title: Rapid cell-surface prion protein conversion revealed using a novel cell system.
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 (PrP(C)). Here we develop a unique cell system in which epitope-tagged PrP(C) is expressed in a PrP knockdown (KD) neuroblastoma cell line. The tagged PrP(C), when expressed in our PrP-KD cells, supports prion replication with the production of bona fide epitope-tagged infectious misfolded PrP (PrP(Sc)). Using this epitope-tagged PrP(Sc), 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.
authors.affiliation: Department of Neurodegenerative Disease, Institute of Neurology, University College London, Queen Square, London WC1N 3BG, UK.
authors.forename: R
authors.initials: R
authors.lastname: Goold
authors.forename: S
authors.initials: S
authors.lastname: Rabbanian
authors.forename: L
authors.initials: L
authors.lastname: Sutton
authors.forename: R
authors.initials: R
authors.lastname: Andre
authors.forename: P
authors.initials: P
authors.lastname: Arora
authors.forename: J
authors.initials: J
authors.lastname: Moonga
authors.forename: A R
authors.initials: AR
authors.lastname: Clarke
authors.forename: G
authors.initials: G
authors.lastname: Schiavo
authors.forename: P
authors.initials: P
authors.lastname: Jat
authors.forename: J
authors.initials: J
authors.lastname: Collinge
authors.forename: S J
authors.initials: SJ
authors.lastname: Tabrizi
mesh_terms: D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005455:Fluorescent Antibody Technique; D055785:Gene Knockdown Techniques; D008565:Membrane Proteins / Q000737:chemistry*; D051379:Mice; D017096:Prion Diseases / Q000378:metabolism*; D011328:Prions / Q000737:chemistry* / Q000235:genetics; D017510:Protein Folding; D021381:Protein Transport; D034622:RNA Interference; D013997:Time Factors
chemical_list: D008565:Membrane Proteins; D011328:Prions