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Mechanism of promoter activity of the β‐amyloid precursor protein gene in different cell lines: identification of a specific 30 bp fragment in the proximal promoter region
Author(s) -
Ge YuanWen,
Ghosh Chandramallika,
Song Weihong,
Maloney Bryan,
Lahiri Debomoy K.
Publication year - 2004
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.2004.02608.x
Subject(s) - microbiology and biotechnology , transfection , biology , gene , gene expression , amyloid precursor protein , cell culture , reporter gene , electrophoretic mobility shift assay , promoter , nuclear protein , regulation of gene expression , transcription factor , alzheimer's disease , genetics , pathology , medicine , disease
The amyloid β‐protein (Aβ) deposited in brains of Alzheimer's disease (AD) patients is proteolytically derived from a large Aβ precursor protein (APP). APP gene expression patterns in the AD brain region indicate that abnormalities of gene regulation may be important in AD pathology. To understand the contribution of different cell types to APP gene expression, we studied it at four levels: promoter activity (by reporter gene assay of transfected cells), DNA–nuclear protein interaction (by electrophoretic mobility shift assay), RNA message and protein (by northern and western blotting, respectively). APP mRNA and protein expression levels were greater in neuroblastoma and PC12 cells than in glial or cervix epithelial cells. Relative activity among 12 different promoter regions and within single regions varied according to cell type/cell line. An upstream regulatory region containing a GATA‐1 site is necessary for activity in PC12 and glial cells but not in neuroblastoma cells. DNA–protein interactions were examined in three distal and one proximal promoter elements in nuclear extracts belonging to neuronal and non‐neuronal cells. The proximal promoter region is important for cell line‐specific APP gene expression. Characterization of the APP regulatory region's interaction with cell type‐specific nuclear factor(s) is important to understand tissue‐specific expression of APP seen in AD subjects.