| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2005: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
|
|---|
| Research Abstract |
We previously developed a novel functional complementation screening in yeast. Using the system, we tried to verify the potential functions of active-H-ras and tumor suppressor gene menin. We screened a yeast mutant library and isolated one yeast mutant which shows dependency of active-H-ras for its cell growth. However, the phenotype of the mutant did not stable and many revertants were appeared from the mutant under the restricted condition. Therefore, we next used modified yeast two-hybrid systems, in which human genes stably express, to search the potential function of human genes. Using the systems, we searched associated proteins with the U1 small nuclear ribonucleoprotein (snRNP)-specific protein U1C, since aberrant splicing was frequently observed in cancer cells and snRNPs play an essential role in splicing through a large complex known as the spliceosome. We identified one isoform of coactivator-associated arginine methyltransferase 1 (named CARM1-v3) as a U1C interacting protein. It has been shown to act as a transcriptional coactivator to enhance transcriptional activation by nuclear receptors, and affects 5' splice site selection of the pre-mRNA splicing. CARM1-v3, but not the other isoforms, strongly stimulates a shift to the distal 5'-splice site of the pre-mRNA when the adenoviral E1A minigene is used as a reporter, and enhances the exon skips in the CD44 reporter. These results suggest that the transcriptional coactivator can affect the splice site decision, and may participate in the aberrant splicing in cancer cells.
|
