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Published 2 September 2003. doi:10.1083/jcb.200211089
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© The Rockefeller University Press, 0021-9525/2003/9/833 $5.00
The Journal of Cell Biology, Volume 162, Number 5, 833-842

Article

Type X collagen gene regulation by Runx2 contributes directly to its hypertrophic chondrocyte–specific expression in vivo



Qiping Zheng1, Guang Zhou1, Roy Morello1, Yuqing Chen1, Xavier Garcia-Rojas1 and Brendan Lee1,2

1 Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030
2 Howard Hughes Medical Institute, Baylor College of Medicine, Houston, TX 77030

Address correspondence to Brendan Lee, Dept. of Molecular and Human Genetics Baylor College of Medicine One Baylor Plaza, Rm 635E, Houston, TX 77030. Tel.: (713) 798-8835. Fax: (713) 798-5168. email: blee{at}bcm.tmc.edu

The {alpha}1(X) collagen gene (Col10a1) is the only known hypertrophic chondrocyte–specific molecular marker. Until recently, few transcriptional factors specifying its tissue-specific expression have been identified. We show here that a 4-kb murine Col10a1 promoter can drive ß-galactosidase expression in lower hypertrophic chondrocytes in transgenic mice. Comparative genomic analysis revealed multiple Runx2 (Runt domain transcription factor) binding sites within the proximal human, mouse, and chick Col10a1 promoters. In vitro transfection studies and chromatin immunoprecipitation analysis using hypertrophic MCT cells showed that Runx2 contributes to the transactivation of this promoter via its conserved Runx2 binding sites. When the 4-kb Col10a1 promoter transgene was bred onto a Runx2+/- background, the reporter was expressed at lower levels. Moreover, decreased Col10a1 expression and altered chondrocyte hypertrophy was also observed in Runx2 heterozygote mice, whereas Col10a1 was barely detectable in Runx2-null mice. Together, these data suggest that Col10a1 is a direct transcriptional target of Runx2 during chondrogenesis.

Key Words: CBFA1; COL1OA1; CCD; MCT cells; transgenic mice

Abbreviations used in this paper: CCD, cleidocranial dysplasia; EMSA, electrophoretic mobility shift assays; P1, postnatal day 1; SMCD, Schmid metaphyseal chondrodysplasia.


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