Bone morphogenetic protein-2 induces expression of murine zinc finger transcription factor ZNF450

2004 ◽  
Vol 94 (1) ◽  
pp. 202-215 ◽  
Author(s):  
Alasdair J. Edgar ◽  
Sharon L. Dover ◽  
Melanie N. Lodrick ◽  
Ian J. McKay ◽  
Francis J. Hughes ◽  
...  
2015 ◽  
Vol 291 (3) ◽  
pp. 1148-1161 ◽  
Author(s):  
Chandi C. Mandal ◽  
Falguni Das ◽  
Suthakar Ganapathy ◽  
Stephen E. Harris ◽  
Goutam Ghosh Choudhury ◽  
...  

2000 ◽  
Vol 165 (3) ◽  
pp. 579-586 ◽  
Author(s):  
Y Takazawa ◽  
K Tsuji ◽  
A Nifuji ◽  
H Kurosawa ◽  
Y Ito ◽  
...  

Core-binding factor A1 (Cbfa1), also called Pebp2 alpha A/AML3, is a transcription factor that belongs to the runt-domain gene family. Cbfa1-deficient mice are completely incapable of both endochondral and intramembranous bone formation, indicating that Cbfa1 is indispensable for osteogenesis. Maturation of chondrocytes in these mice is also disorganized, suggesting that Cbfa1 may also play a role in chondrogenesis. The aim of this study was to examine the expression and regulation of Pebp2 alpha A/AML3/Cbfa1 expression in the chondrocyte-like cell line, TC6. Northern blot analysis indicated that Cbfa1 mRNA was constitutively expressed as a 6.3 kb message in TC6 cells and the level of Cbfa1 expression was enhanced by treatment with bone morphogenetic protein-2 (BMP2) in a time- and dose-dependent manner. This effect was blocked by an RNA polymerase inhibitor, 5,6-dichloro-1-beta-d-ribofuranosylbenzimidazole, but not by a protein synthesis inhibitor, cycloheximide. Western blot analysis of the cell lysates using polyclonal antibody raised against Cbfa1 indicated that BMP2 treatment increased the Cbfa1 protein level in TC6 cells. In TC6 cells, BMP2 treatment enhanced expression of alkaline phosphatase and type I collagen mRNAs but suppressed that of type II collagen mRNA. In addition to TC6 cells, Cbfa1 mRNA was also expressed in primary cultures of chondrocytes and BMP2 treatment enhanced Cbfa1 mRNA expression in these cells similarly to its effect on TC6 cells. These data indicate that the Pebp2 alpha A/AML3/Cbfa1 gene is expressed in a chondrocyte-like cell line, TC6, and its expression is enhanced by treatment with BMP.


2006 ◽  
Vol 26 (16) ◽  
pp. 6197-6208 ◽  
Author(s):  
Ming Zhao ◽  
Mei Qiao ◽  
Stephen E. Harris ◽  
Di Chen ◽  
Babatunde O. Oyajobi ◽  
...  

ABSTRACTBone morphogenetic protein 2 (BMP-2) plays a critical role in osteoblast function. InDrosophila, Cubitus interruptus (Ci), which mediates hedgehog signaling, regulates gene expression ofdpp, the ortholog of mammalian BMP-2. Null mutation of the transcription factor Gli2, a mammalian homolog of Ci, results in severe skeletal abnormalities in mice. We hypothesize that Gli2 regulates BMP-2 gene transcription and thus osteoblast differentiation. In the present study, we show that overexpression of Gli2 enhances BMP-2 promoter activity and mRNA expression in osteoblast precursor cells. In contrast, knocking down Gli2 expression by Gli2 small interfering RNA or genetic ablation of the Gli2 gene results in significant inhibition of BMP-2 gene expression in osteoblasts. Promoter analyses, including chromatin immunoprecipitation and electrophoretic mobility shift assays, provided direct evidence that Gli2 physically interacts with the BMP-2 promoter. Functional studies showed that Gli2 is required for osteoblast maturation in a BMP-2-dependent manner. Finally, Sonic hedgehog (Shh) stimulates BMP-2 promoter activity and osteoblast differentiation, and the effects of Shh are mediated by Gli2. Taken together, these results indicate that Gli2 mediates hedgehog signaling in osteoblasts and is a powerful activator of BMP-2 gene expression, which is required in turn for normal osteoblast differentiation.


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