In vitro propagation of the prion-like state of yeast Sup35 protein.
The yeast cytoplasmically inherited genetic determinant [PSI+] is presumed to be a manifestation of the prion-like properties of the Sup35 protein (Sup35p). Here, cell-free conversion of Sup35p from [psi-] cells (Sup35ppsi-) to the prion-like [PSI+]-specific form (Sup35pPSI+) was observed. The conversion reaction could be repeated for several consecutive cycles, thus modeling in vitro continuous [PSI+] propagation. Size fractionation of lysates of [PSI+] cells demonstrated that the converting activity was associated solely with Sup35pPSI+ aggregates, which agrees with the nucleation model for [PSI+] propagation. Sup35pPSI+ was purified and showed high conversion activity, thus confirming the prion hypothesis for Sup35p.
Paushkin
S V
SV
Institute of Experimental Cardiology, Cardiology Research Center, 3rd Cherepkovskaya Street 15A, Moscow 121552, Russia.
Kushnirov
V V
VV
Smirnov
V N
VN
Ter-Avanesyan
M D
MD
D010450:Endopeptidases / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008956:Models, Chemical; D010454:Peptide Termination Factors; D010641:Phenotype; D018621:PrPC Proteins / Q000737:chemistry; D018620:PrPSc Proteins / Q000737:chemistry; D011328:Prions / Q000737:chemistry*; D014176:Protein Biosynthesis; D011487:Protein Conformation; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000737:chemistry* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D012995:Solubility; D014170:Transformation, Genetic
D005656:Fungal Proteins; D010454:Peptide Termination Factors; D018621:PrPC Proteins; D018620:PrPSc Proteins; D011328:Prions; C068068:SUP35 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D010450:Endopeptidases