Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

RNA molecules stimulate prion protein conversion

Abstract

Much evidence supports the hypothesis that the infectious agents of prion diseases are devoid of nucleic acid, and instead are composed of a specific infectious protein1. This protein, PrPSc, seems to be generated by template-induced conformational change of a normally expressed glycoprotein, PrPC (ref. 2). Although numerous studies have established the conversion of PrPC to PrPSc as the central pathogenic event of prion disease, it is unknown whether cellular factors other than PrPC might be required to stimulate efficient PrPSc production. We investigated the biochemical amplification of protease-resistant PrPSc-like protein (PrPres) using a modified version3 of the protein-misfolding cyclic amplification method4. Here we report that stoichiometric transformation of PrPC to PrPres in vitro requires specific RNA molecules. Notably, whereas mammalian RNA preparations stimulate in vitro amplification of PrPres, RNA preparations from invertebrate species do not. Our findings suggest that host-encoded stimulatory RNA molecules may have a role in the pathogenesis of prion disease. They also provide a practical approach to improve the sensitivity of diagnostic techniques based on PrPres amplification.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Effect of various enzymes on PrPres amplification.
Figure 2: Nucleases do not cause proteolytic, steric or end-product inhibition of PrPres amplification.
Figure 3: Reconstitution of PrPres amplification with RNA.
Figure 4: Stimulation of PrPres amplification with RNA.

References

  1. 1

    Prusiner, S. B. Novel proteinaceous infectious particles cause scrapie. Science 216, 136–144 (1982)

    CAS  PubMed  PubMed Central  Google Scholar 

  2. 2

    Prusiner, S. B. (ed.) Prion Biology and Diseases (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1999)

  3. 3

    Lucassen, R., Nishina, K. & Supattapone, S. In vitro amplification of protease-resistant prion protein requires free sulfhydryl groups. Biochemistry 42, 4127–4135 (2003)

    CAS  Article  Google Scholar 

  4. 4

    Saborio, G. P., Permanne, B. & Soto, C. Sensitive detection of pathological prion protein by cyclic amplification of protein misfolding. Nature 411, 810–813 (2001)

    CAS  Article  Google Scholar 

  5. 5

    Lockard, R. E. & Kumar, A. Mapping tRNA structure in solution using double-strand-specific ribonuclease V1 from cobra venom. Nucleic Acids Res. 9, 5125–5140 (1981)

    CAS  Article  Google Scholar 

  6. 6

    Banks, G. R. A ribonuclease H from Ustilago maydis. Properties, mode of action and substrate specificity of the enzyme. Eur. J. Biochem. 47, 499–507 (1974)

    CAS  Article  Google Scholar 

  7. 7

    Kocisko, D. A. et al. Cell-free formation of protease-resistant prion protein. Nature 370, 471–474 (1994)

    CAS  Article  Google Scholar 

  8. 8

    Caughey, B., Horiuchi, M., Demaimay, R. & Raymond, G. J. Assays of protease-resistant prion protein and its formation. Methods Enzymol. 309, 122–133 (1999)

    CAS  Article  Google Scholar 

  9. 9

    Derrington, E. et al. PrPC has nucleic acid chaperoning properties similar to the nucleocapsid protein of HIV-1. C. R. Acad. Sci. III 325, 17–23 (2002)

    CAS  Google Scholar 

  10. 10

    Moscardini, M. et al. Functional interactions of nucleocapsid protein of feline immunodeficiency virus and cellular prion protein with the viral RNA. J. Mol. Biol. 318, 149–159 (2002)

    CAS  Article  Google Scholar 

  11. 11

    Gabus, C. et al. The prion protein has RNA binding and chaperoning properties characteristic of nucleocapsid protein NCP7 of HIV-1. J. Biol. Chem. 276, 19301–19309 (2001)

    CAS  Article  Google Scholar 

  12. 12

    Gabus, C. et al. The prion protein has DNA strand transfer properties similar to retroviral nucleocapsid protein. J. Mol. Biol. 307, 1011–1021 (2001)

    CAS  Article  Google Scholar 

  13. 13

    Nandi, P. K., Leclerc, E., Nicole, J. C. & Takahashi, M. DNA-induced partial unfolding of prion protein leads to its polymerisation to amyloid. J. Mol. Biol. 322, 153–161 (2002)

    CAS  Article  Google Scholar 

  14. 14

    Cordeiro, Y. et al. DNA converts cellular prion protein into the β-sheet conformation and inhibits prion peptide aggregation. J. Biol. Chem. 276, 49400–49409 (2001)

    CAS  Article  Google Scholar 

  15. 15

    Weissmann, C. A ‘unified theory’ of prion propagation. Nature 352, 679–683 (1991)

    CAS  Article  Google Scholar 

  16. 16

    Chapon, C., Cech, T. R. & Zaug, A. J. Polyadenylation of telomerase RNA in budding yeast. RNA 3, 1337–1351 (1997)

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors thank G. Saborio, C. Soto, V. Ambros, C. Cole and W. Wickner for helpful advice. This work was supported by the Burroughs Wellcome Fund Career Development Award, the Hitchcock Foundation, and an NIH Clinical Investigator Development Award.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Surachai Supattapone.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Deleault, N., Lucassen, R. & Supattapone, S. RNA molecules stimulate prion protein conversion. Nature 425, 717–720 (2003). https://doi.org/10.1038/nature01979

Download citation

Further reading

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing