| Budget Amount *help |
¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Research Abstract |
We isolated a 2.8 kb cDNA clone encoding a 436-amino acid protein that dictates tissue-specific expression of human aromatase from a human placenta! cDNA library using a yeast one-hybrid system with a trophoblast-specific element 2 as a reporter sequence. The protein was revealed to be identical to hGCMa, a mammalian homologue of the Drosophila GCM (glia cells missing). Drosophila GCM is a neural transcription factor that regulates glial cell fate.
Expression of hGCMa is restricted to the placenta in human. The protein expressed in Sf 21 cells also binds to PLE1 in the leptin promoter among other cis-elements reported to confer placenta-specific expression, suggesting that hGCMa is a placenta-specific transcription regulator, possibly involved in the expression of multiple placenta-specific genes (Yamada K., Ogawa H., et al. 1999 J. Biol. Chem. 274: 32279-32286).
We also determined the genomic organization, chromosomal localization, and the complete 22 kb DNA sequence of hGCMa gene. It contains six exons and five introns, encoding a 2.8 kb cDNA. Overall genomic organization is similar for the human and mouse. Several potential binding sites for transcription factors like GATA, Oct-1, and βHLH proteins were found in the 5'-flanking region of the human gene. A DNA motif for GCM protein binding exists in the 5'-flanking region that is highly homologous with that of the mouse gene. The location of this gene was mapped to chromosome 6 using fluorescence in situ hybridization (Yamada K., Ogawa H., et al. 2000 Biochem. Biophys. Res. Commun. 278: 134-139).
We prepared anti-hGCMa antibody in rabbits. In preliminary experiments, it reacted with also several human blood cells besides the placenta. Now we are going to prepare more specific antibody against hGCMa.
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