NR AWEX
AU Fischer,M.; Rogers,J.T.; Rogers,M.
TI Targeting the untranslated regions (UTRs) of the Prnp gene
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions THE-06
PT Konferenz-Poster
AB
Although the underlying mechanism for the conversion of normal cellular prion protein (PrPc) into the pathological isoform (PrPsc) is not fully understood, current strategies for therapy focus on either inhibiting this process or abolishing the formation of PrPsc. PrP0/0 knockout mice lacking the PRNP gene cannot produce the infectious agent, are protected against the disease and show only a subtle phenotype. This indicates suppressing the expression of PrPc may be a reasonable approach to prevent accumulation of PrPsc. We are currently investigating the possibility of specifically downregulating PrPc expression at the translational-level. Using a luciferase-reporter assay with the gene for firefly luciferase under the translational control of the 5'UTR of the human PRNP gene we are monitoring the effect of several compounds on PrP5'UTR-mediated translation in stably transfected SH-SY5Y cells in a 96-well format. Enhanced green fluorescent protein (EGFP) acts as a specific internal control reporting cell viability and transcription levels. A second construct exploiting the 5'UTR of the Alzheimer amyloid precursor protein (APP) gene counterscreens for non-specific hits on PrP5'UTRs. An Iron-responsive Element (IRE) Type II has been found in the APP5'UTR to regulate its translation (Rogers et al., 2002). Compared to APP5'UTR, preliminary data for PrP5'UTR show similar effects on iron treatment in the luciferase assay, suggesting its RNA secondary structure as a possible target for translational regulation (Bandyopadhya et al., 2006). Additional studies indicate a specific regulation of PrP5'UTR-conferred translation. We intend to further confirm and characterise this process by inserting small deletions into the 5'UTR and mapping regions essential for regulation. Compounds identified in a high-throughput screen with FDA-approved drugs, which specifically downC regulate translation mediated by PrP5'UTR, will be tested for their effect on expression of PrP and Sc propagation of PrP in scrapie-infected N2a cells.
Rogers et al. (2002). J Biol Chem 277, 45518-45528 Bandyopadhya et al. (2006). J Biomol Screen. In press
AD M. Fischer, M. Rogers: Prion Research Group, School of Biology and Environmental Science, University College Dublin, Dublin, Ireland; J.T. Rogers: Genetics and Aging Research Unit, Psychiatry Department, Massachusetts General Hospital, Cambridge MA, USA. E-mail: mark.rogers@ucd.ie
SP englisch
PO Italien