1887

Abstract

Prion replication involves conversion of the normal, host-encoded prion protein PrP, which is a sialoglycoprotein bound to the plasma membrane by a glycophosphatidylinositol anchor, into a pathogenic isoform, PrP. In earlier studies, tunicamycin prevented glycosylation of PrP in scrapie-infected mouse neuroblastoma (ScN2a) cells but it was still expressed on the cell surface and converted into PrP; mutation of PrP at glycosylation consensus sites (T182A, T198A) produced low steady-state levels of PrP that were insufficient to propagate prions in transgenic mice. By mutating asparagines to glutamines at the consensus sites, we obtained expression of unglycosylated, epitope-tagged MHM2PrP(N180Q,N196Q), which was converted into PrP in ScN2a cells. Cultures of uninfected neuroblastoma (N2a) cells transiently expressing mutated PrP were exposed to brain homogenates prepared from mice infected with the RML, Me7 or 301V prion strains. In each case, mutated PrP was converted into PrP as judged by Western blotting. These findings raise the possibility that the N2a cell line can support replication of different strains of prions.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-10-2555
2000-10-01
2020-09-30
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/10/0812555a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-10-2555&mimeType=html&fmt=ahah

References

  1. Bessen R. A., Marsh R. F.. 1994; Distinct PrP properties suggest the molecular basis of strain variation in transmissible mink encephalopathy. Journal of Virology68:7859–7868
    [Google Scholar]
  2. Bosque P. J., Prusiner S. B.. 2000; Cultured cell sublines highly susceptible to prion infection. Journal of Virology74:4377–4386
    [Google Scholar]
  3. Bruce M. E., Dickinson A. G.. 1987; Biological evidence that scrapie agent has an independent genome. Journal of General Virology68:79–89
    [Google Scholar]
  4. Bruce M. E., McBride P. A., Farquhar C. F.. 1989; Precise targeting of the pathology of the sialoglycoprotein, PrP, and vacuolar degeneration in mouse scrapie. Neuroscience Letters102:1–6
    [Google Scholar]
  5. Butler D. A., Scott M. R. D., Bockman J. M., Borchelt D. R., Taraboulos A., Hsiao K. K., Kingsbury D. T., Prusiner S. B.. 1988; Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins. Journal of Virology62:1558–1564
    [Google Scholar]
  6. Chandler R. L.. 1961; Encephalopathy in mice produced by inoculation with scrapie brain material. Lanceti:1378–1379
    [Google Scholar]
  7. Collinge J., Sidle K. C. L., Meads J., Ironside J., Hill A. F.. 1996; Molecular analysis of prion strain variation and the aetiology of ‘new variant’ CJD. Nature383:685–690
    [Google Scholar]
  8. DeArmond S. J., Sánchez H., Yehiely F., Qiu Y., Ninchak-Casey A., Daggett V., Camerino A. P., Cayetano J., Rogers M., Groth D., Torchia M., Tremblay P., Scott M. R., Cohen F. E., Prusiner S. B.. 1997; Selective neuronal targeting in prion disease. Neuron19:1337–1348
    [Google Scholar]
  9. Dickinson A. G., Meikle V. M.. 1969; A comparison of some biological characteristics of the mouse-passaged scrapie agents, 22A and ME7. Genetic Research13:213–225
    [Google Scholar]
  10. Endo T., Groth D., Prusiner S. B., Kobata A.. 1989; Diversity of oligosaccharide structures linked to asparagines of the scrapie prion protein. Biochemistry28:8380–8388
    [Google Scholar]
  11. Farquhar C. F., Dornan J., Moore R. C., Somerville R. A., Tunstall A. M., Hope J.. 1996; Protease-resistant PrP deposition in brain and non-central nervous system tissues of a murine model of bovine spongiform encephalopathy. Journal of General Virology77:1941–1946
    [Google Scholar]
  12. Goldfarb L. G., Petersen R. B., Tabaton M., Brown P., LeBlanc A. C., Montagna P., Cortelli P., Julien J., Vital C., Pendelbury W. W.. 1992; Fatal familial insomnia and familial Creutzfeldt–Jakob disease: disease phenotype determined by a DNA polymorphism. Science258:806–808
    [Google Scholar]
  13. Hecker R., Taraboulos A., Scott M., Pan K.-M., Torchia M., Jendroska K., DeArmond S. J., Prusiner S. B.. 1992; Replication of distinct scrapie prion isolates is region specific in brains of transgenic mice and hamsters. Genes & Development6:1213–1228
    [Google Scholar]
  14. Hill A. F., Butterworth R. J., Joiner S., Jackson G., Rossor M. N., Thomas D. J., Frosh A., Tolley N., Bell J. E., Spencer M., King A., Al-Sarraj S., Ironside J. W., Lantos P. L., Collinge J.. 1999; Investigation of variant Creutzfeldt–Jakob disease and other human prion diseases with tonsil biopsy samples. Lancet353:183–189
    [Google Scholar]
  15. Kaneko K., Zulianello L., Scott M., Cooper C. M., Wallace A. C., James T. L., Cohen F. E., Prusiner S. B.. 1997; Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation. Proceedings of the National Academy of Sciences USA94:10069–10074
    [Google Scholar]
  16. Kascsak R. J., Rubenstein R., Merz P. A., Tonna-DeMasi M., Fersko R., Carp R. I., Wisniewski H. M., Diringer H.. 1987; Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins. Journal of Virology61:3688–3693
    [Google Scholar]
  17. Kellings K., Prusiner S. B., Riesner D.. 1994; Nucleic acids in prion preparations: unspecific background or essential component?. Philosophical Transactions of the Royal Society of London B Biological Sciences343:425–430
    [Google Scholar]
  18. Lehmann S., Harris D. A.. 1997; Blockade of glycosylation promotes acquisition of scrapie-like properties by the prion protein in cultured cells. Journal of Biological Chemistry272:21479–21487
    [Google Scholar]
  19. Locht C., Chesebro B., Race R., Keith J. M.. 1986; Molecular cloning and complete sequence of prion protein cDNA from mouse brain infected with the scrapie agent. Proceedings of the National Academy of Sciences, USA83:6372–6376
    [Google Scholar]
  20. Lugaresi E., Medori R., Montagna P., Baruzzi A., Cortelli P., Lugaresi A., Tinuper P., Zucconi M., Gambetti P.. 1986; Fatal familial insomnia and dysautonomia with selective degeneration of thalamic nuclei. New England Journal of Medicine315:997–1003
    [Google Scholar]
  21. Mastrianni J. A., Nixon R., Layzer R., Telling G. C., Han D., DeArmond S. J., Prusiner S. B.. 1999; Prion protein conformation in a patient with sporadic fatal insomnia. New England Journal of Medicine340:1630–1638
    [Google Scholar]
  22. Nishida N., Harris D. A., Vilette D., Laude H., Frobert Y., Grassi J., Casanova D., Milhavet O., Lehmann S.. 2000; Successful transmission of three mouse-adapted scrapie strains to murine neuroblastoma cell lines overexpressing wild-type mouse prion protein. Journal of Virology74:320–325
    [Google Scholar]
  23. Parchi P., Capellari S., Chen S. G., Petersen R. B., Gambetti P., Kopp P., Brown P., Kitamoto T., Tateishi J., Giese A., Kretzschmar H.. 1997; Typing prion isoforms. Nature386:232–233
    [Google Scholar]
  24. Parchi P., Capellari S., Chin S., Schwarz H. B., Schecter N. P., Butts J. D., Hudkins P., Burns D. K., Powers J. M., Gambetti P.. 1999; A subtype of sporadic prion disease mimicking fatal familial insomnia. Neurology52:1757–1763
    [Google Scholar]
  25. Pattison I. H., Millson G. C.. 1961; Scrapie produced experimentally in goats with special reference to the clinical syndrome. Journal of Comparative Pathology71:101–108
    [Google Scholar]
  26. Priola S. A., Caughey B., Race R. E., Chesebro B.. 1994; Heterologous PrP molecules interfere with accumulation of protease-resistant PrP in scrapie-infected murine neuroblastoma cells. Journal of Virology68:4873–4878
    [Google Scholar]
  27. Prusiner S. B.. 1998; Prions. Proceedings of the National Academy of Sciences, USA95:13363–13383
    [Google Scholar]
  28. Race R. E., Fadness L. H., Chesebro B.. 1987; Characterization of scrapie infection in mouse neuroblastoma cells. Journal of General Virology68:1391–1399
    [Google Scholar]
  29. Rogers M., Taraboulos A., Scott M., Groth D., Prusiner S. B.. 1990; Intracellular accumulation of the cellular prion protein after mutagenesis of its Asn-linked glycosylation sites. Glycobiology1:101–109
    [Google Scholar]
  30. Rubenstein R., Merz P. A., Kascsak R. J., Scalici C. L., Papini M. C., Carp R. I., Kimberlin R. H.. 1991; Scrapie-infected spleens: analysis of infectivity, scrapie-associated fibrils, and protease-resistant proteins. Journal of Infectious Diseases164:29–35
    [Google Scholar]
  31. Rudd P. M., Endo T., Colominas C., Groth D., Wheeler S. F., Harvey D. J., Wormald M. R., Serban H., Prusiner S. B., Kobata A., Dwek R. A.. 1999; Glycosylation differences between the normal and pathogenic prion protein isoforms. Proceedings of the National Academy of Sciences, USA96:13044–13049
    [Google Scholar]
  32. Safar J., Wille H., Itri V., Groth D., Serban H., Torchia M., Cohen F. E., Prusiner S. B.. 1998; Eight prion strains have PrPSc molecules with different conformations. Nature Medicine4:1157–1165
    [Google Scholar]
  33. Schätzl H. M., Laszlo L., Holtzman D. M., Tatzelt J., DeArmond S. J., Weiner R. I., Mobley W. C., Prusiner S. B.. 1997; A hypothalamic neuronal cell line persistently infected with scrapie prions exhibits apoptosis. Journal of Virology71:8821–8831
    [Google Scholar]
  34. Scott M. R., Köhler R., Foster D., Prusiner S. B.. 1992; Chimeric prion protein expression in cultured cells and transgenic mice. Protein Science1:986–997
    [Google Scholar]
  35. Scott M., Groth D., Foster D., Torchia M., Yang S.-L., DeArmond S. J., Prusiner S. B.. 1993; Propagation of prions with artificial properties in transgenic mice expressing chimeric PrP genes. Cell73:979–988
    [Google Scholar]
  36. Scott M. R., Groth D., Tatzelt J., Torchia M., Tremblay P., DeArmond S. J., Prusiner S. B.. 1997; Propagation of prion strains through specific conformers of the prion protein. Journal of Virology71:9032–9044
    [Google Scholar]
  37. Somerville R. A.. 1999; Host and transmissible spongiform encephalopathy agent strain control glycosylation of PrP. Journal of General Virology80:1865–1872
    [Google Scholar]
  38. Taraboulos A., Rogers M., Borchelt D. R., McKinley M. P., Scott M., Serban D., Prusiner S. B.. 1990; Acquisition of protease resistance by prion proteins in scrapie-infected cells does not require asparagine-linked glycosylation. Proceedings of the National Academy of Sciences, USA87:8262–8266
    [Google Scholar]
  39. Telling G. C., Scott M., Mastrianni J., Gabizon R., Torchia M., Cohen F. E., DeArmond S. J., Prusiner S. B.. 1995; Prion propagation in mice expressing human and chimeric PrP transgenes implicates the interaction of cellular PrP with another protein. Cell83:79–90
    [Google Scholar]
  40. Telling G. C., Parchi P., DeArmond S. J., Cortelli P., Montagna P., Gabizon R., Mastrianni J., Lugaresi E., Gambetti P., Prusiner S. B.. 1996; Evidence for the conformation of the pathologic isoform of the prion protein enciphering and propagating prion diversity. Science274:2079–2082
    [Google Scholar]
  41. Zulianello L., Kaneko K., Scott M., Erpel S., Han D., Cohen F. E., Prusiner S. B.. 2000; Dominant negative inhibition of prion formation diminished by deletion mutagenesis of the prion protein. Journal of Virology74:4351–4360
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-10-2555
Loading
/content/journal/jgv/10.1099/0022-1317-81-10-2555
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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