| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2006: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2005: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
1 Mouse Polycomb M33(Cbx2) and tri-methylated K27 histoneH3 distribution at fetal gonad specific gene loci.
ChIP analyses using antibody for M33 and tri-methylated K27 histoneH3 in fetal genital ridge lysate were performed.
These factors were tethered around the promoter region of Sox9, Lhx9 genes. Moreover, at the Ad4BP/Sf-1 locus, these factors tethered around the 1st intron, which is 4kb downstream to transcription start site. This region is known as CpG island. These data suggests that, M33 regulate the Ad4BP/Sf-1 locus not only through the modification of the specific histone tail, but also through the DNA methylation process.
2 Mouse Polycomb M33 function through the target region in vivo
To assess the function of M33 at its target region, we established EGFP transgenic mice line, by which we could monitor gene expression levels regulated by the target region. In wild-type fetal gonad, EGFP were activated by Ad4BP/Sf-1 basal promoter connected with 1st exon/intron of Ad4BP/Sf-1 gene. On the other hand, when this construct was in M33KO fetal gonad, EGFP expression was reduced. These data strongly suggests that M33 is indispensable for regulation of Ad4BP/Sf-1 gene expression via its target region on Ad4BP/Sf-1 gene.
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