TGF-β 1, regulation of Alzheimer amyloid precursor protein mRNA expression in a normal human astrocyte cell line: mRNA stabilization

Abstract

The transforming growth factor, TGF-β 1, has been found to be increased in the central nervous system of Alzheimer's disease (AD) patients, elevates amyloid precursor protein (APP) mRNA levels in rat primary astrocytes, and may initiate or promote the deposition of amyloid-β (Aβ) peptide in AD. Excess APP production in AD, which potentially leads to amyloidogenesis, is in part due to over expression of APP mRNA. The production of APP in a normal human cell line in contrast to transformed or animal cells provides a meaningful model to study the regulation of APP gene expression by cytokines that promotes amyloidogenesis. Here, we report that TGF-β 1 treatment of human astrocytes markedly elevated APP mRNA levels, and also increased the half-life of APP message by at least five-fold. Under this condition, as detected by mobility shift and UV cross-linking analysis, a novel 68 kDa RNA–protein complex was formed, involving an 81 nucleotide (nt) fragment within the 3′-untranslated region (UTR), but not the 5′-UTR and coding region of APP mRNA. Insertion of the 3′-UTR onto the chloramphenicol acetyl transferase (CAT) mRNA conferred TGF-β 1 mediated mRNA stability in transfected human astrocytes. On the other hand, the same insert carrying a deletion of the APP mRNA cis-element fragment had no effect on CAT mRNA stability. A model of APP mRNA regulation is presented in which TGF-β 1 induced stabilization of APP message involves the binding activity of a 68 kDa RNA–protein complex within the 3′-UTR, which is likely linked to a reduction in the rate of APP mRNA decay.