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Originally published In Press as doi:10.1074/jbc.M006215200 on December 5, 2000

J. Biol. Chem., Vol. 276, Issue 10, 7101-7107, March 9, 2001
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Cbfa1 Contributes to the Osteoblast-specific Expression of type I collagen Genes*

Britt Kern, Jianhe Shen, Michael Starbuck, and Gerard KarsentyDagger

From the Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030

Type I collagen is composed of two chains, alpha 1(I) and alpha 2(I), encoded by two distinct genes, the alpha 1(I) and alpha 2(I) collagen genes, that are highly expressed in osteoblasts. In most physiological situations, alpha 1(I) and alpha 2(I) collagen expression is coregulated, suggesting that identical transcription factors control their expression. Here, we studied the role of Cbfa1, an osteoblast-specific transcription factor, in the control of alpha 1(I) and alpha 2(I) collagen expression in osteoblasts. A consensus Cbfa1-binding site, termed OSE2, is present at the same location in the alpha 1(I) collagen promoter at approximately -1347 base pairs (bp) of the rat, mouse, and human genes. Cbfa1 can bind to this site, as demonstrated by electrophoretic mobility shift assay (EMSA) and supershift experiments using an anti-Cbfa1 antibody. Mutagenesis of the alpha 1(I) collagen OSE2 at -1347 bp reduced the activity of a alpha 1(I) collagen promoter fragment 2- to 3-fold. Moreover, multimers of this OSE2 at -1347bp confer osteoblast-specific activity to a minimum alpha 1(I) collagen promoter fragment in DNA transfection experiments as well as in transgenic mice. An additional Cbfa1-binding element is present in the alpha 1(I) collagen promoter of mouse, rat, and human at approximately position -372. This site binds Cbfa1 only weakly and does not act as a cis-acting activator of transcription when tested in DNA transfection experiments. Similar to alpha 1(I) collagen, the mouse alpha 2(I) collagen gene contains multiple OSE2 sites, of which one is conserved across multiple species. In EMSA, Cbfa1 binds to this site and multimers of this alpha 2(I) OSE2 element confer osteoblast-specific activity to the minimum alpha 1(I) collagen promoter in DNA transfection experiments. Thus, our results suggest that Cbfa1 is one of the positive regulators of the osteoblast-specific expression of both type I collagen genes.

* This work was supported by National Institutes of Health Grants R01AR45548 and R01DE11290, March of Dimes Grant F-198-0082, and a grant from Eli Lilly and Co. (to G. K.).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 To whom correspondence should be addressed: Dept. of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030. Tel.: 713-798-5489; Fax: 713-798-1465; E-mail karsenty@bcm.tmc.edu.

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


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