Analysis of nucleic acids in purified scrapie prion preparations

  • K. Kellings
  • N. Meyer
  • C. Mirenda
  • S. B. Prusiner
  • D. Riesner
Part of the Archives of Virology book series (ARCHIVES SUPPL, volume 7)


Amount, type, and size of nucleic acid molecules associated with purified prion preparations were analyzed. Return refocusing gel electrophoresis (RRGE) was developed to detect homogeneous and heterogeneous nucleic acids extracted from highly purified scrapie prion preparations. With this method all types of nucleic acids in the size range from 13 to several thousand nucleotides could be analyzed. The recovery of all nucleic acids, after deproteinization and two-phase extraction was higher than 90%. Despite extensive nuclease digestions some small polynucleotides remained. Although a scrapie-specific nucleic acid cannot be excluded, the results further define the possible characteristics for such a hypothetical molecule. If it was homogeneous in size, then it would be <80 nt in length at a particle-to-infectivity ratio (P/I) near unity; if the other extreme, i.e. totally heterogeneous scrapiespecific nucleic acids were assumed, then scrapie-specific nucleic acids would have to include molecules smaller than 240 nt. In order to exclude the possibility that unspecific background nucleic acid is entrapped in prion-rods, infectious material has to be prepared without a proteolysis and rod formation, and the analysis of nucleic acids performed with those preparations.


Nucleic Acid Prion Protein Micrococcal Nuclease Prion Infectivity Potato Spindle Tuber Viroid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Alper T, Haig DA, Clarke MC (1966) The exceptionally small size of the scrapie agent. Biochem Biophys Res Commun 22: 278–284PubMedCrossRefGoogle Scholar
  2. 2.
    Alper T, Cramp WA, Haig DA, Clarke MC (1967) Does the agent of scrapie replicate without nucleic acids? Nature 214: 764–766PubMedCrossRefGoogle Scholar
  3. 3.
    Prusiner SB (1982) Novel proteinaceous infectious particles cause scrapie. Science 216: 136–144PubMedCrossRefGoogle Scholar
  4. 4.
    Riesner D (1991) The search for a nucleic acid component to scapie infectivity. Semin Virol 2: 215–226Google Scholar
  5. 5.
    Bruce ME, Dickinson AG (1987) Biological evidence that scrapie agent has an independent genome. J Gen Virol 68: 79–89PubMedCrossRefGoogle Scholar
  6. 6.
    Kimberlin RH, Cole S, Walker DA (1987) Temporary and permanent modifications to a single strain of mouse scrapie on transmission to rats and hamsters. J Gen Virol 68: 1875–1881PubMedCrossRefGoogle Scholar
  7. 7.
    Prusiner SB (1991) Molecular biology of prion diseases. Science 252: 1515–1522PubMedCrossRefGoogle Scholar
  8. 8.
    Weissmann C (1991) A “unified theory” of prion propagation. Nature 352: 679–683PubMedCrossRefGoogle Scholar
  9. 9.
    Meyer N, Rosenbaum V, Schmidt B, Gilles K, Mirenda C, Groth D, Prusiner SB, Riesner D (1991) Search for a putative scrapie genome in purified prion fractions reveals a paucity of nucleic acids. J Gen Virol 72: 37–49PubMedCrossRefGoogle Scholar
  10. 10.
    Kellings K, Meyer N, Mirenda C, Prusiner SB, Riesner D (1992) Further analysis of nucleic acids in purified scrapie prion preparations by an improved return refocusing gel electrophoresis ( RRGE ). J Gen Virol 73: 1025–1029PubMedCrossRefGoogle Scholar
  11. 11.
    Gabizon R, McKinley MP, Prusiner SB (1987) Purified prion proteins and scrapie infectivity copartition into liposomes. Proc Natl Acad Sci USA 84: 4017–4021PubMedCrossRefGoogle Scholar
  12. 12.
    Gabizon R, Groth DF, McKinley MP, Prusiner SB (1988) Immunoaffinity purification and neutralization of scrapie prion infectivity. Proc Natl Acad Sci USA 85: 6617–6621PubMedCrossRefGoogle Scholar
  13. 13.
    Prusiner SB, Cochran SP, Groth DF, Downey DE, Bowman KA, Martinez HM (1982) Measurement of the scrapie agent using an incubation time interval assay. Ann Neurol 11: 353–358PubMedCrossRefGoogle Scholar
  14. 14.
    Haseloff J, Mohamed NA, Symons RH (1982) Viroid RNAs of cadang-cadang disease of coconuts. Nature 299: 316–322CrossRefGoogle Scholar
  15. 15.
    Palukaitis P, Zaitlin M (1987) The nature and biological significance of linear potato spindle tuber viroid molecules. Virology 157: 199–210PubMedCrossRefGoogle Scholar
  16. 16.
    Tabler M, Sänger HL (1984) Cloned single-and double-stranded DNA copies of potato spindle tuber viroid ( PSTV) RNA and co-inoculated subgenomic DNA fragments are infectious. EMBO J 3: 3055–2062PubMedGoogle Scholar
  17. 17.
    Scott M, Foster D, Mirenda C, Serban D, Coufal F, Wälchli M, Torchia M, Groth D, Carlson G, DeArmond SJ, Westaway D, Prusiner, SB (1989) Transgenic mice expressing hamster prion protein produce species-specific scrapie infectivity and amyloid plaques. Cell 59: 847–857PubMedCrossRefGoogle Scholar
  18. 18.
    Hsiao KK, Scott M, Foster D, Groth D, DeArmond SJ, Prusiner SB (1990) Spontaneous neurodegeneration in transgenic mice with mutant prion protein. Science 250: 1587–1590PubMedCrossRefGoogle Scholar
  19. 19.
    McKinley MP, Meyer R, Kenaga L, Rahbar F, Cotter R, Serban A, Prusiner SB (1991) Scrapie prion rod formation in vitro requires both detergent extraction and limited proteolysis. J Virol 65: 1440–1449Google Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • K. Kellings
    • 1
  • N. Meyer
    • 1
  • C. Mirenda
    • 2
  • S. B. Prusiner
    • 2
  • D. Riesner
    • 1
    • 3
  1. 1.Institut für Physikalische BiologieHeinrich-Heine-Universität DüsseldorfDüsseldorfFederal Republic of Germany
  2. 2.Department of NeurologyUniversity of CaliforniaSan FranciscoUSA
  3. 3.Institut für Physikalische BiologieHeinrich-Heine-Universität DüsseldorfDüsseldorfFederal Republic of Germany

Personalised recommendations