TY - JOUR

T1 - Direct proof of the amyloid nature of yeast prions [PSI+] and [PIN+] by the method of immunoprecipitation of native fibrils

AU - Sergeeva, Aleksandra V

AU - Belashova, Tatyana A

AU - Bondarev, Stanislav A

AU - Velizhanina, Marya E

AU - Barbitoff, Yury A

AU - Matveenko, Andrew G

AU - Valina, Anna A

AU - Simanova, Angelina L

AU - Zhouravleva, Galina A

AU - Galkin, Alexey P

N1 - Publisher Copyright: © 2021 The Author(s) 2021. Published by Oxford University Press on behalf of FEMS. All rights reserved.

PY - 2021/9/1

Y1 - 2021/9/1

N2 - Prions are proteins that can exist in several structurally and functionally distinct states, one or more of which is transmissible. Yeast proteins Sup35 and Rnq1 in prion state ([PSI+] and [PIN+], respectively) form oligomers and aggregates, which are transmitted from parents to offspring in a series of generations. Several pieces of indirect evidence indicate that these aggregates also possess amyloid properties, but their binding to amyloid-specific dyes has not been shown in vivo. Meanwhile, it is the specific binding to the Congo Red dye and birefringence in polarized light after such staining that is considered the gold standard for proving the amyloid properties of a protein. Here we used immunoprecipitation to extract native fibrils of the Sup35 and Rnq1 proteins from yeast strains with different prion status. These fibrils are detected by electron microscopy, stained with Congo Red and exhibit yellow-green birefringence after such staining. All these data show that the Sup35 and Rnq1 proteins in prion state form amyloid fibrils in vivo. The technology of fibrils extraction in combination with standard cytological methods can be used to identify new pathological and functional amyloids in any organism and to analyze the structural features of native amyloid fibrils.

AB - Prions are proteins that can exist in several structurally and functionally distinct states, one or more of which is transmissible. Yeast proteins Sup35 and Rnq1 in prion state ([PSI+] and [PIN+], respectively) form oligomers and aggregates, which are transmitted from parents to offspring in a series of generations. Several pieces of indirect evidence indicate that these aggregates also possess amyloid properties, but their binding to amyloid-specific dyes has not been shown in vivo. Meanwhile, it is the specific binding to the Congo Red dye and birefringence in polarized light after such staining that is considered the gold standard for proving the amyloid properties of a protein. Here we used immunoprecipitation to extract native fibrils of the Sup35 and Rnq1 proteins from yeast strains with different prion status. These fibrils are detected by electron microscopy, stained with Congo Red and exhibit yellow-green birefringence after such staining. All these data show that the Sup35 and Rnq1 proteins in prion state form amyloid fibrils in vivo. The technology of fibrils extraction in combination with standard cytological methods can be used to identify new pathological and functional amyloids in any organism and to analyze the structural features of native amyloid fibrils.

KW - Congo Red

KW - amyloid

KW - birefringence

KW - fibril immunoprecipitation

KW - prion

KW - yeast

UR - http://www.scopus.com/inward/record.url?scp=85116310148&partnerID=8YFLogxK

UR - https://www.mendeley.com/catalogue/6ad45a19-f6eb-32b7-ad9c-19129f6182a1/

U2 - 10.1093/femsyr/foab046

DO - 10.1093/femsyr/foab046

M3 - Article

C2 - 34463335

VL - 21

JO - FEMS Yeast Research

JF - FEMS Yeast Research

SN - 1567-1356

IS - 6

M1 - foab046

ER -