In vitro expression and biosynthesis of prion protein.
In addition to whatever function PrP may have normally, its involvement in scrapie-like neurodegenerative diseases has become clearer in recent years. In vitro studies have made important contributions to the understanding of normal PrP biosynthesis and turnover and how they can be influenced by scrapie infection. Cell-free transcription and translation experiments have indicated that PrP gene translation products are capable of assuming two different topologies, one spanning microsomal membranes and the other completely translocated into the microsomal lumen (Hay et al. 1987a, b). A novel stop transfer signal in the polypeptide is critical to the formation of the transmembrane topology (Yost et al. 1990). Expression of recombinant PrP genes has been accomplished in mouse (Caughey et al. 1988b), monkey (Scott et al. 1988), frog (Hay et al. 1987a), and insect (Scott et al. 1988) tissue culture cells. PrP products encoded by PrP cDNAs cloned from scrapie-infected brain tissues are not infectious and do not have the protease-resistance characteristic of the scrapie-associated form of PrP isolated from diseased tissue (Caughey et al. 1988b; Scott et al. 1988). Studies of PrP encoded by the endogenous gene of mouse neuroblastoma cells have identified the precursors (Caughey et al. 1989) and products (Race et al. 1988; Caughey et al. 1989) of normal PrP biosynthesis and shown that most of the PrP of normal cells is linked to the cell surface by phosphatidylinositol (Stahl et al. 1987; Caughey et al. 1989, 1990; Borchelt et al. 1990). In scrapie-infected clones, and additional pool of PrP is present which, unlike the normal PrP, aggregates (B. Caughey, unpublished observations) and is partially protease resistant (Butler et al. 1988; Caughey et al. 1990; Borchelt et al. 1990; Stahl et al. 1990). This scrapie-associated pool of PrP differs from the normal PrP in that it is primarily intracellular (Caughey et al. 1990; Borchelt et al. 1990; Taraboulos et al. 1990) and resistant to removal from cells by phospholipase or protease (Caughey et al. 1990; Borchelt et al. 1990; Stahl et al. 1990) treatments. Kinetic studies have shown that while PrP-sen is synthesized and degraded relatively rapidly (Caughey et al. Borchelt et al. 1990), PrP-res is synthesized slowly and has a very long half-life (Borchelt et al. 1990). Further studies with the scrapie-infected mouse neuroblastoma cells should lead toward the elucidation of the molecular details of the scrapie-associated modification of PrP and whether the modification is directly related to scrapie agent replication.(ABSTRACT TRUNCATED AT 400 WORDS)
Caughey
B
B
Laboratory of Persistent Viral Diseases, National Institute of Allergy and Infectious Diseases, Hamilton, Mt 59840.
D000818:Animals; D002460:Cell Line; D002474:Cell-Free System; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D009036:Moths / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000235:genetics; D009447:Neuroblastoma / Q000473:pathology; D018620:PrPSc Proteins; D011328:Prions / Q000096:biosynthesis* / Q000235:genetics; D011487:Protein Conformation; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis; D012608:Scrapie / Q000378:metabolism / Q000473:pathology; D012756:Sheep; D013045:Species Specificity; D013347:Subcellular Fractions / Q000378:metabolism; D014908:Triticum / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000378:metabolism; D014714:Vertebrates / Q000235:genetics / Q000378:metabolism
D006023:Glycoproteins; D008562:Membrane Glycoproteins; D009419:Nerve Tissue Proteins; D018620:PrPSc Proteins; D011328:Prions; D011498:Protein Precursors; D011994:Recombinant Proteins