| Project/Area Number |
11557031
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
Hygiene
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
SAKAI Toshiyuki Kyoto Prefectural University of Medicine, Department of Preventive Medicine, Professor, 医学部, 教授 (20186993)
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| Co-Investigator(Kenkyū-buntansha) |
NOMURA Hitoshi Chugai Pharmaceutical Co., Ltd. Takada Research Lab. Senior Researcher, 高田研究所, 創薬研究担当部長
YOSHIDA Tatsushi Kyoto Prefectural University of Medicine, Department of Preventive Medicine, Assistant Professor, 医学部, 助手 (80315936)
SOWA Yoshihiro Kyoto Prefectural University of Medicine, Department of Preventive Medicine, Lecturer, 医学部, 講師 (70315935)
FUJII Hideji Nippon Kayaku Co., Ltd. Parmaceutical Group. Drug Research Department. Vice-eniro Researcher, 制癌剤創薬部門, 副主任研究員
MIZUNO Takakazu Chugai Pharmaceutical Co., Ltd. Fuji Gotemba Research Lab. Senior Researcher, 富士御殿場研究所, リーダー
増田 悦子 山之内製薬株式会社, 分子医学研究所, 主管研究員
高橋 千絵 京都府立医科大学, 医学部, 助手 (90305574)
加藤 満雄 京都府立医科大学, 医学部, 講師 (70260221)
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| Project Period (FY) |
1999 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥13,400,000 (Direct Cost: ¥13,400,000)
Fiscal Year 2002: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2001: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 1999: ¥3,200,000 (Direct Cost: ¥3,200,000)
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| Keywords | gadd45 / uv / promoter / p53 / TSA / Oct-1 / NF-Y / luciferase / プロモーター / ケルセチン / フラボノイド / HeLa / ルテオリン / アピゲニン / 酪酸 / CCAAT / 転写因子 / UV / 遺伝子修復 / Oct1 / p27 / Vitamin D3 / WAF1 / Sp3 |
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| Research Abstract |
The gadd45 gene is transcriptionally activated by UV irradiation through p53-dependent or p53-independent pathways. To investigate the sequences involved in induction of gadd45 by UV irradiation in a p53-independent pathway, we performed mutation analyses of the human gadd45 promoter fused to the luciferase reporter gene in cell lines in which p53 was inactivated. We found that the UV-responsive element was involved in the Oct-1 binding site at -99 bp relative to the transcription start site. Electrophoretic mobility shift assays showed that Oct-1, a transcription factor, bound this element on the gadd45 gene, although the intensity and mobility pattern of the retarded bands were not altered by UV irradiation. These results suggest that the Oct-1 regulatory element might be one of the essential elements involved in the activation of the gadd45 promoter by UV irradiation in a p53-indepeadent pathway.
Histone deacetylase (HDAC) inhibitors have been revealed to cause growth arrest at G1 and /or G2/M phases, differentiation, and/or apoptosis in a wide variety of tumor cells. We demonstrated that gadd45, which has been shown to cause cell cycle arrest at G2/M phase and take part in genotoxic stress-induced apoptosis, is also induced by a typical HDAC inhibitor, trichostatin A (TSA), through its promoter in a p53-independent manner. To identify the mechanism of activation of the gadd45 promoter, we performed luciferase reporter analyses and elecrophoretic mobility shift assays. Analysis of the gadd45 promoter revealed that both the Oct-1 and CCAAT sites are needed for the full activation by TSA. We also found that transcription factors, Oct-1 and NF-Y, specifically bind to each site. Our results suggest that HDAC inhibitors can induce gadd4S through its promoter without functional p53 and both the Oct-1 and NF-Y concertedly participate in TSA-induced activation of the gadd45 promoter.
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