TY - JOUR

T1 - Mammalian amyloidogenic proteins promote prion nucleation in yeast

AU - Chandramowlishwaran, Pavithra

AU - Sun, Meng

AU - Casey, Kristin L

AU - Romanyuk, Andrey V

AU - Grizel, Anastasiya V

AU - Sopova, Julia V

AU - Rubel, Aleksandr A

AU - Nussbaum-Krammer, Carmen

AU - Vorberg, Ina M

AU - Chernoff, Yury O

N1 - © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

PY - 2018

Y1 - 2018

N2 - Fibrous cross-β aggregates (amyloids) and their transmissible forms (prions) cause diseases in mammals (including humans) and control heritable traits in yeast. Initial nucleation of a yeast prion by transiently overproduced prion-forming protein or its (typically, QN-rich) prion domain is efficient only in the presence of another aggregated (in most cases, QN-rich) protein. Here, we demonstrate that a fusion of the prion domain of yeast protein Sup35 to some non-QN-rich mammalian proteins, associated with amyloid diseases, promotes nucleation of Sup35 prions in the absence of preexisting aggregates. In contrast, both a fusion of the Sup35 prion domain to a multimeric non-amyloidogenic protein, and an expression of a mammalian amyloidogenic protein that is not fused to the Sup35 prion domain failed to promote prion nucleation, further indicating that physical linkage of a mammalian amyloidogenic protein to the prion domain of a yeast protein is required for the nucleation of a yeast prion. Biochemical and cytological approaches confirmed the nucleation of protein aggregates in the yeast cell. Sequence alterations antagonizing or enhancing amyloidogenicity of human β amyloid (Aβ, associated with Alzheimer disease) and mouse PrP (associated with prion diseases) respectively antagonized or enhanced nucleation of a yeast prion by these proteins. The yeast-based prion nucleation assay, developed in our work, can be employed for mutational dissection of amyloidogenic proteins. We anticipate that it will aid in the identification of chemicals that influence initial amyloid nucleation and in searching for new amyloidogenic proteins in a variety of proteomes.

AB - Fibrous cross-β aggregates (amyloids) and their transmissible forms (prions) cause diseases in mammals (including humans) and control heritable traits in yeast. Initial nucleation of a yeast prion by transiently overproduced prion-forming protein or its (typically, QN-rich) prion domain is efficient only in the presence of another aggregated (in most cases, QN-rich) protein. Here, we demonstrate that a fusion of the prion domain of yeast protein Sup35 to some non-QN-rich mammalian proteins, associated with amyloid diseases, promotes nucleation of Sup35 prions in the absence of preexisting aggregates. In contrast, both a fusion of the Sup35 prion domain to a multimeric non-amyloidogenic protein, and an expression of a mammalian amyloidogenic protein that is not fused to the Sup35 prion domain failed to promote prion nucleation, further indicating that physical linkage of a mammalian amyloidogenic protein to the prion domain of a yeast protein is required for the nucleation of a yeast prion. Biochemical and cytological approaches confirmed the nucleation of protein aggregates in the yeast cell. Sequence alterations antagonizing or enhancing amyloidogenicity of human β amyloid (Aβ, associated with Alzheimer disease) and mouse PrP (associated with prion diseases) respectively antagonized or enhanced nucleation of a yeast prion by these proteins. The yeast-based prion nucleation assay, developed in our work, can be employed for mutational dissection of amyloidogenic proteins. We anticipate that it will aid in the identification of chemicals that influence initial amyloid nucleation and in searching for new amyloidogenic proteins in a variety of proteomes.

KW - Journal Article

KW - Amyloid/metabolism

KW - Protein Aggregates

KW - Saccharomyces cerevisiae Proteins/chemistry

KW - Humans

KW - Protein Domains

KW - Peptide Fragments/metabolism

KW - Amyloid beta-Peptides/metabolism

KW - Peptide Termination Factors/chemistry

KW - ALZHEIMERS-DISEASE

KW - PSI+ PRION

KW - SACCHAROMYCES-CEREVISIAE

KW - ATOMIC-RESOLUTION STRUCTURE

KW - DE-NOVO APPEARANCE

KW - BETA-PROTEIN

KW - SECONDARY STRUCTURE

KW - ACTIN CYTOSKELETON

KW - TRANSLATION TERMINATION

KW - TRANSGENIC MICE

UR - http://www.scopus.com/inward/record.url?scp=85042937773&partnerID=8YFLogxK

UR - http://www.mendeley.com/research/mammalian-amyloidogenic-proteins-promote-prion-nucleation-yeast

U2 - 10.1074/jbc.M117.809004

DO - 10.1074/jbc.M117.809004

M3 - Article

C2 - 29330303

VL - 293

SP - 3436

EP - 3450

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 9

ER -