| Project/Area Number |
11680685
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Molecular biology
|
|---|
| Research Institution | Saitama Medical School |
|---|
Principal Investigator |
HISATAKE Koji Saitama Medical School, Biochemistry, Associate Professor, 医学部, 助教授 (70271236)
|
|---|
| Project Period (FY) |
1999 – 2000
|
| Project Status |
Completed (Fiscal Year 2000)
|
| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
|
|---|
| Keywords | General Transcription Factor / Baculo virus / Protein Phosphorylation / Transcription / TFIIH / Capping Reaction / バキュウロウイル / リン酸化 / 伸長反応 / CTD / バキュロウイルス |
|---|
| Research Abstract |
1. Role of ERCC3 in transcriptional activation by GAL4-VP16
In basal transcription, ERCC3 helicase acts during promoter opening and promoter escape. We tested at which step ERCC3 helicase acts during transcriptional activation by GAL4-VP16. The effect of TFIIH mutants were first tested using premelted templates, and revealed that TFIIH functions during promoter escape in activated transcription. The effect is specific to activation. Using the various mutant templates, we found that activators act during a very early stage of transcription. The effect requires coactivator PC4. RNA polymerase II is unstable during the early phase of transcription and may abort transcription. With the use of ATP analogs, we found that RNA polymerase II may detach from the template when ERCC3 helicase activity is blocked. Taken together, transcriptional activation involves the enhancement of recruitment of transcriptional machinery and promoter escape. Rate constant of promtoer escape is not stimulated in activated transcription.
2. Role of TFIIH in transcription-coupled capping reactions
In collaborated with Dr.Mizumoto, we have developed a system in which transcription and capping reactions can occur together. The system includes recombinant factors except RNA polymerase II.In this system, capping reaction occurs simultaneously with transcription initiation. The molecular analyses of capping and its relation to transcription initiation are in progress.
|
