title: Glycoform-independent prion conversion by highly efficient, cell-based, protein misfolding cyclic amplification.
abstract: Prions are formed of misfolded assemblies (PrP(Sc)) of the variably N-glycosylated cellular prion protein (PrP(C)). In infected species, prions replicate by seeding the conversion and polymerization of host PrP(C). Distinct prion strains can be recognized, exhibiting defined PrP(Sc) biochemical properties such as the glycotype and specific biological traits. While strain information is encoded within the conformation of PrP(Sc) assemblies, the storage of the structural information and the molecular requirements for self-perpetuation remain uncertain. Here, we investigated the specific role of PrP(C) glycosylation status. First, we developed an efficient protein misfolding cyclic amplification method using cells expressing the PrP(C) species of interest as substrate. Applying the technique to PrP(C) glycosylation mutants expressing cells revealed that neither PrP(C) nor PrP(Sc) glycoform stoichiometry was instrumental to PrP(Sc) formation and strainness perpetuation. Our study supports the view that strain properties, including PrP(Sc) glycotype are enciphered within PrP(Sc) structural backbone, not in the attached glycans.
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Moudjou
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Chapuis
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Mériem
authors.initials: M
authors.lastname: Mekrouti
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Fabienne
authors.initials: F
authors.lastname: Reine
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Herzog
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Pierre
authors.initials: P
authors.lastname: Sibille
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Hubert
authors.initials: H
authors.lastname: Laude
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.|IHAP, INRA, Ecole Nationale Vétérinaire de Toulouse, 31000, Toulouse, France.
authors.forename: Didier
authors.initials: D
authors.lastname: Vilette
authors.affiliation: IHAP, INRA, Ecole Nationale Vétérinaire de Toulouse, 31000, Toulouse, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Andréoletti
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Human
authors.initials: H
authors.lastname: Rezaei
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Dron
authors.affiliation: VIM, INRA, Université Paris-Saclay, 78350, Jouy-en-Josas, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Béringue
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods*; D001921:Brain / Q000378:metabolism; D002457:Cell Extracts; D002460:Cell Line; D002478:Cells, Cultured; D004586:Electrophoresis; D055786:Gene Knockout Techniques; D006031:Glycosylation; D006801:Humans; D008822:Mice, Transgenic; D008863:Microspheres; D008904:Miniaturization; D050505:Mutant Proteins / Q000378:metabolism; D011328:Prions / Q000378:metabolism*; D017510:Protein Folding; D013997:Time Factors
chemical_list: D002457:Cell Extracts; D050505:Mutant Proteins; D011328:Prions