Biochemical, Cell Biological, and Genetic Assays to Analyze Amyloid and Prion Aggregation in Yeast

Alberti, Simon; Halfmann, Randal; Lindquist, Susan

doi:10.1016/s0076-6879(10)70030-6

Cited by 78 publications

(102 citation statements)

References 60 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This method can distinguish between SDS-insoluble amyloids and SDS-soluble assemblies (34). In Q103-GFP-and N47-GFP-expressing cells, the majority of the GFP fusion protein was found in the SDS-soluble fraction (Fig.…”

Section: Aggregation-prone Prion-like Proteins Do Not Aggregate In Thementioning

confidence: 99%

See 1 more Smart Citation

Dictyostelium discoideum has a highly Q/N-rich proteome and shows an unusual resilience to protein aggregation

Malinovska

Palm

Gibson

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

Many protein-misfolding diseases are caused by proteins carrying prion-like domains. These proteins show sequence similarity to yeast prion proteins, which can interconvert between an intrinsically disordered and an aggregated prion state. The natural presence of prions in yeast has provided important insight into disease mechanisms and cellular proteostasis. However, little is known about prions in other organisms, and it is not yet clear whether the findings in yeast can be generalized. Using bioinformatics tools, we show that Dictyostelium discoideum has the highest content of prion-like proteins of all organisms investigated to date, suggesting that its proteome has a high overall aggregation propensity. To study mechanisms regulating these proteins, we analyze the behavior of several well-characterized prion-like proteins, such as an expanded version of human huntingtin exon 1 (Q103) and the prion domain of the yeast prion protein Sup35 (NM), in D. discoideum. We find that these proteins remain soluble and are innocuous to D. discoideum, in contrast to other organisms, where they form cytotoxic cytosolic aggregates. However, when exposed to conditions that compromise molecular chaperones, these proteins aggregate and become cytotoxic. We show that the disaggregase Hsp101, a molecular chaperone of the Hsp100 family, dissolves heat-induced aggregates and promotes thermotolerance. Furthermore, prion-like proteins accumulate in the nucleus, where they are targeted by the ubiquitin-proteasome system. Our data suggest that D. discoideum has undergone specific adaptations that increase the proteostatic capacity of this organism and allow for an efficient regulation of its prion-like proteome. molecular chaperones | proteostasis | Dictyostelium discoideum | protein aggregation | prion

show abstract

Section: Aggregation-prone Prion-like Proteins Do Not Aggregate In Thementioning

confidence: 99%

“…These so-called disaggregases are essential for thermotolerance (34,(58)(59)(60). We searched the proteome of D. discoideum for orthologs of Hsp100s proteins and found a potential candidate protein: Hsp101 (Fig.…”

Section: Heat-induced Aggregation Of Prion-like Proteins Is Reversed mentioning

confidence: 99%

Dictyostelium discoideum has a highly Q/N-rich proteome and shows an unusual resilience to protein aggregation

Malinovska

Palm

Gibson

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Several studies have employed fusions of known or suspected PrDs to GFP, Sup35C, Ure2C, or glucocorticoid receptor reporters (Patino et al 1996;Li and Lindquist 2000;Sondheimer and Lindquist 2000;Alberti et al 2009Alberti et al , 2010. These chimera often, but not always, behave as prions.…”

Section: Artificial and Heterologous Prions That Can Propagate In Yeastmentioning

confidence: 99%

Prions in Yeast

2012

View full text Add to dashboard Cite

The concept of a prion as an infectious self-propagating protein isoform was initially proposed to explain certain mammalian diseases. It is now clear that yeast also has heritable elements transmitted via protein. Indeed, the "protein only" model of prion transmission was first proven using a yeast prion. Typically, known prions are ordered cross-b aggregates (amyloids). Recently, there has been an explosion in the number of recognized prions in yeast. Yeast continues to lead the way in understanding cellular control of prion propagation, prion structure, mechanisms of de novo prion formation, specificity of prion transmission, and the biological roles of prions. This review summarizes what has been learned from yeast prions.

show abstract

“…Semi-denaturing detergent agarose gel electrophoresis SDD-AGE was performed as described previously (Alberti et al, 2010) with slight modifications. Approximately 7.5×10 7 cells expressing HA-Jub6p were lysed in TMSN buffer [0.25 M sucrose, 10 mM Tris-HCl, pH 7.5, 10 mM MgCl 2 , 3 mM CaCl 2 , 0.016% NP-40, 1 mM PMSF, 1× complete protease inhibitor (cpi) cocktail (Roche), 0.1 μg/ml DAPI] using a Dounce homogenizer (15 strokes), and the nuclei were collected by centrifuging at 4500 g and 4°C for 5 min.…”

Section: Ectopic Expression Of Proteins In Vegetative Cellsmentioning

confidence: 99%

Heterochromatin aggregation during DNA elimination in Tetrahymena is facilitated by a prion-like protein

Kataoka

Mochizuki

2017

Journal of Cell Science

View full text Add to dashboard Cite

Regulated aggregations of prion and prion-like proteins play physiological roles in various biological processes. However, their structural roles in the nucleus are poorly understood. Here, we show that the prion-like protein Jub6p is involved in the regulation of chromatin structure in the ciliated protozoan Tetrahymena thermophila. Jub6p forms sodium dodecyl sulfate (SDS)-resistant aggregates when it is ectopically expressed in vegetative cells and binds to RNA in vitro. Jub6p is a heterochromatin component and is important for the formation of heterochromatin bodies during the process of programmed DNA elimination. We suggest that RNAprotein aggregates formed by Jub6p are an essential architectural component for the assembly of heterochromatin bodies.

show abstract

Biochemical, Cell Biological, and Genetic Assays to Analyze Amyloid and Prion Aggregation in Yeast

Abstract: ABSTRACT

Cited by 78 publications

References 60 publications

Dictyostelium discoideum has a highly Q/N-rich proteome and shows an unusual resilience to protein aggregation

Dictyostelium discoideum has a highly Q/N-rich proteome and shows an unusual resilience to protein aggregation

Prions in Yeast

Heterochromatin aggregation during DNA elimination in Tetrahymena is facilitated by a prion-like protein

Contact Info

Product

Resources

About