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J Biol Chem. 2018 Jul 20;293(29):11424-11432. doi: 10.1074/jbc.RA117.001628. Epub 2018 May 31.

A hydrophobic low-complexity region regulates aggregation of the yeast pyruvate kinase Cdc19 into amyloid-like aggregates in vitro.

Author information

1
From the Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering and.
2
the Department of Biology, Institute for Biochemistry, ETH Zurich, 8093 Zurich, Switzerland.
3
From the Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering and paolo.arosio@chem.ethz.ch.

Abstract

Cells form stress granules (SGs) upon stress stimuli to protect sensitive proteins and RNA from degradation. In the yeast Saccharomyces cerevisiae, specific stresses such as nutrient starvation and heat-shock trigger recruitment of the yeast pyruvate kinase Cdc19 into SGs. This RNA-binding protein was shown to form amyloid-like aggregates that are physiologically reversible and essential for cell cycle restart after stress. Cellular Cdc19 exists in an equilibrium between a homotetramer and monomer state. Here, we show that Cdc19 aggregation in vitro is governed by protein quaternary structure, and we investigate the physical-chemical basis of Cdc19's assembly properties. Equilibrium shift toward the monomer state exposes a hydrophobic low-complexity region (LCR), which is prone to induce intermolecular interactions with surrounding proteins. We further demonstrate that hydrophobic/hydrophilic interfaces can trigger Cdc19 aggregation in vitro Moreover, we performed in vitro biophysical analyses to compare Cdc19 aggregates with fibrils produced by two known dysfunctional amyloidogenic peptides. We show that the Cdc19 aggregates share several structural features with pathological amyloids formed by human insulin and the Alzheimer's disease-associated Aβ42 peptide, particularly secondary β-sheet structure, thermodynamic stability, and staining by the thioflavin T dye. However, Cdc19 aggregates could not seed aggregation. These results indicate that Cdc19 adopts an amyloid-like structure in vitro that is regulated by the exposure of a hydrophobic LCR in its monomeric form. Together, our results highlight striking structural similarities between functional and dysfunctional amyloids and reveal the crucial role of hydrophobic/hydrophilic interfaces in regulating Cdc19 aggregation.

KEYWORDS:

Aβ42; amyloidogenic protein; cell compartmentalization; functional amyloids; hydrophobic-hydrophilic interface; protein aggregation; protein self-assembly; protein-nucleic acid interaction; protein-protein interaction; stress granule

PMID:
29853641
PMCID:
PMC6065187
DOI:
10.1074/jbc.RA117.001628
[Indexed for MEDLINE]
Free PMC Article

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