| Project/Area Number |
06557010
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General medical chemistry
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| Research Institution | Nagoya City University Medical School |
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Principal Investigator |
OKAMOTO Takashi Nagoya City University Medical School Department of Molecular Genetics, Professor, 医学部, 教授 (40146600)
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| Co-Investigator(Kenkyū-buntansha) |
KUGE Shusuke Tokyo University Institute of Medical Science, Department of Microbiology, Resea, 医科学研究所, 助手 (50186376)
KATOH Hiroyuki Hokkaido University School of Pharmacy, Associate Professor, 薬学部, 助教授 (60185866)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥7,800,000 (Direct Cost: ¥7,800,000)
Fiscal Year 1995: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1994: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Transcription Factor / Signal Transduction / NF-kB / thioredoxin / two-hybrid / yAP-1 / USF / Tat / 蛋白・蛋白相互作用 / two-hybrid system / NFkB / PTB / two-hy brid system |
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| Research Abstract |
(1) Redox regulation of NF-kB has been investigated using the following systems.
1) Pathogenetic role of NF-kB in rheumatoid arthritis (RA) has been identified using the synovial fibroblast culture fromthe patients. Co-localization of thioredoxin (TRX) and NF-kB was successfully demonstrated thus confirmed the redox regulation of NF-kB by TRX.
2) Involvement of NF-kB in the cancer cell-endothelial cell adhesion was further investigated. Anti-oxidants were demonstrated to be effective in blocking NF-kB activation and cancer cell adhesion.
3) We have established a recombinant cell line survival of which is under the control of NF-kB.This cell line has been shown to be applicable for the screening of novel anti-NF-kB compounds.
(2) Activity of S.cerevisiae yAP-1, an AP-1-like transcription factor, is elevated by oxidation stress, and yAP-1 activates transcription from an AP-1 dependent reporter gene and TRX2 that is required for resistance to peroxidation. Furthemore yAP-1 migrates into nucleus upon oxidation stress and this change in localization is important for transcriptional regulation. Cystein residues in the essential C-terminal region are involved in this migration.
(3) A bZip protein Fral has been isolated by the two-hybrid screening for protein specifically interacting with bHLHZip protein USF.Both of bZip and N terminal regions are required for this interaction. Expression of exogenous USF caused down-regulation of AP1 activity, which suggests that USF is a negative regulator of AP1. We have also isolated cDNA for a novel protein TAM2 that interacts with the activation region of HIV-1 Tat by the two-hybrid system. Since TAM2 pulled down both cdk7 and cyclinH,we suggest that Tat recruits TFIIH via TAM2 and activates initiation and elongation of transcription.
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