1994 Fiscal Year Final Research Report Summary
Analysis of enhancer/silencer regions of TCR gene
| Project/Area Number |
05670293
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Immunology
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| Research Institution | Chiba University |
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Principal Investigator |
KANNO Masamoto School of Med.Chiba Univ.Assoc.Prof., 医学部, 助教授 (40161393)
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| Co-Investigator(Kenkyū-buntansha) |
TANIGUCHI Masaru School of Med.Chiba Univ.Prof., 医学部, 教授 (80110310)
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| Project Period (FY) |
1993 – 1994
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| Keywords | TCR / ets / enhancer |
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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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