| Project/Area Number |
04454271
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Jichi Medical School |
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Principal Investigator |
NAGANO Kei Jichi Med.Sch., Dept.Biol., Prof., 医学部, 教授 (70048940)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAKAMI Kiyosi Jichi Med.Sch., Dept.Biol., Assoc.Prof., 医学部, 助教授 (54191016)
INOHARA Naohiro Jichi Med.Sch., Dept.Biol., Research Associate
猪原 直弘 自治医科大学, 医学部・生化学, 助手 (60232576)
鈴木 由利子 自治医科大学, 医学部, 助手 (50211610)
川上 潔 自治医科大学, 医学部, 助教授 (10161283)
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| Project Period (FY) |
1992 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1994: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1993: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1992: ¥3,700,000 (Direct Cost: ¥3,700,000)
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| Keywords | Na, K-ATPase / Ion homeostasis / Gene expression / Transcription factor / Subunit / Active transport / Ion pump / 転写制御 / alphaサブユニット / betaサブユニット / 制御領域 / 制御因子 / Sp1 / E-box / ナトリウムポンプ / 筋細胞 / 遺伝子発現制御 / Eボックス / DNaseIフットプリント法 / メチル化干渉法 |
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| Research Abstract |
Na, K-ATPase alpha and beta subunit isoform genes were studied for their molecular apparatus of expression regulation : 5' upstream cis-elements and the corresponding trans-acting factors. Isoform genes used were house-keeping alpha1 subunit gene, alpha2 subunit gene expressed mainly in muscle cells, and beta2 subunit gene whose product is also known as AMOG (adhesion molecule on glig).
alpha1 : A regulatig region (ARE=alpha1 gene regulation element) was found at -102--61. At least six kinds of protein binding factors interact at the ARE region. Several factors are already known to regulate the expression of other genes (HEB and ATF). A new factor, AREB6, was obtained by south-western cloning. AREB6 is a Zn-finger protein. The seven fingers are separated into two clusters of three and four, and located in the N-and C-halves of the molecule, respectively. Up-or down-regulating effects of these elements were different according to different cells transfected, indicating complex accomodation of Na, K-ATPase expression regulation.
alpha2 : Two Sp1-binding and a similar(GGGAGG)sequences and two E-boxes are found at -175--108. Several fold expression activation by this region was observed in the transfection to the myoblast-derived L6 cells.
beta2 : The specific element, AMRE (AMOG regulating element), which is an expression regulating region in rat neuroblastoma-derived 103B cells, was also shown to regulate Na, K-ATPase gene expression in cultured rat astro-cyte cells as well as other cells from several other tissues. A trans-acting factor to AMRE was found to be competed by an Sp1-binding sequence. The binding was also inhibited by anti-Sp1 antibody.
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