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J Biol Chem, Vol. 274, Issue 52, 37177-37185, December 24, 1999

Transcriptional Induction of Stromelysin-3 in Mesodermal Cells Is Mediated by an Upstream CCAAT/Enhancer-binding Protein Element Associated with a DNase I-hypersensitive Site*

Daochun LuoDagger , Eric GuérinDagger , Marie-Gabrielle Ludwig, Isabelle Stoll, Paul Basset, and Patrick Anglard§

From the Institut de Génétique et de Biologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, INSERM, Université Louis Pasteur, Communauté Urbaine de Strasbourg, 67404 Illkirch cedex, France

Stromelysin-3 (ST3) is a matrix metalloproteinase whose synthesis is markedly increased in stromal fibroblasts of most invasive human carcinomas. In the present study, we have investigated the molecular mechanisms by which high levels of ST3 expression can be induced. In contrast to the early and transient induction of interstitial collagenase by 12-O-tetradecanoylphorbol-13-acetate (TPA), the fibroblastic induction of ST3 was found to be delayed and to require protein neosynthesis. We demonstrated that this induction is transcriptional and does not result from changes in RNA stability. By looking next to promoter regions accessible to DNase I upon gene induction, we have identified two distal elements and have characterized their role in the transcriptional regulation of ST3. The first one is a TPA-responsive element that controls the base-line ST3 promoter activity but is not required for its activation. We demonstrate that ST3 gene induction is actually mediated by the second element, a C/EBP-binding site, by showing: (i) that this element becomes accessible in cells induced to express ST3, (ii) that endogenous C/EBPbeta binds to the ST3 promoter, and (iii) that this binding leads to ST3 transcriptional activation. Our study provides new insights into the regulation of ST3 and suggests an additional role for C/EBP transcription factors in tissue remodeling processes associated with this MMP.

* This work was supported by funds from INSERM, the Center National de la Recherche Scientifique, the Hôpital Universitaire de Strasbourg, and the Bristol-Myers Squibb Pharmaceutical Research Institute, by BIOMED 2 Grant BMH4CT96-0017, BIOTECH 2 Grant ERBBIO4CT96-0464, and Association pour la Recherche sur le Cancer Grant 9185, and by funds from the Ligue Nationale Française contre le Cancer, the Comité du Haut-Rhin, and the Fondation pour la Recherche Médicale Française.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.

Dagger These authors contributed equally to this work.

§ To whom correspondence should be addressed: Inst. de Génétique et de Biologie Moléculaire et Cellulaire, BP 163, 67404 Illkirch cedex, France. Tel.: 33-388-65-3421; Fax: 33-388-65-3201; E-mail: anglard@titus.u-strasbg.fr.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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