrophobic domain of a poliovirus membrane protein involved in RNA replication. Journal of Virology 66:6045–6057
    [Google Scholar]
  15. Gray J. W., Coffino P. 1979; Cell cycle analysis by flow cytometry. Methods in Enzymology 58:233–248
    [Google Scholar]
  16. Harris K. S., Reddigari S. R., Nicklin M. J. H., Hämmerle T., Wimmer E. 1992; Purification and characterization of poliovirus polypeptide 3CD, a proteinase and a precursor for RNA polymerase. Journal of Virology 66:7481–7489
    [Google Scholar]
  17. Hinshaw V. S., Olsen C., Dybdahl-Sissoko N., Evans D. 1994; Apoptosis, a mechanism of cell killing by influenza A and B viruses. Journal of Virology 68:3667–3673
    [Google Scholar]
  18. Holland J. J. 1964; Inhibition of host cell macromolecular synthesis by high multiplicities of poliovirus under conditions preventing virus synthesis. Journal of Molecular Biology 8:574–581
    [Google Scholar]
  19. Irurzun A., Arroyo J., Alvarez A., Carrasco L. 1995; Enhanced intracellular calcium concentration during poliovirus infection. Journal of Virology 69:5142–5146
    [Google Scholar]
  20. Jeurissen S. H. M., Wagenaar F., Pol J. M. A., van der Eb A., Noteborn M. H. M. 1992; Chicken anemia virus causes apoptosis of thymocytes after in vivo infection and of cell lines after in vitro infection. Journal of Virology 66:7383–7388
    [Google Scholar]
  21. Koch F., Koch G. 1985 The Molecular Biology of Poliovirus Wien: Springer Verlag;
    [Google Scholar]
  22. Krausslich H. G., Nicklin M. J., Toyoda H., Etchison D., Wimmer E. 1987; Poliovirus proteinase 2A induces cleavage of eucaryotic initiation factor 4F polypeptide p220. Journal of Virology 61:2711–2718
    [Google Scholar]
  23. Lama J., Carrasco L. 1992; Expression of poliovirus nonstructural proteins in Escherichia coli cells. Modification of membrane permeability induced by 2B and 3A. Journal of Biological Chemistry 267:15932–15937
    [Google Scholar]
  24. Lama J., Carrasco L. 1995; Mutations in the hydrophobic domain of poliovirus protein 3AB abrogate its permeabilizing activity. FEBS Letters 367:5–11
    [Google Scholar]
  25. Lama J., Carrasco L. 1996; Screening for membrane-permeabilizing mutants of the poliovirus protein 3AB. Journal of General Virology 77:2109–2119
    [Google Scholar]
  26. Lama J., Paul A. V., Harris K. S., Wimmer E. 1994; Properties of purified recombinant poliovirus protein 3AB as substrate for viral proteinases and as a cofactor for RNA polymerase 3Dpol. Journal of Biological Chemistry 269:66–70
    [Google Scholar]
  27. Lama J., Sanz M. A., Rodriguez P. L. 1995; A role for 3AB protein in poliovirus genome replication. Journal of Biological Chemistry 270:14430–14438
    [Google Scholar]
  28. Laurent-Crawford A. G., Krust B., Rivière Y., Desgranges C., Muller S., Kieny M. P., Dauguet C., Hovanessian A. G. 1993; Membrane expression of HIV envelope glycoproteins triggers apoptosis in CD4 cells. AIDS Research and Human Retroviruses 9:761–773
    [Google Scholar]
  29. Levine B., Huang Q., Isaacs J. T., Reed J. C., Griffin D. E., Hardwick J. M. 1993; Conversion of lytic to persistent alphavirus infection by the bcl-2 cellular oncogene. Nature 361:739–742
    [Google Scholar]
  30. López-Rivas A., Castrillo J. L., Carrasco L. 1987; Cation content in poliovirus-infected HeLa cells. Journal of General Virology 68:335–342
    [Google Scholar]
  31. Michelangeli F., Ruiz M. C., del Castillo J. R., Ludert J. E., Liprandi F. 1991; Effect of rotavirus infection on intracellular calcium homeostasis in cultured cells. Virology 181:520–527
    [Google Scholar]
  32. Molla A., Paul A. V., Wimmer E. 1991; Cell-free, de novo synthesis of poliovirus. Science 254:1647–1651
    [Google Scholar]
  33. Molla A., Harris K., Paul A. V., Shin S., Mugavero J., Wimmer E. 1994; Stimulation of poliovirus proteinase 3Cpro related proteolysis by the genome linked protein VPg and its precursor 3AB. Journal of Biological Chemistry 269:27015–27020
    [Google Scholar]
  34. Morace G., Gaus-Muller V., Beneduce F., Pisani G. 1993; Mutations in the 3A genomic region of two cytopathic strains of hepatitis A virus isolated in Italy. Virus Research 28:187–194
    [Google Scholar]
  35. Ortin J., Najera R., Lopez C., Davila E., Domingo E. 1980; Genetic variability of Hong Kong (H3N2) influenza viruses. Spontaneous mutations and their location in the viral genome. Gene 11:319–331
    [Google Scholar]
  36. Pelletier J., Sonenberg N. 1988; Internal initiation oftranslation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature 334:320–325
    [Google Scholar]
  37. Semler B. L., Anderson C. W., Hanecak R., Dorner L. F., Wimmer E. 1982; A membrane-associated precursor to poliovirus VPg identified by immunoprecipitation with antibodies directed against a synthetic heptapeptide. Cell 28:405–412
    [Google Scholar]
  38. Tolskaya A., Romanova L., Kolesnikova M. S., Ivannikova T. A., Smirnova E. A., Raikhlin N. T., Agol V. I. 1995; Apoptosis-inducing and apoptosis-preventing functions of poliovirus. Journal of Virology 69:1181–1189
    [Google Scholar]
  39. van der Werf S., Bradley J., Wimmer E., Studier F., Dunn J. J. 1986; Synthesis of infectious poliovirus RNA by purified T7 RNA polymerase. Proceedings of the National Academy of Sciences, USA 83:2330–2334
    [Google Scholar]
  40. van Kuppeveld F. J., Galama J. M., Zoll J., van der Hurk P. J., Melchers W. J. 1996; Coxsackie B3 virus protein 2B contains a cationic amphipathic helix that is required for viral RNA replication. Journal of Virology 70:3876–3886
    [Google Scholar]
  41. Wimmer E., Hellen C. U. T., Cao X. 1993; Genetics of poliovirus. Annual Review of Genetics 27:353–436
    [Google Scholar]
  42. Young R. 1992; Bacteriophage lysis: mechanism and regulation. Microbiological Reviews 56:430–481
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-79-8-1911
Loading
/content/journal/jgv/10.1099/0022-1317-79-8-1911
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error