| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1993: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Research Abstract |
It has been known that TCR-alpha enhancer is subdivided to Talpha1-4. In our recent study we demonstrate several major developments : (1) Talpha2 region is most critical to exert enhancer function. This Talpha2 region has reported to have a binding sites for TCF-1/LEF-1, Talpha2B and Ets-1. (2) By our in vivo analysis, 5'-CATCCTC-3' sequence is very important for enhancer function, which is overlapped with reported Ets-1 binding site. Moreover, by in vitro analysis, we identified at least three different Ets-1 related proteins are interacting with this Talpha2 region and, especially, one of them seems to have Ets-1 like binding specificity to 5'-ATCC-3' sequence. However none of these proteins seems to have a character of reported Ets-1 nor reactivity to anti-Est-1 and anti-Ets-2 monoclonal antibodies by gel-shift assay. Our present dats is the first case to demonstrate the binding of several endogenous (natural) Ets-1 family proteins to the Talpha2 region.
To determin the molecular weigth of the proteins, we performed UV-cross link experoment. It indicates novel 10kDa protein is binding to Talpha2 ets site with weakly associated 31kDa protein. By 3'-RACE RT-PCR,we islated a partial cDNA fragment with degenerated ets domain oligonucleotides. This cDNA frament sequence reveal that conserved ets domain with novel c-terminal half of new protein. We are currently trying to isolate full-length cDNA.
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