Functional characterization of the human prion protein promoter in neuronal and endothelial cells

Citation
H. Funke-kaiser et al., Functional characterization of the human prion protein promoter in neuronal and endothelial cells, J MOL MED-J, 79(9), 2001, pp. 529-535
Citations number
53
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Research/Laboratory Medicine & Medical Tecnology","Medical Research General Topics
Journal title
JOURNAL OF MOLECULAR MEDICINE-JMM
ISSN journal
0946-2716 → ACNP
Volume
79
Issue
9
Year of publication
2001
Pages
529 - 535
Database
ISI
SICI code
0946-2716(200109)79:9<529:FCOTHP>2.0.ZU;2-J
Abstract
Human prion diseases such as Creutzfeld-Jakob disease and kuru are of major medical and biological importance because of their fatal course, epidemic potential, and unique pathophysiology. Endogenous expression of the normal cellular prion protein (PrPC) is necessary for infection and prion replicat ion. However, knowledge of human PrPC gene regulation is rudimentary. We th erefore cloned1543 bp of the 5' untranslated and promoter region of the PrP gene. Using transient transfection assays, the full-length promoter and se rial deletion mutants subcloned in a luciferase reporter vector were analyz ed in neuronal (KELLY) and endothelial (EA.hy926) cell lines, which both ex press PrPC as shown by RT/PCR. Analysis of promoter constructs in KELLY cel ls indicated two activating regions at -131/-284 and -1303/-1543, relative to the 3'-terminal end of exon 1, and also two repressing elements at -254/ -567 and -567/-909 in neuronal cells. In EA.hy926 cells, activating element s were identified at -131/-284 and -284/-567, and one repressing region was localized at -567/-909. In addition, transcriptional start sites were dete rmined by 5'-RACE reaction and RNase protection assay, revealing one major transcriptional start site located at -47 (in KELLY cells), -53 (in human t halamus) and at about -55 (in EA.hy926 cells).