Runx2 contributes to murine Col10a1 gene regulation through direct interaction with its cis-enhancer
| Autor: | Yuqing Chen, Brendan Lee, Dobrawa Napierala, Siying Wang, Qiping Zheng, Sam Abbassi, Yaojuan Lu, Feifei Li, Xiangyun Duan, Ming-de Ding |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Col10a1
Endocrinology Diabetes and Metabolism Transgene Molecular Sequence Data lac operon Core Binding Factor Alpha 1 Subunit Mice Transgenic Biology 03 medical and health sciences Mice 0302 clinical medicine cis-enhancer Runx2 Genes Reporter Animals Orthopedics and Sports Medicine Electrophoretic mobility shift assay Enhancer Promoter Regions Genetic Gene Base Pairing 030304 developmental biology Regulation of gene expression 0303 health sciences Binding Sites Base Sequence hypertrophic chondrocytes transgenic studies Transfection Original Articles DNA Molecular biology Enhancer Elements Genetic Gene Expression Regulation Tandem Repeat Sequences 030220 oncology & carcinogenesis Mutation Chromatin immunoprecipitation Collagen Type X Protein Binding |
| Zdroj: | Journal of Bone and Mineral Research |
| ISSN: | 1523-4681 0884-0431 |
| Popis: | We have recently shown that a 150-bp Col10a1 distal promoter (−4296 to −4147 bp) is sufficient to direct hypertrophic chondrocyte-specific reporter (LacZ) expression in vivo. More recently, through detailed sequence analysis we identified two putative tandem-repeat Runx2 binding sites within the 3′-end of this 150-bp region (TGTGGG-TGTGGC, −4187 to −4176 bp). Candidate electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation, and transfection studies demonstrate that these putative Runx2 sites bind Runx2 and mediate upregulated Col10a1/reporter activity in vitro. Transgenic studies using the 5′-sequence without Runx2 sites were not able to drive the cell-specific LacZ reporter activity, suggesting the in vivo requirement of the Runx2 sites located in the 3′-end in mediating Col10a1/reporter expression. Indeed, mutating the Runx2 sites in the context of the 150-bp promoter abolishes its capacity to drive hypertrophic chondrocyte-specific reporter expression in transgenic mice. We have also generated multiple transgenic mouse lines using only the 3′-sequence containing the Runx2 sites to drive the LacZ gene. Interestingly, no hypertrophic chondrocyte-specific blue staining was observed in these transgenic mice. Together, our data support that Runx2 directly interacts with murine Col10a1 cis-enhancer. This interaction is required but not sufficient for cell-specific Col10a1 promoter activity in vivo. Additional cooperative/repressive elements within the 5′- or 3′-sequences of this 150-bp promoter are needed to work with Runx2 together to mediate cell-specific Col10a1 expression. Further delineation of these elements/factors has the potential to identify novel therapeutic targets for multiple skeletal disorders, including osteoarthritis, that show abnormal Col10a1 expression and altered chondrocyte maturation. © 2011 American Society for Bone and Mineral Research |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text