Abstract
Key questions regarding the molecular nature of prions are how different prion strains can be propagated by the same protein and whether they are only protein1,2,3. Here we demonstrate the protein-only nature of prion strains in a yeast model, the [PSI] genetic element that enhances the read-through of nonsense mutations in the yeast Saccharomyces cerevisiae4,5. Infectious fibrous aggregates containing a Sup35 prion-determining amino-terminal fragment labelled with green fluorescent protein were purified from yeast harbouring distinctive prion strains. Using the infectious aggregates as ‘seeds’, elongated fibres were generated in vitro from the bacterially expressed labelled prion protein. De novo generation of strain-specific [PSI] infectivity was demonstrated by introducing sheared fibres into uninfected yeast hosts. The cross-sectional morphology of the elongated fibres generated in vitro was indistinguishable from that of the short yeast seeds, as visualized by electron microscopy. Electron diffraction of the long fibres showed the 4.7 Å spacing characteristic of the cross-beta structure of amyloids. The fact that the amyloid fibres nucleated in vitro propagate the strain-specific infectivity of the yeast seeds implies that the heritable information of distinct prion strains must be encoded by different, self-propagating cross-beta folding patterns of the same prion protein.
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Acknowledgements
We thank D. L. D. Caspar for support, suggestions and assistance with the manuscript, and C. Long and B. Patel for reviewing the manuscript. This work was supported by an NIH grant to D. L. D. Caspar and an NIH Research Service Award to C.-Y.K.
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Supplementary information
Supplementary Figure 1
His5-Sup35(1-61)-GFP-Strep(II) samples analysed by SDS-polyacrylamide gel electrophoresis. (PDF 177 kb)
Supplementary Figure 2
The morphology of protein aggregates spontaneously formed in His5-Sup35(1-61)-GFP-Strep(II) solutions. (PDF 212 kb)
Supplementary Table 1
Strain-specific transformation by [PSI] particles. (PDF 117 kb)
Supplementary Table 2
Competition among [PSI] strains. (PDF 110 kb)
Supplementary Table 3
Proteinaceous nature of [PSI]. (PDF 113 kb)
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King, CY., Diaz-Avalos, R. Protein-only transmission of three yeast prion strains. Nature 428, 319–323 (2004). https://doi.org/10.1038/nature02391
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DOI: https://doi.org/10.1038/nature02391
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