Cloning and expression of the rat BACE1 promoter

Christine Lange-Dohna, Ulrike Zeitschel, Frank Gaunitz, J. Regino Perez-Polo, Volker Bigl, Steffen Roßner

    Research output: Contribution to journalArticle

    45 Citations (Scopus)

    Abstract

    The pathogenic processing of the amyloid precursor protein (APP) into β-amyloid peptides, which give rise to β-amyloid plaques in the brains of Alzheimer's disease patients, requires the enzymatic activity of the beta-site APP-cleaving enzyme 1 (BACE1). We report the cloning and sequence of a 1.5-kb DNA fragment upstream of the coding sequence of the rat BACE1 gene and the construction of a BACE1 promoter/luciferase reporter construct. The basal activity of this promoter construct was highest in neuronal cell lines such as BE(2)-C and PC12 and in the pancreatic cell line AR42J, somewhat lower in rat primary neurons, and astrocytic and microglial cultures, very low in hepatocytes, and almost absent in fibroblasts and in the monocyte-macrophage cell line RAW264.7. The first 600 bp of this promoter are highly conserved among rat, mouse, and human, suggesting that this region contains regulatory elements that modulate BACE1 transcription. Indeed, this fragment contains several putative transcription factor binding sites such as MZF1, Sp1, four GATA-1 sites, and one YY1 site. Directed mutagenesis of GATA-1 elements led to altered luciferase expression, indicating that these sites are involved in the regulation of BACE1 transcription. Additionally, the analysis of promoter activities of deletion mutants suggests the presence of activators of BACE1 transcription between bases -514 to -753 and of suppressor elements between bases -754 and -1541. The BACE1 promoter sequence data and the constructs described here will be useful to identify factors that influence the expression of BACE1 in experimental paradigms in vitro.

    Original languageEnglish (US)
    Pages (from-to)73-80
    Number of pages8
    JournalJournal of Neuroscience Research
    Volume73
    Issue number1
    DOIs
    StatePublished - Jul 1 2003

    Fingerprint

    Organism Cloning
    Enzymes
    Amyloid beta-Protein Precursor
    Luciferases
    Cell Line
    Nucleic Acid Regulatory Sequences
    Amyloid Plaques
    Brain Diseases
    rat Bace protein
    Amyloid
    Mutagenesis
    Monocytes
    Hepatocytes
    Alzheimer Disease
    Transcription Factors
    Fibroblasts
    Macrophages
    Binding Sites
    Neurons
    Peptides

    Keywords

    • β-amyloid
    • β-secretase
    • Alzheimer's disease
    • Amyloid precursor protein
    • Directed mutagenesis
    • Luciferase assay
    • Promoter construct

    ASJC Scopus subject areas

    • Neuroscience(all)

    Cite this

    Lange-Dohna, C., Zeitschel, U., Gaunitz, F., Perez-Polo, J. R., Bigl, V., & Roßner, S. (2003). Cloning and expression of the rat BACE1 promoter. Journal of Neuroscience Research, 73(1), 73-80. https://doi.org/10.1002/jnr.10639

    Cloning and expression of the rat BACE1 promoter. / Lange-Dohna, Christine; Zeitschel, Ulrike; Gaunitz, Frank; Perez-Polo, J. Regino; Bigl, Volker; Roßner, Steffen.

    In: Journal of Neuroscience Research, Vol. 73, No. 1, 01.07.2003, p. 73-80.

    Research output: Contribution to journalArticle

    Lange-Dohna, C, Zeitschel, U, Gaunitz, F, Perez-Polo, JR, Bigl, V & Roßner, S 2003, 'Cloning and expression of the rat BACE1 promoter', Journal of Neuroscience Research, vol. 73, no. 1, pp. 73-80. https://doi.org/10.1002/jnr.10639
    Lange-Dohna C, Zeitschel U, Gaunitz F, Perez-Polo JR, Bigl V, Roßner S. Cloning and expression of the rat BACE1 promoter. Journal of Neuroscience Research. 2003 Jul 1;73(1):73-80. https://doi.org/10.1002/jnr.10639
    Lange-Dohna, Christine ; Zeitschel, Ulrike ; Gaunitz, Frank ; Perez-Polo, J. Regino ; Bigl, Volker ; Roßner, Steffen. / Cloning and expression of the rat BACE1 promoter. In: Journal of Neuroscience Research. 2003 ; Vol. 73, No. 1. pp. 73-80.
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