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Rapid cell-surface prion protein conversion revealed using a novel cell system

By R. Goold, S. Rabbanian, L. Sutton, R. Andre, P. Arora, J. Moonga, A.R. Clarke, G. Schiavo, P. Jat, J. Collinge and S.J. Tabrizi

Abstract

Prion diseases are fatal neurodegenerative disorders with unique transmissible properties. The infectious and pathological agent is thought to be a misfolded conformer of the prion protein. Little is known about the initial events in prion infection because the infecting prion source has been immunologically indistinguishable from normal cellular prion protein (PrPC). Here we develop a unique cell system in which epitope-tagged PrPC is expressed in a PrP knockdown (KD) neuroblastoma cell line. The tagged PrPC, when expressed in our PrP-KD cells, supports prion replication with the production of bona fide epitope-tagged infectious misfolded PrP (PrPSc). Using this epitope-tagged PrPSc, we study the earliest events in cellular prion infection and PrP misfolding. We show that prion infection of cells is extremely rapid occurring within 1 min of prion exposure, and we demonstrate that the plasma membrane is the primary site of prion conversion

Topics: Article
Publisher: Nature Publishing Group
OAI identifier: oai:pubmedcentral.nih.gov:3104518
Provided by: PubMed Central

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Citations

  1. (1988). A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.
  2. (2004). A large-scale RNAi screen in human cells identifies new components of the p53 pathway.
  3. (2003). A quantitative, highly sensitive cell-based infectivity assay for mouse scrapie prions.
  4. (1992). Caveolin, a protein component of caveolae membrane coats.
  5. (2009). Cells expressing anchorless prion protein are resistant to scrapie infection.
  6. (2010). Cellular factors implicated in prion replication.
  7. (2009). Cholesterol synthesis inhibitor U18666A and the role of sterol metabolism and trafficking in numerous pathophysiological processes.
  8. (2002). Conversion of PrP to a self-perpetuating PrPSc-like conformation in the cytosol.
  9. (2000). Cultured cell sublines highly susceptible to prion infection.
  10. (2008). Detection and characterization of proteinase K-sensitive disease-related prion protein with thermolysin.
  11. (2005). Disease-related prion protein forms aggresomes in neuronal cells leading to caspase activation and apoptosis.
  12. (2006). Dynasore, a cell-permeable inhibitor of dynamin.
  13. (1992). Evidence for synthesis of scrapie prion proteins in the endocytic pathway.
  14. (2002). Filipin prevents pathological prion protein accumulation by reducing endocytosis and inducing cellular PrP release.
  15. (2007). Formation of native prions from minimal components in vitro.
  16. (1999). Functionally different GPI proteins are organized in different domains on the neuronal surface.
  17. (2010). Generating a prion with bacterially expressed recombinant prion protein.
  18. (2009). Glypican-1 mediates both prion protein lipid raft association and disease isoform formation.
  19. (1994). Heterologous PrP molecules interfere with accumulation of protease-resistant PrP in scrapieinfected murine neuroblastoma cells.
  20. (2009). Identification of an intracellular site of prion conversion.
  21. (2008). Immunolocalisation of PrP(Sc) in scrapie-infected N2a mouse neuroblastoma cells by light and electron microscopy.
  22. (1984). Initial steps in receptormediated endocytosis. The influence of temperature on the shape and distribution of plasma membrane clathrin-coated pits in cultured mammalian cells.
  23. (2001). Interactions between prion protein isoforms: the kiss of death?
  24. (2000). Involvement of p38 mitogen-activated protein kinase signaling pathway in osteoclastogenesis mediated by receptor activator of NF-kappa
  25. (2008). Mouse neuroblastoma cells release prion infectivity associated with exosomal vesicles.
  26. (1997). N-terminally tagged prion protein supports prion propagation in transgenic mice.
  27. (2010). Neuronal low-density lipoprotein receptor-related protein 1 binds and endocytoses prion fibrils via receptor cluster 4.
  28. (1982). Novel proteinaceous infectious particles cause scrapie.
  29. Pathologic prion protein infects cells by lipid-raft dependent macropinocytosis.
  30. (2001). Prion diseases of humans and animals: their causes and molecular basis.
  31. (2006). Prion infection of mouse neurospheres.
  32. (1996). Prion protein (PrP) with amino-proximal deletions restoring susceptibility of PrP knockout mice to scrapie.
  33. (2006). Prions and their partners in crime.
  34. (2004). Prions can infect primary cultured neurons and astrocytes and promote neuronal cell death.
  35. (2009). Prions hijack tunnelling nanotubes for intercellular spread.
  36. (1994). Quantification of low density lipoprotein and transferrin endocytic sorting HEp2 cells using confocal microscopy.
  37. (2001). Scrapie prion protein accumulation by scrapie-infected neuroblastoma cells abrogated by exposure to a prion protein antibody.
  38. (1988). Scrapie-infected murine neuroblastoma cells produce protease-resistant prion proteins.
  39. (2008). Single treatment with RNAi against prion protein rescues early neuronal dysfunction and prolongs survival in mice with prion disease.
  40. (2000). Sites of prion protein accumulation in scrapie-infected mouse spleen revealed by immuno-electron microscopy.
  41. (2005). Species barriers in prion diseases— brief review.
  42. (2010). Spontaneous generation of mammalian prions.
  43. (2002). Stimulation of PrP(C) retrograde transport toward the endoplasmic reticulum increases accumulation of PrP(Sc) in prion-infected cells.
  44. (1996). Subcellular colocalization of the cellular and scrapie prion proteins in caveolae-like membranous domains.
  45. (1995). The abnormal isoform of the prion protein accumulates in late-endosome-like organelles in scrapie-infected mouse brain.
  46. The comprehensive native interactome of a fully functional tagged prion protein. PLoS One 4,
  47. (2006). The prion protein and lipid rafts.
  48. (2006). The prion protein requires cholesterol for cell surface localization.
  49. (2010). The propagation of prion-like protein inclusions in neurodegenerative diseases.
  50. (1991). The scrapie-associated form of PrP is made from a cell surface precursor that is both protease- and phospholipase-sensitive.
  51. (2002). Transfer of scrapie prion infectivity by cell contact in culture.
  52. (2005). Visualization of prion infection in transgenic mice expressing green fluorescent protein-tagged prion protein.