癌抑制遺伝子産物の生理機能

• Project/Area Number: 05151072
• Research Category: Grant-in-Aid for Cancer Research
• Allocation Type: Single-year Grants
• Research Institution: National Cancer Center Research Institute and Research Center for Innovative Oncology, National Cancer Center Hospital East
• Principal Investigator: 田矢 洋一 国立がんセンター研究所, 生物, 室長 (60133641)
• Co-Investigator(Kenkyū-buntansha): 原田 久士 阪大, 細胞生体工学センター, 助手 (10222233) ; 神田 忠仁 国立予防衛生研究所, 室長 (60134615) ; 野田 哲生 癌研究所, 部長 (10183550) ; 土田 信夫 東京医科歯科大, 歯, 教授 (60089951) ; 小田 鈎一郎 東京理科大, 基礎工, 教授 (40012736)
• Project Period (FY): 1993
• Project Status: Completed (Fiscal Year 1993)
• Budget Amount: ¥16,500,000 (Direct Cost: ¥16,500,000); Fiscal Year 1993: ¥16,500,000 (Direct Cost: ¥16,500,000)
• Keywords: RB蛋白質 / 細胞周期 / p53 / p300 / IRF / HPVE7 / RBQ-3

Research Abstract

癌抑制RB蛋白質に結合する新しい蛋白質としてクローニングしたRBQ-3のcDNAの全一次構造を決めた。そして、その蛋白質が、発芽酵母の一本鎖DNA結合蛋白質の遺伝子RPA-1のすぐ3側に存在する機能未知の蛋白質と有意な相同性をもつことを見い出した。RBQ-3遺伝子はヒト染色体上の1q32にマップされた。また、核マトリックス蛋白質ラミンAがin vitroでRB蛋白質と結合することを見い出し、その結合に使われるラミンA蛋白質上の領域を同定した。さらに、昨年度に開発したリン酸化ペプチド化学合成法を用いて、RB蛋白質上のリン酸化部位のうちの2カ所に相当するリン酸化ペプチドを合成し、それを抗原として、RB蛋白質上の特定のリン酸化部位のみを認識する抗体の作成に成功した。また、RB結合蛋白質としてクローニングしたRAXが細胞周期の進行を強く阻害することを見い出した。
一方、癌抑制遺伝子産物と予想されるp300がc-jun遺伝子上流の転写制御配列DREに転写因子複合体として結合することを示し、p300の標的遺伝子としてc-junを初めて明らかにした。
p53については、その保存領域とGSTとの融合蛋白質を用いて、これまでに知られているp53結合蛋白質Mdm2やRF-A等とは異なると考えられる複数の新しい結合蛋白質を見い出した。このうち2種類はSV40largeTによって特異的に結合が阻止された。両蛋白質の合成は細胞周期に依存していた。
また一方では、IRF-1が癌抑制遺伝子として白血病発症にも関連することを明らかにした。IRF-1あるいはIRF-2遺伝子欠損マウスも樹立できた。
さらには、G10BPを純度80-90%まで精製し、サウスウエスタン法により、その分子量は約3万と推定した。
APC遺伝子欠損マウスも作成でき、多数の大腸癌の発生が確認できた。
HPV-E6/E7をターゲットとするアンチセンスオリゴヌクレオチドによる子宮頚癌細胞の増殖抑制効果に関しては、オヌクレオチドに修飾基を付加することで顕著な抑制効果が得られた。

Research Products

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