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Generation of prion transmission barriers by mutational control of amyloid conformations


Self-propagating β-sheet-rich protein aggregates are implicated in a wide range of protein-misfolding phenomena, including amyloid diseases and prion-based inheritance1. Two properties have emerged as common features of amyloids. Amyloid formation is ubiquitous: many unrelated proteins form such aggregates and even a single polypeptide can misfold into multiple forms2,3,4,5,6 — a process that is thought to underlie prion strain variation7. Despite this promiscuity, amyloid propagation can be highly sequence specific: amyloid fibres often fail to catalyse the aggregation of other amyloidogenic proteins8,9. In prions, this specificity leads to barriers that limit transmission between species7,8,10,11,12. Using the yeast prion [PSI+]13, we show in vitro that point mutations in Sup35p, the protein determinant of [PSI+], alter the range of ‘infectious’ conformations, which in turn changes amyloid seeding specificity. We generate a new transmission barrier in vivo by using these mutations to specifically disfavour subsets of prion strains. The ability of mutations to alter the conformations of amyloid states without preventing amyloid formation altogether provides a general mechanism for the generation of prion transmission barriers and may help to explain how mutations alter toxicity in conformational diseases.

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We thank H. Wille, J. Hood-DeGrenier and members of the Weissman and Lim lab for discussion and critical reading of the manuscript. P.C. and S.R.C. were supported by National Science Foundation Graduate Fellowships and the ARCS (Achievement Rewards for College Scientists) foundation (P.C.). A.H.D. was supported by a Howard Hughes Medical Institute predoctoral fellowship. Funding was also provided by Howard Hughes Medical Institute, The David and Lucile Packard Foundation and the National Institutes of Health.

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Correspondence to Jonathan S. Weissman.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Table 1: Expansions and explanations of abbreviations used in main text. (PDF 170 kb)

Supplementary Figure 1: Partial proteolysis of Ch fibres as generated by seed or temperature variation. (PDF 128 kb)

Supplementary Figure 2: Seeding specificity of Ch fibres generated at varying temperatures. (PDF 402 kb)

Supplementary Figure 3: Seeding specificity and thermal denaturation of ChQ15R fibres. Quantitation of thermal denaturation profiles of Figure 1 in main text. (PDF 321 kb)

Supplementary Figure 4: Colour coded sequence of Ch showing locations of mutants used in this work. (PDF 400 kb)

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Further reading

Figure 1: Conformation of chimaeric prion fibres is sensitive to polymerization conditions.
Figure 2: Mutations create a transmission barrier in vitro.
Figure 3: Mutations create a transmission barrier in vivo by shifting strain preference of chimaeric prion.
Figure 4: Effects of mutations on the natural S. cerevisiae Sup35 prion domain.


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