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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Dobson, C. M. The structural basis of protein folding and its links with human disease. Phil. Trans. R. Soc. Lond. B 356, 133–145 (2001)
DePace, A. H. & Weissman, J. S. Origins and kinetic consequences of diversity in Sup35 yeast prion fibers. Nature Struct. Biol. 9, 389–396 (2002)
Baxa, U., Speransky, V., Steven, A. C. & Wickner, R. B. Mechanism of inactivation on prion conversion of the Saccharomyces cerevisiae Ure2 protein. Proc. Natl Acad. Sci. USA 99, 5253–5260 (2002)
Glover, J. R. et al. Self-seeded fibers formed by Sup35, the protein determinant of [PSI+], a heritable prion-like factor of S. cerevisiae. Cell 89, 811–819 (1997)
Kad, N. M., Thomson, N. H., Smith, D. P., Smith, D. A. & Radford, S. E. β2-Microglobulin and its deamidated variant, N17D form amyloid fibrils with a range of morphologies in vitro. J. Mol. Biol. 313, 559–571 (2001)
Lashuel, H. A., Hartley, D., Petre, B. M., Walz, T. & Lansbury, P. T. Jr Neurodegenerative disease: amyloid pores from pathogenic mutations. Nature 418, 291 (2002)
Collinge, J. Prion diseases of humans and animals: their causes and molecular basis. Annu. Rev. Neurosci. 24, 519–550 (2001)
Santoso, A., Chien, P., Osherovich, L. Z. & Weissman, J. S. Molecular basis of a yeast prion species barrier. Cell 100, 277–288 (2000)
Come, J. H., Fraser, P. E. & Lansbury, P. T. Jr A kinetic model for amyloid formation in the prion diseases: importance of seeding. Proc. Natl Acad. Sci. USA 90, 5959–5963 (1993)
Chernoff, Y. O. et al. Evolutionary conservation of prion-forming abilities of the yeast Sup35 protein. Mol. Microbiol. 35, 865–876 (2000)
Kushnirov, V. V., Kochneva-Pervukhova, N. V., Chechenova, M. B., Frolova, N. S. & Ter-Avanesyan, M. D. Prion properties of the Sup35 protein of yeast Pichia methanolica. EMBO J. 19, 324–331 (2000)
Nakayashiki, T., Ebihara, K., Bannai, H. & Nakamura, Y. Yeast [PSI+] “prions” that are crosstransmissible and susceptible beyond a species barrier through a quasi-prion state. Mol. Cell 7, 1121–1130 (2001)
Uptain, S. M. & Lindquist, S. Prions as protein-based genetic elements. Annu. Rev. Microbiol. 56, 703–741 (2002)
King, C. Y. et al. Prion-inducing domain 2-114 of yeast Sup35 protein transforms in vitro into amyloid-like filaments. Proc. Natl Acad. Sci. USA 94, 6618–6622 (1997)
Derkatch, I. L., Chernoff, Y. O., Kushnirov, V. V., Inge-Vechtomov, S. G. & Liebman, S. W. Genesis and variability of [PSI] prion factors in Saccharomyces cerevisiae. Genetics 144, 1375–1386 (1996)
Kushnirov, V. V., Kryndushkin, D. S., Boguta, M., Smirnov, V. N. & Ter-Avanesyan, M. D. Chaperones that cure yeast artificial [PSI+] and their prion-specific effects. Curr. Biol. 10, 1443–1446 (2000)
Zhou, P. et al. The yeast non-Mendelian factor [ETA+] is a variant of [PSI+], a prion-like form of release factor eRF3. EMBO J. 18, 1182–1191 (1999)
Kochneva-Pervukhova, N. V. et al. [PSI+] prion generation in yeast: characterization of the ‘strain’ difference. Yeast 18, 489–497 (2001)
Uptain, S. M., Sawicki, G. J., Caughey, B. & Lindquist, S. Strains of [PSI+] are distinguished by their efficiencies of prion-mediated conformational conversion. EMBO J. 20, 6236–6245 (2001)
King, C. Y. Supporting the structural basis of prion strains: induction and identification of [PSI] variants. J. Mol. Biol. 307, 1247–1260 (2001)
Chien, P. & Weissman, J. S. Conformational diversity in a yeast prion dictates its seeding specificity. Nature 410, 223–227 (2001)
DePace, A. H., Santoso, A., Hillner, P. & Weissman, J. S. A critical role for amino-terminal glutamine/asparagine repeats in the formation and propagation of a yeast prion. Cell 93, 1241–1252 (1998)
Barron, R. M. et al. Changing a single amino acid in the N-terminus of murine PrP alters TSE incubation time across three species barriers. EMBO J. 20, 5070–5078 (2001)
Mastrianni, J. A. et al. Inherited prion disease caused by the V210I mutation: transmission to transgenic mice. Neurology 57, 2198–2205 (2001)
Peretz, D. et al. A change in the conformation of prions accompanies the emergence of a new prion strain. Neuron 34, 921–932 (2002)
Bartz, J. C., Bessen, R. A., McKenzie, D., Marsh, R. F. & Aiken, J. M. Adaptation and selection of prion protein strain conformations following interspecies transmission of transmissible mink encephalopathy. J. Virol. 74, 5542–5547 (2000)
Hill, A. F., Antoniou, M. & Collinge, J. Protease-resistant prion protein produced in vitro lacks detectable infectivity. J. Gen. Virol. 80, 11–14 (1999)
Baskakov, I. V., Legname, G., Baldwin, M. A., Prusiner, S. B. & Cohen, F. E. Pathway complexity of prion protein assembly into amyloid. J. Biol. Chem. 277, 21140–21148 (2002)
Post, K., Brown, D. R., Groschup, M., Kretzschmar, H. A. & Riesner, D. Neurotoxicity but not infectivity of prion proteins can be induced reversibly in vitro. Arch. Virol. Suppl. 16, 265–273 (2000)
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.
The authors declare that they have no competing financial interests.
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