| Project/Area Number |
06833008
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
老化(加齢)
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
KUMAZAKI Tsutomu Research Institute for Radiation Biology and Medicine, Department of Biochemsitry and Biophysics, Research Associate Hiroshima University, 原爆放射能医学研究所, 助手 (20161698)
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| Co-Investigator(Kenkyū-buntansha) |
HAMADA Katsutomo Research Institute for Radiation Biology and Medicine, Research Associate Hirosh, 原爆放射能医学研究所, 助手 (00136144)
MITSUI Youji National Institute of BioScience and Human Technology, Chief Senior Scientist, 生命工学工業技術研究所, 首席研究官
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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 |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | Cellular aging / Endothelial cells / Fibronectin / Transcription factors / Gel shift assay / Supershift assay / 遺伝子発現制御 / 線維芽細胞 |
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| Research Abstract |
Previously, we showed that the expression of the fibronectin (FN) gene is enhanced during aging of human endothelial cells and fibroblasts. To elucidate the mechanism, we explored binding proteins to the FN promoter. The promoter contains sites for the general transcription factors : CAAT-binding transcription factor (CTF), promoter-specific transcription factor-1 (Sp1), and transcription factor-IID (TFIID). The promoter also contains sites for inducible transcription factors, cAMP-responsive element (CRE) -binding protein (CREB), and Activator protein 2 (AP-2). We synthesized ten different oligonucleotides for these and other potential transcription factor-binding sites. Using these oligonucleotides, we searched for binding proteins in young and old endothelial cells by electrophoretic mobility shift and supershift assays. Our results showed that AP-1 decreased with aging, but Sp1 and CREB1 were unaffected. However, decreased binding activities to CRE at positions-170 and -415 were shown in old cells. This could be explained by the decrease of AP-1 because these CREs bound not only CREB1 but also AP-1. Moreover, we observed that the binding activities of TFIID,CTF,and binding proteins to -40, -120 and -260 regions increased. These differential changes may cause the enhancement of FN expression in senescent cells.
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