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Differences in prion strain conformations result from non-native interactions in a nucleus

Nature Chemical Biology volume 6, pages 225–230 (2010)Cite this article

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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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Figure 1: Sup35NM forms oligomers in a temperature-dependent, reversible manner.
Figure 2: Low-temperature Sup35NM oligomers are on the pathway to formation of Sc4 amyloid fibers.
Figure 3: Distinct amino acid regions in the prion domain of Sup35NM are involved in nucleation and amyloid growth.
Figure 4: Non-native interactions outside of the Sc4 amyloid core drive oligomer formation of Sup35NM.
Figure 5: Global relationships between Sup35NM monomer, initial nucleus, amyloid conformation and prion strain phenotype.

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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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Authors and Affiliations

  1. Tanaka Research Unit, RIKEN Brain Science Institute, Wako, Saitama, Japan

    Yumiko Ohhashi & Motomasa Tanaka

  2. RIKEN Spring-8 Center, Sayo, Hyogo, Japan

    Kazuki Ito

  3. Department of Cellular and Molecular Pharmacology, Howard Hughes Medical Institute, University of California, San Francisco, San Francisco, California, USA

    Brandon H Toyama & Jonathan S Weissman

  4. Precursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Saitama, Japan

    Motomasa Tanaka

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  1. Yumiko Ohhashi
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Contributions

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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Correspondence to Motomasa Tanaka.

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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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