TY - JOUR
T1 - A novel cis-element that enhances connective tissue growth factor gene expression in chondrocytic cells
AU - Eguchi, Takanori
AU - Kubota, Satoshi
AU - Kondo, Seiji
AU - Kuboki, Takuo
AU - Yatani, Hirofumi
AU - Takigawa, Masaharu
N1 - Funding Information:
The authors thank Drs. Tohru Nakanishi, Takashi Nishida, Gen Yosimichi, Norifumi H. Moritani, Yoshiki Mukudai, Eiji Nakata, and Kumiko Nawachi for helpful suggestions, Ms. Kazumi Ohyama for technical assistance, and Ms. Yuki Nonami and Ayako Shizuku for secretarial assistance. This work was supported in part by grants-in-aid for Scientific Research (S.K., M.T.) and Exploratory Research (M.T.) from the Ministry of Education, Science, Sports, and Culture of Japan, Grants in Aid for Scientific Diseases of the Ministry of Public Health and Welfare of Japan (M.T.), and by grants from the Nakatomi Health Science Foundation (S.K.), the Sumitomo Foundation (M.T.) and the Ryobi-teien Memorial Foundation (S.K.).
PY - 2002
Y1 - 2002
N2 - To clarify the chondrocyte-specific regulatory mechanism of connective tissue growth factor (ctgf) gene expression, we analyzed the functionality and DNA-protein interaction of the CTGF promoter. Comparative luciferase assay of the CTGF promoter deletion mutants among HCS-2/8 chondrocytic cells and fibroblastic cells revealed that a 110-bp region in the promoter was crucial for the HCS-2/8-specific transcriptional enhancement. Subsequent competitive gel shift assay revealed that transcription factors in HCS-2/8 nuclei bound to a 60-bp portion in the corresponding region. Relative luciferase activity from a CTGF promoter with mutant TGF-β response element (TbRE) was 16.9% lower than that from an intact promoter. On the other hand, relative luciferase activity from a CTGF promoter with 4 bp point mutations at 30 bp upstream of the TbRE was 47.7% lower than that from the intact one. The binding activity of HCS-2/8 nuclear factor(s) to the sequence over the 4-bp was remarkably higher than that of any nuclear extract from other types of cells. Therefore, we entitled the sequence ‘TRENDIC’, a transcription enhancer dominant in chondrocytes, which stands for a novel enhancer for chondrocyte-specific CTGF gene expression.
AB - To clarify the chondrocyte-specific regulatory mechanism of connective tissue growth factor (ctgf) gene expression, we analyzed the functionality and DNA-protein interaction of the CTGF promoter. Comparative luciferase assay of the CTGF promoter deletion mutants among HCS-2/8 chondrocytic cells and fibroblastic cells revealed that a 110-bp region in the promoter was crucial for the HCS-2/8-specific transcriptional enhancement. Subsequent competitive gel shift assay revealed that transcription factors in HCS-2/8 nuclei bound to a 60-bp portion in the corresponding region. Relative luciferase activity from a CTGF promoter with mutant TGF-β response element (TbRE) was 16.9% lower than that from an intact promoter. On the other hand, relative luciferase activity from a CTGF promoter with 4 bp point mutations at 30 bp upstream of the TbRE was 47.7% lower than that from the intact one. The binding activity of HCS-2/8 nuclear factor(s) to the sequence over the 4-bp was remarkably higher than that of any nuclear extract from other types of cells. Therefore, we entitled the sequence ‘TRENDIC’, a transcription enhancer dominant in chondrocytes, which stands for a novel enhancer for chondrocyte-specific CTGF gene expression.
KW - Chondrocyte
KW - Connective tissue growth factor
KW - Gene expression
KW - Transcription factor
KW - Transforming growth factor-β
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U2 - 10.1016/S0006-291X(02)00700-3
DO - 10.1016/S0006-291X(02)00700-3
M3 - Article
C2 - 12150969
AN - SCOPUS:0036062468
SN - 0006-291X
VL - 295
SP - 445
EP - 451
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -