Analysis of enhancer/silencer regions of TCR gene
Project/Area Number |
05670293
|
Research Category |
Grant-in-Aid for General Scientific Research (C)
|
Allocation Type | Single-year Grants |
Research Field |
Immunology
|
Research Institution | Chiba University |
Principal Investigator |
KANNO Masamoto School of Med.Chiba Univ.Assoc.Prof., 医学部, 助教授 (40161393)
|
Co-Investigator(Kenkyū-buntansha) |
TANIGUCHI Masaru School of Med.Chiba Univ.Prof., 医学部, 教授 (80110310)
|
Project Period (FY) |
1993 – 1994
|
Project Status |
Completed (Fiscal Year 1994)
|
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)
|
Keywords | TCR / ets / enhancer / T細胞抗原受容体 / 転写因子 / ets遺伝子 |
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.
|
Report
(3 results)
Research Products
(9 results)