Kinetics of expression of prion protein in uninfected and scrapie‐infected N₂a mouse neuroblastoma cells

Abstract

The scrapie prion protein, PrPSc, is formed from its isoform, the cellular PrPc. There is evidence available indicating that PrPSc is a necessary component of the infectious prion particle to cause a series of transmissible spongiform encephalopathies. We have used immunocytochemistry and RNA blotting techniques to investigate if infection with prions results in an increased PrP gene expression. For the experiments we used N2a cells which had been infected with prions (ScN2a cells). We demonstrated by confocal laser scanning microscopy that PrP-protein was present in the nucleus (predominantly in the nucleoli) of ScN2a cells. Analysis of the PrP-mRNA levels both in N2a- and in ScN2a cells using cDNA encoding PrPc revealed no marked alteration of the mRNA steady state level between the two cell strains. Likewise, in run-off experiments no changes in either PrP-specific transcription or in general transcriptional activity were found. The half-life of PrP-mRNA was found to be identical in both cell strains (7h). Taken together, these results show that PrPSc and/or PrPc is present in the nucleus (nucleoli) of ScN2a cells but does not display an effect on the expression of the PrP gene.

Stimulation of PrPC Retrograde Transport toward the Endoplasmic Reticulum Increases Accumulation of PrPSc in Prion-infected Cells

“…These results were independent of the N2a cell line, because similar results were observed with Chinese hamster ovary cells overexpressing MoPrP (data not shown). Interestingly, we can also detect the presence of PrP in the nuclei of the cells, presumably in the nucleoli, as already described (18,19).…”

Specific inhibition of pathological prion protein accumulation by small interfering RNAs

“…In neuroblastoma cells, PrP-mRNAs have a half-life of approximately 7 hours (Pfeifer et al, 1993) and PrP-sen is synthesized and degraded relatively rapidly (tg ~5 hours). Whereas, PrP-res aggregates which derived from PrP-sen, is (0) or in the presence of increasing concentrations of Prnp gene-specific siRNA or of scrambled siRNA duplexes.…”

Differential Display Detects Host Nucleic Acid Motifs Altered in Scrapie-Infected Brain

“…PrP sc is translocated into nuclei 73 where confocal laserscanning microscopy on infected neuroblastoma cells has revealed that PrP and/or PrP sc preferentially reside within nucleoli. 74 Biochemical association of ros sequences with nucleoli may therefore favor PrP interactions.…”