Prion propagation is dependent on key amino acids in Charge cluster 2 within the prion protein

Autor: Savroop Bhamra, Parineeta Arora, Szymon W. Manka, Christian Schmidt, Craig Brown, Melissa L. D. Rayner, Peter-Christian Klöhn, Anthony R. Clarke, John Collinge, Parmjit S. Jat
Rok vydání: 2022
Předmět:
DOI: 10.1101/2022.08.08.503133
Popis: Prions consist of assemblies of aberrantly folded cellular prion protein (PrPC) upon template-assisted conversion and propagation of disease-associated PrP. To dissect the N-terminal residues critical for efficient prion propagation, we generated a library of point, double, or triple alanine replacements within residues 23-111 of PrP, stably expressed them in cells silenced for endogenous mouse PrPC and challenged the reconstituted cells with four mouse prion strains. Amino acids (aa) 105-111 of Charge Cluster 2 (CC2), which is disordered in PrPC, were required for propagation of all four prion strains; other residues had no effect or exhibited strain-specific effects. Replacements in CC2, including aa105-111, dominantly inhibited prion propagation in the presence of endogenous wild type PrPC whilst other changes were not inhibitory. Single alanine replacements within aa105-111 identified leucine 108 and valine 111 or the cluster of lysine 105, threonine 106 and asparagine 107 as critical for prion propagation. These residues mediate specific ordering of CC2 in the prion fibrils from Rocky Mountain Laboratory (RML) and ME7 mouse prion strains.
Databáze: OpenAIRE