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Correlation of structural elements and infectivity of the HET-s prion

Nature volume 435, pages 844–848 (2005)Cite this article

Abstract

Prions are believed to be infectious, self-propagating polymers of otherwise soluble, host-encoded proteins1,2. This concept is now strongly supported by the recent findings that amyloid fibrils of recombinant prion proteins from yeast3,4,5, Podospora anserina6 and mammals7 can induce prion phenotypes in the corresponding hosts. However, the structural basis of prion infectivity remains largely elusive because acquisition of atomic resolution structural properties of amyloid fibrils represents a largely unsolved technical challenge. HET-s, the prion protein of P. anserina, contains a carboxy-terminal prion domain comprising residues 218–289. Amyloid fibrils of HET-s(218–289) are necessary and sufficient for the induction and propagation of prion infectivity6. Here, we have used fluorescence studies, quenched hydrogen exchange NMR and solid-state NMR to determine the sequence-specific positions of amyloid fibril secondary structure elements of HET-s(218–289). This approach revealed four β-strands constituted by two pseudo-repeat sequences, each forming a β-strand-turn-β-strand motif. By using a structure-based mutagenesis approach, we show that this conformation is the functional and infectious entity of the HET-s prion. These results correlate distinct structural elements with prion infectivity.

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Figure 1: Sequence-specific determination of regular secondary structure and its topology in HET-s(218-289) fibrils.
Figure 2: The proposed fold of the infectious conformation of HET-s(218–289) amyloid fibrils.
Figure 3: In vivo prion formation of HET-s proline and deletion mutants.

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Acknowledgements

R.R. is a Pew scholar. This research was supported in part by grants from the National Institute of Health, the US Army, the ETH Zurich, the Swiss National Science Foundation, the CNRS and the French Ministry of Research.

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Author notes

  1. Christiane Ritter, Marie-Lise Maddelein and Ansgar B. Siemer: *These authors contributed equally to this work

Authors and Affiliations

  1. The Salk Institute, 10010 North Torrey Pines Road, California, 92037, La Jolla, USA

    Christiane Ritter, Thorsten Lührs & Roland Riek

  2. Laboratoire de Génétique Moléculaire des Champignons, Institut de Biochimie et de Génétique Cellulaires, Unité Mixte de Recherche 5095, Centre national de la Recherche Scientifique Université de Bordeaux 2, 33077, Bordeaux Cedex, France

    Marie-Lise Maddelein & Sven J. Saupe

  3. ETH Zurich, Physical Chemistry, ETH Honggerberg, 8093, Zurich, Switzerland

    Ansgar B. Siemer, Matthias Ernst & Beat H. Meier

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Correspondence to Roland Riek.

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Supplementary information

Supplementary Notes

Contains Supplementary Figures S1-S5 (additional solid and liquid state NMR data, sequence alignments and EM pictures), legends to accompany the figures and Supplementary Tables S1 and S2 giving details of the HET-s infectivity and function assays. (PDF 2848 kb)

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Ritter, C., Maddelein, ML., Siemer, A. et al. Correlation of structural elements and infectivity of the HET-s prion. Nature 435, 844–848 (2005). https://doi.org/10.1038/nature03793

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