NR AXTB
AU Mossavitzky,T.; Rouvinski,A.; Metzer,E.; Gahali-Sass,I.; Luhr,K.; Kristensson,K.; Taraboulos,A.
TI Transcriptionally-active Drugs Increase the Scrapie Prion Protein under the control of a CMV promoter: Fundamental and Practical Implications
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Protein Misfolding P01.40
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB Prion-infected cell lines typically produce modest levels of PrPsc. In mouse neuroblastoma ScN2a cells, about 5% of PrPc is transformed into PrPsc. In this study we have used the histone deacetylase inhibitor trichostatin A (TSA) as well as other transcriptionally active drugs to increase the exogenous expression of epitopicallylabeled PrPc-MHM2 controlled by the early-late CMV promoter. In N2a-MHM2 cells, 3µM TSA (16h) increased both PrP mRNA and PrPc expression by about 10 times. Significantly, in ScN2a-MHM2, the transgenic PrPsc-MHM2 also increased by about 10x, showing (i) that PrPc level is a limiting factor for PrPsc production, and (ii) that the metabolism leading to PrPsc formation is not hampered by TSA. In the GT1-MHM2 system, the effect of TSA was more modest, resulting in a 3-fold increase of PrPc and PrPsc. In both cell lines, the PrP increase was concentration-dependent and peaked at micromolar TSA concentrations. Longer TSA treatments or higher concentrations were toxic. Of note, TSA also somewhat increased PrPsc (but not PrPc) in non-transgenic ScGT1, pointing to a modulation of PrPsc metabolism by transcriptionally active drugs. Other drugs and their synergism were also investigated. Increasing PrPsc production by manipulating the CMV promoter may help to study the metabolism of prions and will improve the sensitivity of cell culture-based prion bioassays (supported by awards US-ARMRC DAMD17-03-102288 and EC QLK2-CT-2002-81628 ).
AD T. Mossavitzky, A. Rouvinski, E. Metzer, I. Gahali-Sass, A. Taraboulos, Hebrew University-Hadassah Medical School and the Israel Prion Center, Department of Molecular Biology, Israel; K. Luhr, K. Kristensson, Karolinska Institutet, Department of Neuroscience, Sweden
SP englisch
PO Schottland