Central residues in prion protein PrP^C are crucial for its conversion into the pathogenic isoform

Conformational conversion of the cellular prion protein, PrP^C, into the amyloidogenic isoform, PrP^Sc, is a key pathogenic event in prion diseases. However, the conversion mechanism remains to be elucidated. Here, we generated Tg(PrPΔ91-106)-8545/Prnp^0/0 mice, which overexpress mouse PrP lacking residues 91-106. We showed that none of the mice became sick after intracerebral inoculation with RML, 22L, and FK-1 prion strains nor accumulated PrP^ScΔ91-106 in their brains except for a small amount of PrP^ScΔ91-106 detected in one 22L-inoculated mouse. However, they developed disease around 85 days after inoculation with bovine spongiform encephalopathy (BSE) prions with PrP^ScΔ91-106 in their brains. These results suggest that residues 91-106 are important for PrP^C conversion into PrP^Sc in infection with RML, 22L, and FK-1 prions but not BSE prions. We then narrowed down the residues 91-106 by transducing various PrP deletional mutants into RML- and 22L-infected cells and identified that PrP mutants lacking residues 97-99 failed to convert into PrP^Sc in these cells. Our in vitro conversion assay also showed that RML, 22L, and FK-1 prions did not convert PrPΔ97-99 into PrP^ScΔ97-99, but BSE prions did. We further found that PrP mutants with proline residues at positions 97 to 99 or charged residues at positions 97 and 99 completely or almost completely lost their converting activity into PrP^Sc in RML- and 22L-infected cells. These results suggest that the structurally flexible and noncharged residues 97-99 could be important for PrP^C conversion into PrP^Sc following infection with RML, 22L, and FK-1 prions but not BSE prions.