The misfolding of different cellular proteins into amyloid-like aggregates is associated with non- infectious neurodegenerative diseases including Alzheimer's, Huntington's and Parkinson's, as well as with the infectious prion diseases e.g. Mad Cow, Chronic Wasting in deer and Creutzfeldt-Jacob in humans. For each of these diseases, the associated protein aggregate ("seed') attracts its normal conformers to misfold and join the aggregate. Certain proteins in the simple eukaryote yeast, can likewise misfold into infectious amyloid aggregates, and these aggregates cause epigenetic variation. In this proposal, the power of yeast genetics and molecular biology is used to study how proteins misfold into amyloid-like aggregates and the consequences of this misfolding for the cell. The extensive similarity between yeast and human cells, which has enabled yeast models to make significant contributions in understanding human disease, implies that these studies will likely be relevant to misfolded aggregating proteins in humans. Since most human protein misfolding diseases occur without infection by any external seed, Aim I focuses on the molecular mechanisms surrounding spontaneous cellular amyloid formation. The questions addressed are: where do newly appearing prion aggregates first arise in cells, what other proteins are associated with them, and how do pre-existing prions enhance the de novo appearance of heterologous prions? Aim I also tests the hypothesis that Sla2, the yeast homolog of the mammalian huntingtin interacting protein, facilitates the ability of existing prions to cross-seed the de novo aggregation of heterologous prion proteins, by binding to both the seed and protein to be seeded, thereby placing them in close proximity. Interestingly, human and yeast prion proteins can each form multiple variants of amyloid aggregates that differ in structure and cause distinct phenotypes or disease pathologies, even though the amino acid sequences of the proteins are identical.
Aim II identifies proteins bound to, and/or required for, the propagation of several prions and their variants. In addition, solid-state NMR structures of two variants of the same prion will be determined with the help of collaborators. By comparing heterologous prions, as well as different variants of the same prion, factors likely to be common to the maintenance and infectivity of all prions and that should therefore provide useful drug targets, will be identified. While amyloid formation is associated with disease, the actual cause of pathology is unclear.
In Aim III, genetic and molecular studies of two prions that cause toxicity in yeast will help define the toxic species. Finally, yeast prions are important not only as a model for human disease, but also because they suggest an important new mechanism of genetic variation operating at the level of protein conformation rather than nucleic acids.
In Aim I V the fascinating question of whether prions can sometimes provide the host cell with an advantage is explored, along with the possibility that such advantageous prions may also exist in mammals.

Public Health Relevance

The power of yeast genetics and molecular biology will be used to study the misfolding of proteins into amyloid-like aggregates like those associated with several devastating neurodegenerative human diseases including Alzheimer's, Parkinson's, Huntington's and Creutzfeldt-Jacob's diseases. The extensive similarity between yeast and human cells, which has enabled yeast models to make significant contributions in understanding other human diseases, implies that the insights gained from these studies will be helpful when choosing drug-targets for the human diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM056350-16
Application #
8321542
Study Section
Membrane Biology and Protein Processing (MBPP)
Program Officer
Maas, Stefan
Project Start
1997-08-01
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2014-08-31
Support Year
16
Fiscal Year
2012
Total Cost
$423,488
Indirect Cost
$122,595
Name
University of Nevada Reno
Department
Biochemistry
Type
Schools of Medicine
DUNS #
146515460
City
Reno
State
NV
Country
United States
Zip Code
89557
Yang, Zi; Stone, David E; Liebman, Susan W (2014) Prion-promoted phosphorylation of heterologous amyloid is coupled with ubiquitin-proteasome system inhibition and toxicity. Mol Microbiol 93:1043-56
Yang, Zi; Hong, Joo Y; Derkatch, Irina L et al. (2013) Heterologous gln/asn-rich proteins impede the propagation of yeast prions by altering chaperone availability. PLoS Genet 9:e1003236
Derkatch, Irina L; Liebman, Susan W (2013) The story of stolen chaperones: how overexpression of Q/N proteins cures yeast prions. Prion 7:294-300
Sharma, Jaya; Liebman, Susan W (2013) Exploring the basis of [PIN(+)] variant differences in [PSI(+)] induction. J Mol Biol 425:3046-59
Mathur, Vidhu; Seuring, Carolin; Riek, Roland et al. (2012) Localization of HET-S to the cell periphery, not to [Het-s] aggregates, is associated with [Het-s]-HET-S toxicity. Mol Cell Biol 32:139-53
Manogaran, Anita L; Hong, Joo Y; Hufana, Joan et al. (2011) Prion formation and polyglutamine aggregation are controlled by two classes of genes. PLoS Genet 7:e1001386
Hong, Joo Y; Mathur, Vidhu; Liebman, Susan W (2011) A new colour assay for [URE3] prion in a genetic background used to score for the [PSI?] prion. Yeast 28:555-60
Chernova, Tatiana A; Romanyuk, Andrey V; Karpova, Tatiana S et al. (2011) Prion induction by the short-lived, stress-induced protein Lsb2 is regulated by ubiquitination and association with the actin cytoskeleton. Mol Cell 43:242-52
Mathur, Vidhu; Taneja, Vibha; Sun, Yidi et al. (2010) Analyzing the birth and propagation of two distinct prions, [PSI+] and [Het-s](y), in yeast. Mol Biol Cell 21:1449-61
Wagner, Jennifer K; Marquis, Kathleen A; Rudner, David Z (2009) SirA enforces diploidy by inhibiting the replication initiator DnaA during spore formation in Bacillus subtilis. Mol Microbiol 73:963-74

Showing the most recent 10 out of 36 publications