TY - JOUR
T1 - Live imaging of prions reveals nascent PrPSc in cellsurface, raft-associated amyloid strings and webs
AU - Rouvinski, Alexander
AU - Karniely, Sharon
AU - Kounin, Maria
AU - Moussa, Sanaa
AU - Goldberg, Miri D.
AU - Warburg, Gabriela
AU - Lyakhovetsky, Roman
AU - Papy-Garcia, Dulce
AU - Kutzsche, Janine
AU - Korth, Carsten
AU - Carlson, George A.
AU - Godsave, Susan F.
AU - Peters, Peter J.
AU - Luhr, Katarina
AU - Kristensson, Krister
AU - Taraboulos, Albert
PY - 2014/2/3
Y1 - 2014/2/3
N2 - Mammalian prions refold host glycosylphosphatidylinositol- anchored PrPC into β-sheet- rich PrPSc. PrPSc is rapidly truncated into a C-terminal PrP27-30 core that is stable for days in endolysosomes. The nature of cell-associated prions, their attachment to membranes and rafts, and their subcellular locations are poorly understood; live prion visualization has not previously been achieved. A key obstacle has been the inaccessibility of PrP27-30 epitopes. We overcame this hurdle by focusing on nascent full-length PrPSc rather than on its truncated PrP27-30 product. We show that N-terminal PrPSc epitopes are exposed in their physiological context and visualize, for the first time, PrPSc in living cells. PrPSc resides for hours in unexpected cell-surface, slow moving strings and webs, sheltered from endocytosis. Prion strings observed by light and scanning electron microscopy were thin, micrometerlong structures. They were firmly cell associated, resisted phosphatidylinositol-specific phospholipase C, aligned with raft markers, fluoresced with thioflavin, and were rapidly abolished by anti-prion glycans. Prion strings and webs are the first demonstration of membraneanchored PrPSc amyloids.
AB - Mammalian prions refold host glycosylphosphatidylinositol- anchored PrPC into β-sheet- rich PrPSc. PrPSc is rapidly truncated into a C-terminal PrP27-30 core that is stable for days in endolysosomes. The nature of cell-associated prions, their attachment to membranes and rafts, and their subcellular locations are poorly understood; live prion visualization has not previously been achieved. A key obstacle has been the inaccessibility of PrP27-30 epitopes. We overcame this hurdle by focusing on nascent full-length PrPSc rather than on its truncated PrP27-30 product. We show that N-terminal PrPSc epitopes are exposed in their physiological context and visualize, for the first time, PrPSc in living cells. PrPSc resides for hours in unexpected cell-surface, slow moving strings and webs, sheltered from endocytosis. Prion strings observed by light and scanning electron microscopy were thin, micrometerlong structures. They were firmly cell associated, resisted phosphatidylinositol-specific phospholipase C, aligned with raft markers, fluoresced with thioflavin, and were rapidly abolished by anti-prion glycans. Prion strings and webs are the first demonstration of membraneanchored PrPSc amyloids.
UR - http://www.scopus.com/inward/record.url?scp=84893454609&partnerID=8YFLogxK
U2 - https://doi.org/10.1083/jcb.201308028
DO - https://doi.org/10.1083/jcb.201308028
M3 - مقالة
C2 - 24493590
SN - 0021-9525
VL - 204
SP - 423
EP - 441
JO - Journal of Cell Biology
JF - Journal of Cell Biology
IS - 3
ER -