Amino acid sequence and prion strain specific effects on the in vitro and in vivo convertibility of ovine/murine and bovine/murine prion protein chimeras

Abstract

Prion diseases are characterised by the conversion of a cellular prion protein (PrP C) by its misfolded, hence pathogenic, isoform (PrP Sc). The efficiency of this transition depends on the molecular similarities between both interaction partners and on the intrinsic convertibility of PrP C. Transgenic mice expressing chimeric murine/ovine PrP C (Tgmushp mice) are susceptible to BSE and/or scrapie prions of bovine or ovine origin while transgenic mice expressing similar murine/bovine PrP C chimera (Tgmubo mice) are essentially resistant. We have studied this phenomenon by cell-free conversion on procaryotically expressed chimeric PrP C. Mouse passaged scrapie or BSE PrP Sc was used as a seed and the conversion reaction was carried out under semi-native conditions. The results obtained in this assay were similar to those of our in vivo experiments. Since mubo- and mushp-PrP C differ only at four amino acid positions (S96G, N142S, Y154H and Q185E), single or double point mutations of mushp-PrP C were examined in the cell-free conversion assay. While the scrapie Me7 prion induced conversion was largely reduced by the N142S and Q185E but not by the S96G and Y154H mutation, the BSE induced conversion was retained in all mutants. Newly formed PrP res exhibited strain specific characteristics, such as the localisation of the proteinase K cleavage site, even in the chimeric PrP C mutants. We therefore postulate that the efficiency of the conversion of chimeric PrP C depends on the amino acid sequence as well as on prion strain specific effects.