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Originally published In Press as doi:10.1074/jbc.M105057200 on November 15, 2001

J. Biol. Chem., Vol. 277, Issue 7, 5514-5523, February 15, 2002
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Modulation of Apolipoprotein D and Apolipoprotein E mRNA Expression by Growth Arrest and Identification of Key Elements in the Promoter*

Sonia Do CarmoDagger , Diane SéguinDagger , Ross Milne§, and Eric RassartDagger

From the Dagger  Laboratoire de biologie moléculaire, Département des Sciences Biologiques, Université du Québec à Montréal, Montréal H3C 3P8, Québec and the § Lipoprotein and Atherosclerosis Research Group, University of Ottawa Heart Institute, 40 Ruskin St., Ottawa, Ontario K1Y 4W79, Canada

Apolipoprotein D (apoD) and apolipoprotein E (apoE) are co-expressed in many tissues, and, in certain neuropathological situations, their expression appears to be under coordinate regulation. We have previously shown that apoD gene expression in cultured human fibroblasts is up-regulated when the cells undergo growth arrest. Here, we demonstrate that, starting around day 2 of growth arrest, both apoD and apoE mRNA levels increase between 1.5- and 27-fold in other cell types, including mouse primary fibroblasts and fibroblast-like and human astrocytoma cell lines. To understand the regulatory mechanisms of apoD expression, we have used apoD promoter-luciferase reporter constructs to compare gene expression in growing cells and in cells that have undergone growth arrest. Analysis of gene expression in cells transfected with constructs with deletions and mutations in the apoD promoter and constructs with artificial promoters demonstrated that the region between nucleotides -174 and -4 is fully responsible for the basal gene expression, whereas the region from -558 to -179 is implicated in the induction of apoD expression following growth arrest. Within this region, an alternating purine-pyrimidine stretch and a pair of serum-responsive elements (SRE) were found to be major determinants of growth arrest-induced apoD gene expression. Evidence is also presented that SREs in the apoE promoter may contribute to the up-regulation of apoE gene expression following growth arrest.

* This work was supported by the Canadian Institute for Health Research (grants MT-9880 and MT-15677) and by the Heart and Stroke Foundation of Canada.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Laboratoire de Biologie Moléculaire, Département des Sciences Biologiques, CP 8888 Succ Centre-ville, Université du Québec à Montréal, Montréal H3C 3P8, Québec. Tel.: 514-987-3000 (Ext. 3953); Fax: 514-987-4647; E-mail: Rassart.Eric@UQAM.CA.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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