TY - JOUR

T1 - Amyloid properties of the yeast cell wall protein Toh1 and its interaction with prion proteins Rnq1 and Sup35

AU - Sergeeva, A. V.

AU - Sopova, J. V.

AU - Belashova, T. A.

AU - Siniukova, V. A.

AU - Chirinskaite, A. V.

AU - Galkin, A. P.

AU - Zadorsky, S. P.

N1 - Funding Information: This work was supported by state program # 0112-2016-0015 to A.P.G., Saint-Petersburg University under research Grant № 1.40.505.2017 to A.P.G., RFBR under Grant № 18-34-00420 to A.V.S and RSF under Grant № 14-50-00069. The authors acknowledge St. Petersburg State University for opportunity to use facilities of the Research Resource Center “Development of Molecular and Cell Technologies” and the Resource Center “CHROMAS” of SPbSU. The authors are grateful to D.V. Kachkin for the assistance with FRET method. Funding Information: This work was supported by the Russian Federation State Program [0112-2016-0015]; SPbSU [1.40.505.2017];Russian Foundation for Basic Research [18-34-00420]; Russian Science Foundation [14-50-00069]. Publisher Copyright: © 2019, © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. Copyright: Copyright 2019 Elsevier B.V., All rights reserved.

PY - 2019/1/1

Y1 - 2019/1/1

N2 - Amyloids are non-branching fibrils that are composed of stacked monomers stabilized by intermolecular β-sheets. Some amyloids are associated with incurable diseases, whereas others, functional amyloids, regulate different vital processes. The prevalence and significance of functional amyloids in wildlife are still poorly understood. In recent years, by applying new approach of large-scale proteome screening, a number of novel candidate amyloids were identified in the yeast Saccharomyces cerevisiae, many of which are localized in the yeast cell wall. In this work, we showed that one of these proteins, Toh1, possess amyloid properties. The Toh1-YFP hybrid protein forms detergent-resistant aggregates in the yeast cells while being expressed under its own P TOH1 or inducible P CUP1 promoter. Using bacterial system for generation of extracellular amyloid aggregates C-DAG, we demonstrated that the N-terminal Toh1 fragment, containing amyloidogenic regions predicted in silico, binds Congo Red dye, manifests ‘apple-green’ birefringence when examined between crossed polarizers, and forms amyloid-like fibrillar aggregates visualized by TEM. We have established that the Toh1(20–365)-YFP hybrid protein fluorescent aggregates are co-localized with a high frequency with Rnq1C-CFP and Sup35NM-CFP aggregates in the yeast cells containing [PIN + ] and [PSI + ] prions, and physical interaction of these aggregated proteins was confirmed by FRET. This is one of a few known cases of physical interaction of non-Q/N-rich amyloid-like protein and Q/N-rich amyloids, suggesting that interaction of different amyloid proteins may be determined not only by similarity of their primary structures but also by similarity of their secondary structures and of conformational folds.

AB - Amyloids are non-branching fibrils that are composed of stacked monomers stabilized by intermolecular β-sheets. Some amyloids are associated with incurable diseases, whereas others, functional amyloids, regulate different vital processes. The prevalence and significance of functional amyloids in wildlife are still poorly understood. In recent years, by applying new approach of large-scale proteome screening, a number of novel candidate amyloids were identified in the yeast Saccharomyces cerevisiae, many of which are localized in the yeast cell wall. In this work, we showed that one of these proteins, Toh1, possess amyloid properties. The Toh1-YFP hybrid protein forms detergent-resistant aggregates in the yeast cells while being expressed under its own P TOH1 or inducible P CUP1 promoter. Using bacterial system for generation of extracellular amyloid aggregates C-DAG, we demonstrated that the N-terminal Toh1 fragment, containing amyloidogenic regions predicted in silico, binds Congo Red dye, manifests ‘apple-green’ birefringence when examined between crossed polarizers, and forms amyloid-like fibrillar aggregates visualized by TEM. We have established that the Toh1(20–365)-YFP hybrid protein fluorescent aggregates are co-localized with a high frequency with Rnq1C-CFP and Sup35NM-CFP aggregates in the yeast cells containing [PIN + ] and [PSI + ] prions, and physical interaction of these aggregated proteins was confirmed by FRET. This is one of a few known cases of physical interaction of non-Q/N-rich amyloid-like protein and Q/N-rich amyloids, suggesting that interaction of different amyloid proteins may be determined not only by similarity of their primary structures but also by similarity of their secondary structures and of conformational folds.

KW - амилоиды

KW - функциональные амилоиды

KW - дрожжи

KW - клеточная стенка

KW - Прионы дрожжей

UR - http://www.scopus.com/inward/record.url?scp=85059298582&partnerID=8YFLogxK

U2 - 10.1080/19336896.2018.1558763

DO - 10.1080/19336896.2018.1558763

M3 - Article

C2 - 30558459

AN - SCOPUS:85059298582

VL - 13

SP - 21

EP - 32

JO - Prion

JF - Prion

SN - 1933-6896

IS - 1

ER -