エンドセリン受容体の遺伝子転写調節機構と細胞増殖・接着における意義に関する研究

• Project/Area Number: 07671130
• Research Category: Grant-in-Aid for General Scientific Research (C)
• Allocation Type: Single-year Grants
• Research Field: 内分泌・代謝学
• Research Institution: Kyoto University
• Principal Investigator: 吉政 孝明 京都大学, 医学研究科, 助手 (00252429)
• Co-Investigator(Kenkyū-buntansha): 荒井 宏司 京都大学, 医学研究科, 助手 (00263088)
• Project Period (FY): 1995
• Project Status: Completed (Fiscal Year 1995)
• Budget Amount: ¥2,100,000 (Direct Cost: ¥2,100,000); Fiscal Year 1995: ¥2,100,000 (Direct Cost: ¥2,100,000)
• Keywords: エンドセリン / エンドセリン受容体 / 転写 / シス配列 / RNAスプライシング

Research Abstract

1.エンドセリン(ET)受容体遺伝子の転写調節機構の解析:
ヒトET-A受容体遺伝子5′隣接領域のfunctional assayを用いた検討により、-0.2kb〜-857b間に負の転写調節領域の、-137b〜-53b間及び-53b〜+251b間に正の転写調節領域の存在を明らかにした。更に、-137b〜-53bの正の転写調節領域をA7r5核抽出物を用いたゲルシャフトアッセイより解析した結果、-106b〜-78b間に核蛋白の結合を認めた。フットプリントアッセイの結果、-91b〜-83b間にCCCCACCTTからなる核蛋白結合部位を認めた。このようにET-A受容体遺伝子のbasal transcriptional activityには新しいシス配列が関与していることが示唆された。
2.エンドセリン-A受容体遺伝子alternative RNA splicingの遺伝子発現調節機構における関与の検討:
ヒト肺total RNAをET-A受容体cDNAの全翻訳領域を増幅するプライマーを用いたRT-PCRを行うと、予想されるET-A受容体転写産物以外にbandsを認めた。これらに対するクローンをクローニングすると、それぞれエクソン4あるいはエクソン3,4がsplice outされたアイソホームであった。ET-A受容体遺伝子発現調節機構の一つを示唆するものと思われた。

Research Products

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