Comparative sequence analysis, in vitro expression and biosynthesis of mouse PrP.
A number of studies have indicated that an endogenous brain protein, PrP, is associated with transmissible agents causing spongiform encephalopathies such as scrapie, kuru, and Creutzfeldt-Jakob disease. It has been proposed that PrP derived from scrapie brain is the scrapie agent itself. To test directly whether the PrP mRNA in scrapie brain tissue can encode the scrapie agent, we expressed PrP cDNA cloned from scrapie-infected mouse brain in vitro. The expressed PrP did not transmit scrapie to susceptible mice. Thus either PrP is not the scrapie agent, or the expressed PrP requires additional modification to be infectious. The normal function of PrP is unknown, however, comparison of the amino acid sequences of PrP from mouse, hamster, and human revealed that many structural features of potential functional significance have been conserved during evolution. To learn about normal PrP and whether it is altered by scrapie infection in vitro, we have performed studies of PrP biosynthesis in normal and scrapie-infected mouse neuroblastoma tissue culture cells. The major PrP species were glycoproteins anchored at the cell surface by covalent linkage to phosphatidylinositol. No scrapie-associated modifications of PrP biosynthesis were observed, and, none of the metabolically labeled PrP observed in either scrapie-infected normal cells was resistant to proteinase K.
Caughey
B
B
Laboratory of Persistent Viral Diseases, National Institute of Allergy and Infectious Diseases, Hamilton, MT 59840.
Race
R
R
Chesebro
B
B
D000595:Amino Acid Sequence; D000818:Animals; D002478:Cells, Cultured; D002999:Clone Cells; D006224:Cricetinae; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D018622:PrP 27-30 Protein; D011328:Prions / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger; D014764:Viral Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*
D011328:Prions; D012333:RNA, Messenger; D014764:Viral Proteins; D018622:PrP 27-30 Protein