Dissection and Design of Yeast Prions

Many proteins can misfold into β-sheet-rich, self-seeding polymers (amyloids). Prions are exceptional among such aggregates in that they are also infectious. In fungi, prions are not pathogenic but rather act as epigenetic regulators of cell physiology, providing a powerful model for studying the mechanism of prion replication. We used prion-forming domains from two budding yeast proteins (Sup35p and New1p) to examine the requirements for prion formation and inheritance. In both proteins, a glutamine/asparagine-rich (Q/N-rich) tract mediates sequence-specific aggregation, while an adjacent motif, the oligopeptide repeat, is required for the replication and stable inheritance of these aggregates. Our findings help to explain why although Q/N-rich proteins are relatively common, few form heritable aggregates: prion inheritance requires both an aggregation sequence responsible for self-seeded growth and an element that permits chaperone-dependent replication of the aggregate. Using this knowledge, we have designed novel artificial prions by fusing the replication element of Sup35p to aggregation-prone sequences from other proteins, including pathogenically expanded polyglutamine.

[1]  C. King,et al.  Supporting the structural basis of prion strains: induction and identification of [PSI] variants. , 2001, Journal of molecular biology.

[2]  Gary W. Jones,et al.  Amino acid residue 184 of yeast Hsp104 chaperone is critical for prion-curing by guanidine, prion propagation, and thermotolerance , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[3]  K. Wüthrich,et al.  Prion-inducing domain 2-114 of yeast Sup35 protein transforms in vitro into amyloid-like filaments. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Liebman,et al.  The Yeast [PSI +] Prion: Making Sense of Nonsense* , 1999, The Journal of Biological Chemistry.

[5]  Y. Chernoff,et al.  Yeast prion protein derivative defective in aggregate shearing and production of new ‘seeds’ , 2001, The EMBO journal.

[6]  H. Zoghbi,et al.  Glutamine repeats and neurodegeneration. , 2000, Annual review of neuroscience.

[7]  S. Lehmann.  [The prion protein]. , 2002, Journal de la Societe de biologie.

[8]  S. Liebman,et al.  Prions Affect the Appearance of Other Prions The Story of [PIN+] , 2001, Cell.

[9]  M. Tuite,et al.  Guanidine Hydrochloride Inhibits the Generation of Prion “Seeds” but Not Prion Protein Aggregation in Yeast , 2002, Molecular and Cellular Biology.

[10]  P. Lansbury,et al.  Amyloid diseases: abnormal protein aggregation in neurodegeneration. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  S. Paushkin,et al.  Propagation of the yeast prion‐like [psi+] determinant is mediated by oligomerization of the SUP35‐encoded polypeptide chain release factor. , 1996, The EMBO journal.

[12]  J. Weissman,et al.  Molecular Basis of a Yeast Prion Species Barrier , 2000, Cell.

[13]  M. Tuite,et al.  Analysis of the generation and segregation of propagons: entities that propagate the [PSI+] prion in yeast. , 2003, Genetics.

[14]  Jonathan S Weissman,et al.  Multiple Gln/Asn-Rich Prion Domains Confer Susceptibility to Induction of the Yeast [PSI+] Prion , 2001, Cell.

[15]  S. Lindquist,et al.  Rnq1: an epigenetic modifier of protein function in yeast. , 2000, Molecular cell.

[16]  M. Ter‐Avanesyan,et al.  Structure and Replication of Yeast Prions , 1998, Cell.

[17]  M. Ter‐Avanesyan,et al.  Prion properties of the Sup35 protein of yeast Pichia methanolica , 2000, The EMBO journal.

[18]  F. Lacroute.  Non-Mendelian Mutation Allowing Ureidosuccinic Acid Uptake in Yeast , 1971, Journal of bacteriology.

[19]  Robert A. Grothe,et al.  An amyloid-forming peptide from the yeast prion Sup35 reveals a dehydrated β-sheet structure for amyloid , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Y. Chernoff,et al.  Deletion analysis of the SUP35 gene of the yeast Saccharomyces cerevisiae reveals two non‐overlapping functional regions in the encoded protein , 1993, Molecular microbiology.

[21]  I. Herskowitz,et al.  Induction of Distinct [URE3] Yeast Prion Strains , 2001, Molecular and Cellular Biology.

[22]  S. Lindquist,et al.  Changes in the middle region of Sup35 profoundly alter the nature of epigenetic inheritance for the yeast prion [PSI+] , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[23]  D. Cheresh.  URE 3 ] as an Altered URE 2 Protein : Evidence for a Prion Analog in Saccharomyces cerevisiae , 2022 .

[24]  J R Glover,et al.  Support for the Prion Hypothesis for Inheritance of a Phenotypic Trait in Yeast , 1996, Science.

[25]  P. Zhou,et al.  The PNM2 mutation in the prion protein domain of SUP35 has distinct effects on different variants of the [PSI+] prion in yeast , 1999, Current Genetics.

[26]  S W Liebman,et al.  Role of the chaperone protein Hsp104 in propagation of the yeast prion-like factor [psi+]. , 1995, Science.

[27]  J. Weissman,et al.  Conformational diversity in a yeast prion dictates its seeding specificity , 2001, Nature.

[28]  Susan Lindquist,et al.  Prions as protein-based genetic elements. , 2002, Annual review of microbiology.

[29]  Yoshikazu Nakamura,et al.  Yeast [PSI+] "prions" that are crosstransmissible and susceptible beyond a species barrier through a quasi-prion state. , 2001, Molecular cell.

[30]  C. Nierras,et al.  The dominant PNM2- mutation which eliminates the psi factor of Saccharomyces cerevisiae is the result of a missense mutation in the SUP35 gene. , 1994, Genetics.

[31]  Stanley B. Prusiner,et al.  Nobel Lecture: Prions , 1998 .

[32]  J. Weissman,et al.  The utility of prions. , 2002, Developmental cell.

[33]  R. Wickner,et al.  Prion-Inducing Domain of Yeast Ure2p and Protease Resistance of Ure2p in Prion-Containing Cells , 1995, Science.

[34]  D. Kryndushkin,et al.  Yeast [PSI+] Prion Aggregates Are Formed by Small Sup35 Polymers Fragmented by Hsp104* , 2003, Journal of Biological Chemistry.

[35]  Y. Chernoff,et al.  Genetic and environmental factors affecting the de novo appearance of the [PSI+] prion in Saccharomyces cerevisiae. , 1997, Genetics.

[36]  M. Tuite,et al.  Mechanism of inhibition of Ψ+ prion determinant propagation by a mutation of the N‐terminus of the yeast Sup35 protein , 1998, The EMBO journal.

[37]  S. Lindquist,et al.  Aggregation of huntingtin in yeast varies with the length of the polyglutamine expansion and the expression of chaperone proteins. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[38]  R. Wickner,et al.  Prion domain initiation of amyloid formation in vitro from native Ure2p. , 1999, Science.

[39]  Yoshikazu Nakamura,et al.  Prion domain interaction responsible for species discrimination in yeast [PSI+] transmission , 2003, Genes to cells : devoted to molecular & cellular mechanisms.

[40]  S. Lindquist,et al.  Prion protein gene polymorphisms in Saccharomyces cerevisiae , 2003, Molecular microbiology.

[41]  S. Lindquist,et al.  Oligopeptide-repeat expansions modulate ‘protein-only’ inheritance in yeast , 1999, Nature.

[42]  B. Cox,et al.  The extrachromosomal control of nonsense suppression in yeast: An analysis of the elimination of [psi+] in the presence of a nuclear gene PNM- , 1977, Molecular and General Genetics MGG.

[43]  S. Lindquist,et al.  Rnq 1 : An Epigenetic Modifier of Protein , 2000 .

[44]  R. Wickner,et al.  [URE3] as an altered URE2 protein: evidence for a prion analog in Saccharomyces cerevisiae. , 1994, Science.

[45]  R. Sikorski,et al.  A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. , 1989, Genetics.

[46]  Y. Chernoff,et al.  Huntingtin toxicity in yeast model depends on polyglutamine aggregation mediated by a prion-like protein Rnq1 , 2002, The Journal of cell biology.

[47]  Y. Chernoff,et al.  Evolutionary conservation of prion‐forming abilities of the yeast Sup35 protein , 2000, Molecular microbiology.

[48]  C. Dobson.  Protein misfolding, evolution and disease. , 1999, Trends in biochemical sciences.

[49]  S. Lindquist,et al.  Self-Seeded Fibers Formed by Sup35, the Protein Determinant of [PSI +], a Heritable Prion-like Factor of S. cerevisiae , 1997, Cell.

[50]  J. Weissman,et al.  A Critical Role for Amino-Terminal Glutamine/Asparagine Repeats in the Formation and Propagation of a Yeast Prion , 1998, Cell.

[51]  M. Tuite,et al.  Oligopeptide repeats in the yeast protein Sup35p stabilize intermolecular prion interactions , 2001, The EMBO journal.

[52]  J. Weissman,et al.  A census of glutamine/asparagine-rich regions: implications for their conserved function and the prediction of novel prions. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[53]  Yoshikazu Nakamura,et al.  [PHI+], a novel Sup35‐prion variant propagated with non‐Gln/Asn oligopeptide repeats in the absence of the chaperone protein Hsp104 , 2003, Genes to cells : devoted to molecular & cellular mechanisms.

[54]  F. Sherman.  Getting started with yeast. , 1991, Methods in enzymology.

[55]  Johannes Buchner,et al.  Protein folding handbook , 2005 .

[56]  B. Cox,et al.  Ψ, A cytoplasmic suppressor of super-suppressor in yeast , 1965, Heredity.

[57]  F E Cohen,et al.  Pathologic conformations of prion proteins. , 1998, Annual review of biochemistry.

[58]  M. Arisawa,et al.  Cloning of the Candida glabrata TRP1 and HIS3 genes, and construction of their disruptant strains by sequential integrative transformation. , 1995, Gene.

[59]  Sean R. Collins,et al.  Generation of prion transmission barriers by mutational control of amyloid conformations , 2003, Nature.