1887

Abstract

The purpose of this study was to investigate previous reports of a scrapie-specific 1.2 kb single-stranded DNA observed in alkaline agarose electrophoresis gels. Protocols were developed to be as consistent as possible with those used previously. Partial subcellular fractionation was applied to the brains of hamsters clinically affected by the 263K strain of scrapie. Nucleic acids were then isolated, and compared electrophoretically to nucleic acids isolated from equivalent fractions, made from the brains of hamsters inoculated with normal brain. Several modifications to the protocols were suggested. Results obtained by using these modifications were compared to results obtained using the original protocols. Scrapie-specific DNA was not observed in a total of eleven consecutive experiments. Thus, these results do not support the hypothesis that the agents of transmissible spongiform encephalopathy contain a single-stranded 1.2 kb DNA species as previously described.

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1996-09-01
2024-04-20
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References

  1. Adams D. H. 1972; Studies on DNA from normal and scrapie affected mouse brain. Journal of Neurochemistry 19:1869–1882
    [Google Scholar]
  2. Aitken J. M., Williamson J. L., Marsh R. F. 1989; Evidence of mitochondrial involvement in scrapie infection. Journal of Virology 63:1686–1694
    [Google Scholar]
  3. Aiken J. M., Williamson J. L., Borchardt L. M., Marsh R. F. 1990; Presence of mitochondrial D-loop DNA in scrapie infected brain preparations enriched for the prion protein. Journal of Virology 64:3265–3268
    [Google Scholar]
  4. Akowitz A., Sklaviadis T., Manuelidis L. 1994; Endogenous viral complexes with long RNA cosediment with the agent of Creutzfeldt-Jakob disease. Nucleic Acids Research 22:1101–1107
    [Google Scholar]
  5. Alper T., Cramp W. A., Haig D. A., Clarke M. C. 1967; Does the agent of scrapie replicate without nucleic acid?. Nature 214:764–766
    [Google Scholar]
  6. Basler K., Oesch B., Scott M., Westaway D., Walchli M., Groth D. F., McKinley M. P., Prusiner S. B., Weissman C. 1986; Scrapie and cellular PrP isoforms are encoded by the same chromosomal gene. Cell 46:417–428
    [Google Scholar]
  7. Bessen R. A., Kocisko D. A., Raymond G. J., Nandan S., Lansbury P. T., Caughey B. 1995; Non-genetic propagation of scrapie-specific prion protein. Nature 375:698–700
    [Google Scholar]
  8. Borras M. T., Kingsbury D. T., Gadjusek D. C., Gibbs C. J. Jr 1982; Inability to transmit scrapie by transfection of mouse embryo cells in vitro . Journal of General Virology 58:263–271
    [Google Scholar]
  9. Bortner C. D., Oldenburg N. B. E., Cidlowski J. A. 1995; The role of DNA fragmentation in apoptosis. Trends in Cell Biology 5:21–26
    [Google Scholar]
  10. Bountiff L. 1983 Nucleic acids in scrapie PhD thesis Reading University, Reading, UK;
    [Google Scholar]
  11. Bruce M., Chree A., McConnell I., Foster J., Pearson G., Fraser H. 1994; Transmission of bovine spongiform encephalopathy and scrapie to mice: strain variation and the species barrier. Philosophical Transactions of the Royal Society of London Series B Biological Sciences 343:405–411
    [Google Scholar]
  12. Duguid J. R., Dinauer M. C. 1989; Library subtraction of in vitro cDNA libraries to identify differentially expressed genes in scrapie infection. Nucleic Acids Research 18:2789–2792
    [Google Scholar]
  13. Fraser H., Dickinson A. G. 1968; The sequential development of the brain lesions in three strains of mice. Journal of Comparative Pathology 78:301–311
    [Google Scholar]
  14. Hallick R. B., Chelm B. K., Gray P. W., Orozco E. M. Jr 1977; Use of aurintricarboxylic acid as an inhibitor of nucleases during nucleic acid isolation. Nucleic Acids Research 4:3055–3064
    [Google Scholar]
  15. Hunter G. D., Kimberlin R. H., Collis S. C., Millson G. C. 1973; Viral and non-viral properties of the scrapie agent. Annals of Clinical Research 5:262–267
    [Google Scholar]
  16. Kimberlin R. H., Millson G. C., Bountiff L., Collis S. C. 1974a; A comparison of the biochemical changes induced in mouse brain by cuprizone toxicity and by scrapie infection. Journal of Comparative Pathology 84:263–270
    [Google Scholar]
  17. Kimberlin R. H., Shirt D. B., Collis S. C. 1974b; The turnover of isotopically labelled DNA in vivo in developing, adult and scrapie affected mouse brain. Journal of Neurochemistry 23:241–248
    [Google Scholar]
  18. Kimberlin R. H., Walker C. A. 1977; Characteristics of a short incubation model of scrapie in the golden hamster. Journal of General Virology 34:295–304
    [Google Scholar]
  19. Kimberlin R. H., Walker C. A. 1979; Pathogenesis of mouse scrapie: dynamics of agent replication in spleen, spinal cord and brain after infection by different routes. Journal of Comparative Pathology 89:551–562
    [Google Scholar]
  20. Lax A. J., Bountiff L., Millson G. C., Hunter G. D. 1982; Some new insights into the nature of the scrapie agent. In Virus Non-Conventionels et Affections Du Systeme Nerveaux Central pp 47–56 Edited by Court L. A., Cathala F. Paris: Masson;
    [Google Scholar]
  21. McBride P. A., Bruce M. E., Fraser H. 1988; Immunostaining of scrapie cerebral amyloid plaques with antisera raised to scrapie-associated fibrils (SAF). Neuropathology and Applied Neurohiology 14:325–336
    [Google Scholar]
  22. McKinley M. P., Bolton D. C., Prusiner S. B. 1983; A protease resistant protein is a structural component of the scrapie agent. Cell 35:57–62
    [Google Scholar]
  23. Manuelidis L., Manuelidis E. E. 1981; Search for specific DNAs in Creutzfeldt-Jakob infectious brain fractions using nick translation. Virology 109:435–443
    [Google Scholar]
  24. Meyer N., Rosenbaum V., Schmidt B., Gilles K., Mirenda C., Groth D. F., Prusiner S. B., Reisner D. 1991; Search for a putative scrapie genome in purified prion fractions reveals a paucity of nucleic acids. Journal of General Virology 72:1–113
    [Google Scholar]
  25. Millson G. C. 1965; Lysosomal enzymes in normal and scrapie mouse brain. Journal of Neurochemistry 12:461–468
    [Google Scholar]
  26. Narang H. K. 1990; Detection of single stranded DNA in scrapie infected brain by electron microscopy. journal of Molecular Biology 216:469–473
    [Google Scholar]
  27. Narang H. K. 1991; Evidence of single stranded DNA in tubulo-filamentous particles: their relationship to scrapie-associated fibrils. Intervirology 32:185–192
    [Google Scholar]
  28. Narang H. K. 1993; Molecular cloning of single stranded DNA purified from scrapie infected hamster brain. Research in Virology 144:375–387
    [Google Scholar]
  29. Narang H. K., Millar N. S., Asher D. M., Gadjusek D. C. 1991; Increased multimeric mitochondrial DNA in the brain of scrapie infected hamsters. Intervirology 32:316–324
    [Google Scholar]
  30. Norberg A. B., Persson H. 1984; Accumulation of heavy-metal ions by Zoogloea ramigera . Biotechnology and Bioengineering 26:239–246
    [Google Scholar]
  31. Prusiner S. B. 1995; Biochemistry and genetics of prion proteins. European journal of Medicinal Chemistry 30: Suppl (Is–688s)11s–50s
    [Google Scholar]
  32. Rubenstein R., Merz P. A., Kascak 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 Diseases 164:29–35
    [Google Scholar]
  33. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: A Laboratory Manual 2nd edn New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  34. Stahl N., Baldwin M. A., Teplow D. B., Hood L., Gibson B. W., Burlinghame A. L., Prusiner S. B. 1993; Structural analysis of the scrapie prion protein using mass spectrometry and amino acid sequencing. Biochemistry 32:1991–2002
    [Google Scholar]
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