Abstract
Aggregation-prone proteins often misfold into multiple distinct amyloid conformations that dictate different physiological impacts. Although amyloid formation is triggered by a transient nucleus, the mechanism by which an initial nucleus is formed and allows the protein to form a specific amyloid conformation has been unclear. Here we show that, before fiber formation, the prion domain (Sup35NM, consisting of residues 1–254) of yeast prion Sup35, the [PSI+] protein determinant, forms oligomers in a temperature-dependent, reversible manner. Mutational and biophysical analyses revealed that 'non-native' aromatic interactions outside the amyloid core drive oligomer formation by bringing together different Sup35NM monomers, which specifically leads to the formation of highly infectious strain conformations with more limited amyloid cores. Thus, transient non-native interactions in the initial nucleus are pivotal in determining the diversity of amyloid conformations and resulting prion strain phenotypes.
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Acknowledgements
We thank Y. Sakamaki and T. Akagi for help with the EM experiments and S. Akiyama for discussion of the SAXS measurements. Y.O. is supported by a Japan Society for the Promotion of Science (JSPS) postdoctoral fellowship. Funding was provided by the JST-PRESTO, the Ministry of Education, Culture, Sports, Science and Technology (MEXT), the Ministry of Health, Labour and Welfare (MHLW) and The Uehara Memorial Foundation (M.T.) and by the Howard Hughes Medical Institute and the US National Institutes of Health (J.S.W.).
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M.T. and Y.O. designed the experiments. Y.O. performed most experiments, K.I. contributed to setup of the SAXS experiments and B.H.T. helped the NMR experiments. B.H.T. and J.S.W. provided new reagents. Y.O., J.S.W. and M.T. analyzed and discussed the data and wrote the paper.
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Ohhashi, Y., Ito, K., Toyama, B. et al. Differences in prion strain conformations result from non-native interactions in a nucleus. Nat Chem Biol 6, 225–230 (2010). https://doi.org/10.1038/nchembio.306
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DOI: https://doi.org/10.1038/nchembio.306
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