| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Research Abstract |
The TEF-1 family of transcription factors consists of at least four members, Tead1/TEF-1, Tead2/ETF, Tead3/ETFR-1 and Tead4/ETFR-2, containing a highly conserved TEA DNA-binding domain, and have been implicated in cardiovascular development and function. It has recently been reported that the scalloped (Sd) protein, a Drosophila homologue of TEF-1, controls the gene expression involved in the wing development through protein-protein interaction with Vestigial (Vg). We have isolated a mouse cDNA encoding amino acid sequences similar to those of the Sd-binding domain (SdBD) of Vg by RTPCR and RACE methods. The protein deduced from the cDNA, termed mouse Vg-related Factor (mVRF), consists of 307 amino acid residues. The amino acid sequence of mVRF showed 49 % identity to that of Tondu, a recently identified human counterpart of Vg, but diverged from that of Drosophila Vg except for that of SdBD. Northern blot analysis revealed that mVRF mRNA expressed in placenta, embryo and testis. Transient transfection assay using the luciferase reporter gene and mVRF expression vector suggested that overexpression of mVRF represses the TEF-1-dependent transcriptional activity. On the other hand, we have identified and characterized the cell-specific 117-bp enhancer sequence in the first intron of the mouse ETF/Tead2 gene required for transcriptional activation in ETF/Tead2 gene expressing cells, such as P19 cells. We have also identified one GC-rich sequence (the GC box) and two tandemly repeated GA-rich sequences (the proximal and distal GA elements) in the enhancer and characterized their possible binding proteins, which may play a role in the cell-specific enhancer function.
|
