%0 Journal Article
%T Early Delivery of Misfolded PrP from ER to Lysosomes by Autophagy
%A Constanza J. Cortes
%A Kefeng Qin
%A Eric M. Norstrom
%A William N. Green
%A Vytautas P. Bindokas
%A James A. Mastrianni
%J International Journal of Cell Biology
%D 2013
%I Hindawi Publishing Corporation
%R 10.1155/2013/560421
%X Prion diseases are linked to the accumulation of a misfolded isoform (PrPSc) of prion protein (PrP). Evidence suggests that lysosomes are degradation endpoints and sites of the accumulation of PrPSc. We questioned whether lysosomes participate in the early quality control of newly generated misfolded PrP. We found PrP carrying the disease-associated T182A mutation (Mut-PrP) was delivered to lysosomes in a Golgi-independent manner. Time-lapse live cell imaging revealed early formation and uptake of GFP-tagged Mut-PrP aggregates into LysoTracker labeled vesicles. Compared with Wt-PrP, Mut-PrP expression was associated with an elevation in several markers of the autophagy-lysosomal pathway, and it extensively colocalized with the autophagosome-specific marker, LC3B. In autophagy deficient (ATG5£¿/£¿) mouse embryonic fibroblasts, or in normal cells treated with the autophagy-inhibitor 3-MA, Mut-PrP colocalization with lysosomes was reduced to a similar extent. Additionally, 3-MA selectively impaired the degradation of insoluble Mut-PrP, resulting in an increase in protease-resistant PrP, whereas the induction of autophagy by rapamycin reduced it. These findings suggest that autophagy might function as a quality control mechanism to limit the accumulation of misfolded PrP that normally leads to the generation of PrPSc. 1. Introduction Prion diseases, such as Creutzfeldt-Jakob disease (CJD) of humans and bovine spongiform encephalopathy (BSE) of cattle, are transmissible neurodegenerative disorders linked to the accumulation of a misfolded isoform (PrPSc) of the host-encoded glycophosphatidylinositol (GPI)-linked prion protein (PrPC) [1]. As a membrane protein, PrPC follows the secretory pathway to its destination on the outer leaflet of the plasma membrane where it ultimately follows the endocytic pathway for degradation in lysosomes. Mutations of the PrP gene linked to familial prion disease promote the misfolding of PrP that may delay its exit from the endoplasmic reticulum, leading to impaired delivery to the plasma membrane and an alternative pathway for degradation. Autophagy is an evolutionarily conserved lysosomal degradation pathway usually activated under low nutrient conditions which acts to sequester and deliver cytoplasmic material, including organelles, toxic metabolites, or intracellular pathogens, to the lysosome for degradation and/or recycling [2]. This process is highly regulated by a series of autophagy-related gene products or Atg proteins [3, 4]. Key proteins include Atg6 and its mammalian homolog Beclin-1, which participate in the
%U http://www.hindawi.com/journals/ijcb/2013/560421/