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Papers In Press, published online ahead of print December 5, 2000
J. Biol. Chem, 10.1074/jbc.M006215200
Submitted on July 13, 2000
Revised on December 5, 2000
Accepted on December 4, 2000
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030
Corresponding Author: karsenty{at}bcm.tmc.edu
Type I collagen is composed of two chains, 
1(I) and 2(I), encoded by two distinct genes, the 1(I) and 2(I) collagen genes, that are highly expressed in osteoblasts. In most physiological situations, 1(I) and 2(I) collagen expression are co-regulated, suggesting that identical transcription factors control their expression. Here, we studied the role of Cbfa1, an osteoblast-specific transcription factor, in the control of 1(I) and 2(I) collagen expression in osteoblasts. A consensus Cbfa1-binding site, termed OSE2, is present at the same location in the 1(I) collagen promoter at approximately -1347 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 1(I) collagen OSE2 at -1347 bp reduced the activity of a 1(I) collagen promoter fragment 2-3 fold. Moreover, multimers of this OSE2 at -1347bp confer osteoblast-specific activity to a minimum 1(I) collagen promoter fragment in DNA transfection experiments as well as in transgenic mice. An additional Cbfa1-binding element is present in the 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 1(I) collagen, the mouse 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 this2(I) OSE2 element confer osteoblast-specific activity to the minimum 1(I) collagen promoter in DNA transfection experiments. Thus, our results suggest that Cbfa1 is one of the regulators of the osteoblast-specific expression of both type I collagen genes.
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