Abstract | BACKGROUND: Idiopathic congenital talipes equinovarus (ICTEV) is a congenital limb deformity. Based on extended transmission disequilibrium testing, Gli-Kruppel family member 3 (Gli3) has been identified as a candidate gene for ICTEV. Here, we verify the role of Gli3 in ICTEV development. METHODS: Using the rat ICTEV model, we analyzed the differences in Gli3 expression levels between model rats and normal control rats. We used luciferase reporter gene assays and ChIP/EMSA assays to analyze the regulatory elements of Gli3. RESULTS: Gli3 showed higher expression levels in ICTEV model rats compared to controls (P < 0.05). We identified repressor and activator regions in the rat Gli3 promoter. The Gli3 promoter also contains two putative Hoxd13 binding sites. Using EMSA, the Hoxd13 binding site 2 was found to directly interact with Hoxd13 in vitro. ChIP assays of the Hoxd13-Gli3 promoter complex from a developing limb confirmed that endogenous Hoxd13 interacts with this region in vivo. CONCLUSION: Our findings suggest that HoxD13 directly interacts with the promoter of Gli3. The increase of Gli3 expression in ICTEV model animal might result from the low expression of HoxD13.
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Authors | DongHua Cao, ChunLian Jin, MeiHong Ren, ChangKun Lin, Xuan Zhang, Ning Zhao |
Journal | BMC musculoskeletal disorders
(BMC Musculoskelet Disord)
Vol. 10
Pg. 142
(Nov 19 2009)
ISSN: 1471-2474 [Electronic] England |
PMID | 19925654
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- GLI3 protein, human
- GLI3 protein, rat
- HOXD13 protein, rat
- Homeodomain Proteins
- Kruppel-Like Transcription Factors
- Nerve Tissue Proteins
- RNA, Messenger
- Transcription Factors
- Zinc Finger Protein Gli3
- Tretinoin
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Topics |
- Animals
- Base Sequence
- Binding Sites
- Cadaver
- Cells, Cultured
- Child
- Child, Preschool
- Chromatin Immunoprecipitation
- Clubfoot
(chemically induced, genetics, metabolism)
- Disease Models, Animal
- Electrophoretic Mobility Shift Assay
- Female
- Gene Expression Regulation, Developmental
- Genes, Reporter
- Hindlimb
(abnormalities, metabolism)
- Homeodomain Proteins
(genetics, metabolism)
- Humans
- Kruppel-Like Transcription Factors
(genetics, metabolism)
- Male
- Molecular Sequence Data
- Nerve Tissue Proteins
(genetics, metabolism)
- Pregnancy
- Promoter Regions, Genetic
- RNA, Messenger
(metabolism)
- Rats
- Rats, Wistar
- Transcription Factors
(genetics, metabolism)
- Transfection
- Tretinoin
- Zinc Finger Protein Gli3
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