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Originally published In Press as doi:10.1074/jbc.M104185200 on July 25, 2001

J. Biol. Chem., Vol. 276, Issue 40, 37011-37019, October 5, 2001
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delta EF1 Binds to a Far Upstream Sequence of the Mouse Pro-alpha 1(I) Collagen Gene and Represses Its Expression in Osteoblasts*

Catherine TerrazDagger §, Dave Toman, Madeleine DelaucheDagger , Pierre RoncoDagger , and Jerome RossertDagger ||

From the Dagger  INSERM U489 and Université Paris VI, Paris, France and  Cohesion Technologies, Palo Alto, California 94303

The transcription of type I collagen genes is tightly regulated, but few cis-acting elements have been identified that can modulate the levels of expression of these genes. Generation of transgenic mice harboring various segments of the mouse pro-alpha 1(I) collagen promoter led us to suspect that a repressor element was located between -10.5 and -17 kilobase pairs. Stable and transient transfection experiments in ROS17/2.8 osteoblastic cells confirmed the existence of such a repressor element at about -14 kilobase pairs and showed that it consisted in an almost perfect three-time repeat of a 41-base pair sequence. This element, which we named COIN-1, contains three E2-boxes, and a point mutation in at least two of them completely abolished its repressor effect. In gel shift assays, COIN-1 bound a DNA-binding protein named delta EF1/ZEB-1, and mutations that abolished the repressor effect of COIN-1 also suppressed the binding of delta EF1. We also showed that the repressor effect of COIN-1 was not mediated by chromatin compaction. Furthermore, overexpression of delta EF1 in ROS17/2.8 osteoblastic cells enhanced the inhibitory effect of COIN-1 in a dose-dependent manner and repressed the expression of the pro-alpha 1(I) collagen gene. Thus, delta EF1 appears to repress the expression of the mouse pro-alpha 1(I) collagen gene, through its binding to COIN-1.

* This work was supported by grants from the Association pour la Recherche sur le Cancer (to J. R.) and from the University of Paris (to J. R.).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.

§ Recipient of a fellowship from the Ministère de l'Education, de la recherche, et de la Technologie.

|| To whom correspondence should be addressed: INSERM U489, Hôpital TENON, 4 rue de la Chine, 75020 Paris, France. Tel.: 33-1-56-01-69-93; Fax: 33-1-56-01-69-99; E-mail: jerome.rossert@tnn.ap-hop-paris.fr.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.


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