1887

Journal of General Virology header logo

Volume 83, Issue 5

Mutant prion protein acquires resistance to protease in mouse neuroblastoma cells Free

Abstract

Conversion of the cellular isoform of the prion protein (PrP) into the pathogenic isoform (PrP) is thought to be the causative event in prion diseases. Biochemically, PrP differs from PrP in its partial resistance to proteinase K (PK). The amino acid sequence AGAAAAGA, comprising residues 112–119 of the murine PrP, has been shown to be amyloidogenic and evolutionarily conserved. To assess the effect of mutations at and around this hydrophobic sequence on protease resistance, the sequence was replaced either by alanines or by glycines and, in a third mutant, a large part surrounding this region was removed. The PrP mutant carrying substitutions of glycines for alanines showed PK resistance and aberrant proteolytic processing. Tetracycline-induced expression of this mutant indicated that resistance to protease is acquired concurrent with the synthesis of the protein. These findings indicate that mutations in the central hydrophobic region lead to immediate alterations in PrP structure and processing.

  • Received:
  • Accepted:
  • Published Online:
© Microbiology Society
Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-83-5-1237
2002-05-01
2024-04-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/83/5/0831237a.html?itemId=/content/journal/jgv/10.1099/0022-1317-83-5-1237&mimeType=html&fmt=ahah

References

  1. Borchelt D. R., Scott D., Taraboulos A., Stahl N., Prusiner S. B. 1990; Scrapie and cellular prion proteins differ in their kinetics of synthesis and topology in cultured cells. Journal of Cell Biology 110:743–752
     [Google Scholar]
  2. Brandner S., Isenmann S., Raeber A., Fischer M., Sailer A., Kobayashi Y., Marino S., Weissmann C., Aguzzi A. 1996; Normal host prion protein necessary for scrapie-induced neurotoxicity. Nature 379:339–343
     [Google Scholar]
  3. Brown D. R. 2000a; Prion protein peptides: optimal toxicity and peptide blockade of toxicity. Molecular and Cellular Neurosciences 15:66–78
     [Google Scholar]
  4. Brown D. R. 2000b; PrPSc-like prion protein peptide inhibits the function of cellular prion protein. Biochemical Journal 352:511–518
     [Google Scholar]
  5. Brown D. R., Schmidt B., Kretzschmar H. A. 1996; Role of microglia and host prion protein in neurotoxicity of prion protein fragment. Nature 380:345–347
     [Google Scholar]
  6. Brown D. R., Qin K., Herms J. W., Madlung A., Manson J., Strome R., Fraser P. E., Kruck T., von Bohlen A., Schulz-Schaeffer W., Giese A., Westaway D., Kretzschmar H. 1997a; The cellular prion protein binds copper in vivo . Nature 390:684–687
     [Google Scholar]
  7. Brown D. R., Schulz-Schaeffer W. J., Schmidt B., Kretzschmar H. A. 1997b; Prion protein-deficient cells show altered response to oxidative stress due to decreased SOD-1 activity. Experimental Neurology 146:104–112
     [Google Scholar]
  8. Büeler H., Aguzzi A., Sailer A., Greiner R. A., Autenried P., Aguet M., Weissmann C. 1993; Mice devoid of PrP are resistant to scrapie. Cell 73:1339–1347
     [Google Scholar]
  9. Büeler H., Raeber A., Sailer A., Fischer M., Aguzzi A., Weissmann C. 1994; High prion and PrPSc levels but delayed onset of disease in scrapie-inoculated mice heterozygous for a disrupted PrP gene. Molecular Medicine 1:19–30
     [Google Scholar]
  10. Caughey B., Race R. E., Ernst D., Buchmeier M. J., Chesebro B. 1989; Prion protein biosynthesis in scrapie-infected and uninfected neuroblastoma cells. Journal of Virology 63:175–181
     [Google Scholar]
  11. Caughey B. W., Dong A., Bhat K. S., Ernst D., Hayes S. F., Caughey W. S. 1991; Secondary structure analysis of the scrapie-associated protein PrP27–30 in water by infrared spectroscopy. Biochemistry 30:7672–7680
     [Google Scholar]
  12. Chabry J., Caughey B., Chesebro B. 1998; Specific inhibition of in vitro formation of protease-resistant prion protein by synthetic peptides. Journal of Biological Chemistry 273:13203–13207
     [Google Scholar]
  13. Chen S. G., Teplow D. B., Parchi P., Teller J. K., Gambetti P., Autilio-Gambetti L. 1995; Truncated forms of the human prion protein in normal brain and in prion diseases. Journal of Biological Chemistry 270:19173–19180
     [Google Scholar]
  14. Daude N., Lehmann S., Harris D. A. 1997; Identification of intermediate steps in the conversion of a mutant prion protein to a scrapie-like form in cultured cells. Journal of Biological Chemistry 272:11604–11612
     [Google Scholar]
  15. Donne D. G., Viles J. H., Groth D., Mehlhorn I., James T. L., Cohen F. E., Prusiner S. B., Wright P. E., Dyson H. J. 1997; Structure of recombinant full-length hamster prion protein PrP(29–231): the N terminus is highly flexible. Proceedings of the National Academy of Sciences, USA 94:13452–13457
     [Google Scholar]
  16. Forloni G., Angeretti N., Chiesa R., Monzani E., Salmona M., Bugiani O., Tagliavini F. 1993; Neurotoxicity of a prion protein fragment. Nature 362:543–546
     [Google Scholar]
  17. Gasset M., Baldwin M. A., Lloyd D. H., Gabriel J.-M., Holtzman D. M., Cohen F., Fletterick R., Prusiner S. B. 1992; Predicted α-helical regions of the prion protein when synthesized as peptides form amyloid. Proceedings of the National Academy of Sciences, USA 89:10940–10944
     [Google Scholar]
  18. Haraguchi T., Fisher S., Olofsson S., Endo T., Groth D., Tarentino A., Borchelt D. R., Teplow D., Hood L., Burlingame A., Lycke E., Kobata A., Prusiner S. B. 1989; Asparagine-linked glycosylation of the scrapie and cellular prion proteins. Archives of Biochemistry and Biophysics 274:1–13
     [Google Scholar]
  19. Harris D. A., Huber M. T., van Dijken P., Shyng S. L., Chait B. T., Wang R. 1993; Processing of a cellular prion protein: identification of N- and C-terminal cleavage sites. Biochemistry 32:1009–1016
     [Google Scholar]
  20. Hegde R. S., Mastrianni J. A., Scott M. R., DeFea K. A., Tremblay P., Torchia M., DeArmond S. J., Prusiner S. B., Lingappa V. R. 1998; A transmembrane form of the prion protein in neurodegenerative disease. Science 279:827–834
     [Google Scholar]
  21. Hegde R. S., Tremblay P., Groth D., DeArmond S. J., Prusiner S. B., Lingappa V. R. 1999; Transmissible and genetic prion diseases share a common pathway of neurodegeneration. Nature 402:822–826
     [Google Scholar]
  22. Herms J., Tings T., Gall S., Madlung A., Giese A., Siebert H., Schürmann P., Windl O., Brose N., Kretzschmar H. 1999; Evidence of presynaptic location and function of the prion protein. Journal of Neuroscience 19:8866–8875
     [Google Scholar]
  23. Higuchi R., Krummel B., Saiki R. K. 1988; A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Research 16:7351–7367
     [Google Scholar]
  24. Hölscher C., Delius H., Bürkle A. 1998; Overexpression of nonconvertible PrPcΔ114–121 in scrapie-infected mouse neuroblastoma cells leads to trans -dominant inhibition of wild-type PrPSc accumulation. Journal of Virology 72:1153–1159
     [Google Scholar]
  25. Hornemann S., Korth C., Oesch B., Riek R., Wider G., Wüthrich K., Glockshuber R. 1997; Recombinant full-length murine prion protein, mPrP(23–231): purification and spectroscopic characterization. FEBS Letters 413:277–281
     [Google Scholar]
  26. Ivanova L., Barmada S., Kummer T., Harris D. A. 2001; Mutant prion proteins are partially retained in the endoplasmic reticulum. Journal of Biological Chemistry 276:42409–42421
     [Google Scholar]
  27. Jiménez-Huete A., Lievens P. M. J., Vidal R., Piccardo P., Ghetti B., Tagliavini F., Frangione B., Prelli F. 1998; Endogenous proteolytic cleavage of normal and disease-associated isoforms of the human prion protein in neural and non-neural tissues. American Journal of Pathology 153:1561–1572
     [Google Scholar]
  28. Klebe R. J., Ruddle F. H. 1969; Neuroblastoma: cell culture analysis of a differentiating stem cell system. Journal of Cell Biology 43: 69a
    [Google Scholar]
  29. Korth C., Stierli B., Streit P., Moser M., Schaller O., Fischer R., Schulz-Schaeffer W., Kretzschmar H., Raeber A., Braun U., Ehrensperger F., Hornemann S., Glockshuber R., Riek R., Billeter M., Wüthrich K., Oesch B. 1997; Prion (PrPSc)-specific epitope defined by a monoclonal antibody. Nature 390:74–77
     [Google Scholar]
  30. Krasemann S., Groschup M. H., Harmeyer S., Hunsmann G., Bodemer W. 1996; Generation of monoclonal antibodies against human prion proteins in PrP0/0 mice. Molecular Medicine 2:725–734
     [Google Scholar]
  31. LeGendre N. 1990; Immobilon-P transfer membrane: applications and utility in protein biochemical analysis. Biotechniques 9:788–805
     [Google Scholar]
  32. Lehmann S., Harris D. A. 1996; Mutant and infectious prion proteins display common biochemical properties in cultured cells. Journal of Biological Chemistry 271:1633–1637
     [Google Scholar]
  33. 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 Chemistry 272:21479–21487
     [Google Scholar]
  34. Liu H., Farr-Jones S., Ulyanov N. B., Llinas M., Marqusee S., Groth D., Cohen F. E., Prusiner S. B., James T. L. 1999; Solution structure of Syrian hamster prion protein rPrP(90–231). Biochemistry 38:5362–5377
     [Google Scholar]
  35. Mouillet-Richard S., Ermonval M., Chebassier C., Laplanche J. L., Lehmann S., Launay J. M., Kellermann O. 2000; Signal transduction through prion protein. Science 289:1925–1928
     [Google Scholar]
  36. Muramoto T., Scott M., Cohen F. E., Prusiner S. B. 1996; Recombinant scrapie-like prion protein of 106 amino acids is soluble. Proceedings of the National Academy of Sciences, USA 93:15457–15462
     [Google Scholar]
  37. Pan K. M., Stahl N., Prusiner S. B. 1992; Purification and properties of the cellular prion protein from Syrian hamster brain. Protein Science 1:1343–1352
     [Google Scholar]
  38. Pan K.-M., Baldwin M., Nguyen J., Gasset M., Serban A., Groth D., Mehlhorn I., Huang Z., Fletterick R. J., Cohen F. E., Prusiner S. B. 1993; Conversion of α-helices into β-sheets features in the formation of the scrapie prion proteins. Proceedings of the National Academy of Sciences, USA 90:10962–10966
     [Google Scholar]
  39. Petersen R. B., Parchi P., Richardson S. L., Urig C. B., Gambetti P. 1996; Effect of the D178N mutation and the codon 129 polymorphism on the metabolism of the prion protein. Journal of Biological Chemistry 271:12661–12668
     [Google Scholar]
  40. Priola S. A., Chesebro B. 1998; Abnormal properties of prion protein with insertional mutations in different cell types. Journal of Biological Chemistry 273:11980–11985
     [Google Scholar]
  41. Prusiner S. B., DeArmond S. J. 1994; Prion diseases and neurodegeneration. Annual Review of Neuroscience 17:311–339
     [Google Scholar]
  42. Prusiner S. B., Telling G., Cohen F. E., DeArmond S. J. 1996; Prion diseases of humans and animals. Seminars in Virology 7:159–173
     [Google Scholar]
  43. Quaglio E., Chiesa R., Harris D. A. 2001; Copper converts the cellular prion protein into a protease-resistant species that is distinct from the scrapie isoform. Journal of Biological Chemistry 276:11432–11438
     [Google Scholar]
  44. Riek R., Hornemann S., Wider G., Billeter M., Glockshuber R., Wüthrich K. 1996; NMR structure of the mouse prion protein domain PrP(121–231). Nature 382:180–182
     [Google Scholar]
  45. Riek R., Hornemann S., Wider G., Glockshuber R., Wüthrich K. 1997; NMR characterization of the full-length recombinant murine prion protein, mPrP(23–231). FEBS Letters 413:282–288
     [Google Scholar]
  46. Rogers M., Serban D., Gyuris T., Scott M., Torchia T., Prusiner S. B. 1991; Epitope mapping of the Syrian hamster prion protein utilizing chimeric and mutant genes in a vaccinia virus expression system. Journal of Immunology 147:3568–3574
     [Google Scholar]
  47. Safar J., Roller P. P., Gadjusek D. C., Gibbs C. J.Jr(1993; Conformational transitions, dissociation, and unfolding of scrapie amyloid (prion) protein. Journal of Biological Chemistry 268:20276–20284
     [Google Scholar]
  48. Scott M., Foster D., Mirenda C., Serban D., Coufal F., Wälchli M., Torchia M., Groth D., Carlson G., DeArmond S. J., Westaway D., Prusiner S. B. 1989; Transgenic mice expressing hamster prion protein produce species-specific scrapie infectivity and amyloid plaques. Cell 59:847–857
     [Google Scholar]
  49. Shyng S. L., Huber M. T., Harris D. A. 1993; A prion protein cycles between the cell surface and an endocytic compartment in cultured neuroblastoma cells. Journal of Biological Chemistry 268:15922–15928
     [Google Scholar]
  50. Singh N., Zanusso G., Chen S. G., Fujioka H., Richardson S., Gambetti P., Petersen R. B. 1997; Prion protein aggregation reverted by low temperature in transfected cells carrying a prion protein gene mutation. Journal of Biological Chemistry 272:28461–28470
     [Google Scholar]
  51. Stewart R. S., Harris D. A. 2001; Most pathogenic mutations do not alter the membrane topology of the prion protein. Journal of Biological Chemistry 276:2212–2220
     [Google Scholar]
  52. Tremblay P., Meiner Z., Galou M., Heinrich C., Petromilli C., Lisse T., Cayetano J., Torchia M., Mobley W., Bujard H., DeArmond S. J., Prusiner S. B. 1998; Doxycycline control of prion protein transgene expression modulates prion disease in mice. Proceedings of the National Academy of Sciences, USA 95:12580–12585
     [Google Scholar]
  53. Viles J. H., Cohen F. E., Prusiner S. B., Goodin D. B., Wright P. E., Dyson H. J. 1999; Copper binding to the prion protein: structural implications of four identical cooperative binding sites. Proceedings of the National Academy of Sciences, USA 96:2042–2047
     [Google Scholar]
  54. Windl O., Lorenz H., Behrens C., Römer A., Kretzschmar H. A. 1999; Construction and characterization of murine neuroblastoma cell clones allowing inducible and high expression of the prion protein. Journal of General Virology 80:15–21
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-83-5-1237
Loading
Mutant prion protein acquires resistance to protease in mouse neuroblastoma cells
J. Gen. Virol. 83, 1237 (2002); https://doi.org/10.1099/0022-1317-83-5-1237
/content/journal/jgv/10.1099/0022-1317-83-5-1237
/content/journal/jgv/10.1099/0022-1317-83-5-1237
Loading

Data & Media loading...

Most cited 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