| Project/Area Number |
06454604
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Biological pharmacy
|
|---|
| Research Institution | The Institute of Physical and Chemical Research (RIKEN) |
|---|
Principal Investigator |
YOKOYAMA Kazushige Gene Bank, Vice-researcher, ジーンバンク室, 副主任研究員 (80182707)
|
|---|
| Project Period (FY) |
1994 – 1996
|
| Project Status |
Completed (Fiscal Year 1996)
|
| Budget Amount *help |
¥5,900,000 (Direct Cost: ¥5,900,000)
Fiscal Year 1996: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1995: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1994: ¥2,800,000 (Direct Cost: ¥2,800,000)
|
|---|
| Keywords | Adenovirus / MHC class I / EIA / TBP / Negative repression / p300 / MAZ / HMG 1 / アデノフィルス / E1A / P300 / ドメイン解析 / アデノウィルス / 負の転写制御 / E1Aのドメイン構造 |
|---|
| Research Abstract |
The primary rodent cells transformed by the oncogenic Ad12 virs expressed reduced amount of class I antigen as opposed to cells transformed by the non-oncogenic Ad5 (2) virs. We study here the mechanism of the transcriptional repression of H-2K^b gene by Ad12-EIA.The promoter assay with the successive 5' end delection and substitution CAT constructs demonstrated the negative regulatory elements were localized in the distal 316 bp sequences (-1835 to 1521). The cell-surface expression of MHC class I cDNA expresssion vector, in which the (CAA) n repeats in the 316 bp region was inserted, as compared with the mutated (CAA) n sequence. The (CAA) n repeats can be contributed to the EIA mediated-negative reperession. These nuclear factors-binding to the negative element has been purified and we isolated the cDNA clones encoded these proteins bound to (CAA) n repeated elements, MAZ (Myc-associated zinc finger protein) and HMG-1. Furthermore, the encoded proteins, which bind to this repeats, were shown to associate with the EIA binding cellular protein, p300 in Ad12-EIA.Second element is the TATA-like sequence located in the far-upstream region. The TBP (or TBP like molecules) can bind to this upstream TATA-like and regular TATA box in a competition binding manner of each site. The p300 cellular protein can also bind this TBP in Ad-EIA.Ths we conclude treat the ELA,MAZ,HMGl and TBP protein complex are directly or indirectly involved in the EIA mediated suppression of MHC class I gene. The EIA protein released the p300 pretein from these complex, to result the negative expression on the MHC class I gene.The protein-protein interaction between these proteins and EIA (condluding p300) was analyzed in vitro and in vivo to study the controlled mechanism of the EIA mediated repression of H-2K^b gene by Ad12-EIA.
|
