De novo generation of a PrPSc-like conformation in living cells.
Conformational conversion of the cellular PrPC protein to PrPSc is a central aspect of the prion diseases, but how PrP initially converts to this conformation remains a mystery. Here we show that PrP expressed in the yeast cytoplasm, instead of the endoplasmic reticulum, acquires the characteristics of PrPSc, namely detergent insolubility and a distinct pattern of protease resistance. Neuroblastoma cells cultured under reducing, glycosylation-inhibiting conditions produce PrP with the same characteristics. We therefore describe what is, to our knowledge, the first conversion of full-length PrP in a heterologous system, show the importance of reducing and deglycosylation conditions in PrP conformational transitions, and suggest a model for initiating events in sporadic and inherited prion diseases.
Ma
J
J
Howard Hughes Medical Institute, Department of Molecular Genetics and Cell Biology, University of Chicago, 5841 S. Maryland Avenue MC1028, Chicago, Illinois 60637, USA.
Lindquist
S
S
D000818:Animals; D003593:Cytoplasm / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D019286:Endopeptidase K / Q000378:metabolism; D015870:Gene Expression; D051379:Mice; D009447:Neuroblastoma; D018620:PrPSc Proteins / Q000096:biosynthesis* / Q000737:chemistry; D011328:Prions / Q000235:genetics; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae; D014407:Tumor Cells, Cultured; D020798:Two-Hybrid System Techniques
D018620:PrPSc Proteins; D011328:Prions; D019286:Endopeptidase K