Role of cytokines in the gene expression of amyloid β–protein precursor: Identification of a 5'-UTR-Binding nuclear factor and its implications in Alzheimer's disease
Affiliations: [a] Department of Psychiatry, Institute of Psychiatric Research, 791 Union Drive, Indiana University School of Medicine, Indianapolis, IN--46202, USA | [b] UMR CNRS 6551, Université de Caen, Cyceron, Bd, Cedex France | [c] Laboratory of Neurosciences, National Institute on Aging, Baltimore, MD 21224, USA | [d] Genetics and Aging Unit, Harvard Medical School, Boston, MA 02129, USA
Correspondence:
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Corresponding author: Debomoy K. Lahiri, Ph.D., Indiana University School of Medicine, Institute of Psychiatric Research, Rm. PR-313, 791 Union Drive, Indianapolis, IN 46202-4887, USA. Tel.: +1 317 274 2706; Fax: +1 317 274 1365; E-mail: dlahiri@iupui.edu.
Abstract: One of the major neuropathological characteristics of Alzheimer's disease (AD) is the brain depositions of senile plaques that are mainly composed of toxic amyloid β–peptide (Aβ), which is generated from a family of Aβ containing precursor proteins (AβPP; 695-770 amino acids). The role of cytokines and growth factors has been implicated in the pathogenesis of AD. Our goal is to determine the mode of action of cytokines on the regulation of AβPP gene expression. Here we studied the effect of different cytokines on the activity of 5'-untranslated region (5'-UTR) of AβPP mRNA in human astrocytic cells (U-373). We compared AβPP-5'-UTR activity in the presence of interleukin-1 (IL-1α and IL-1β), transforming growth factor (TGF-β1) and tumor necrosis factor TNF-α1. The astrocytic cells, which were treated separately with these agents, were transfected with either the vector (pSV2CAT) or pSV2UTR-CAT construct containing 90 bp of AβPP 5'-UTR +54 to 144 bp). This region was cloned upstream of a reporter chloramphenicol acetyl transferase gene (CAT). Our results indicate that the treatment of pSV2UTR-CAT-transfected cells with either IL-1α, IL-1β, TGF-β1 or TNF-α1 stimulated reporter gene activity in a factor-specific manner. This was consistent with their effects on elevating AβPP protein levels. Transfection of the same cells with the pSV2CAT vector lacking 5'-UTR resulted in a reduced reporter gene activity with all treatments studied. DNA-gel shift experiments indicate that the 54/144 region binds to a nuclear protein(s) in a cell type specific manner. These results suggest that 5'-UTR of the AβPP gene can respond to the stimulation of different cytokines, which likely regulate AβPP transcription and translation via regulatory elements present in the AβPP promoter and in 5'-UTR, respectively. The characterization of AβPP regulatory elements, including the 5'-UTR, will accelerate the development of novel agents against new targets for AD.
