癌抑制遺伝子産物の生理機能

• 研究課題/領域番号: 07272110
• 研究種目: 重点領域研究
• 配分区分: 補助金
• 研究機関: 国立がんセンター
• 研究代表者: 田矢 洋一 国立がんセンター, 研究所生物学部, 室長 (60133641)
• 研究分担者: 神田 忠仁 国立予防衛生研究所, 室長 (60134615) ; 松七五三 仁 東京大学, 医科学研究所, 助教授 (50251442) ; 野田 哲生 癌研究所, 部長 (10183550) ; 土田 信夫 東京医科歯科大学, 歯学部, 教授 (60089951) ; 小田 鈎一郎 東京理科大学, 基礎工学部, 教授 (40012736)
• 研究期間 (年度): 1995
• 研究課題ステータス: 完了 (1995年度)
• 配分額: 17,300千円 (直接経費: 17,300千円); 1995年度: 17,300千円 (直接経費: 17,300千円)
• キーワード: RB蛋白質 / p53 / 癌抑制遺伝子 / サイクリンD / Cdk4 / Cdkインヒビター / mgl-1 / ヒトパピローマウィルス

研究概要

1.酵母のTwo-hybrid系を利用して、HPV16のE6蛋白と複合体を形成する2つの細胞蛋白を得た。1つは、HUMHMCM2(酵母MCM2のヒトホモログ)であり、他の1つは未知の蛋白(約60Kd)であった。
2.サイクリンD1-Cdk4が、他のサイクリンEやA依存性キナーゼとは異なったアミノ酸配列の所をリン酸化することを初めて明らかにした。サイクリンD1は癌遺伝子にもなるサイクリンであるので、この発見によって、サイクリンD1が細胞癌化に関与するメカニズムの解明が大きく進展すると期待できる。
3.Drosophilaの癌抑制遺伝子l(2)glのマウスホモログmgl-1ならびにmgl-2遺伝子の機能をジーンターゲッティングにより解析した。mgl-1ホモ欠損マウスでは水頭症を呈し、出生後48時間以内に死亡し、組織学的には神経上皮細胞の異常が認められた。一方、mgl-2ホモ欠損マウスは、ほとんどが生殖能力を有する生体となるが、すべて矮小であった。
4.p21の変異蛋白を複数作製して精製し、アミノ酸置換されたp21はサイクリンDl/Cdk4複合体に結合するがその酵素活性を阻害できないことを見い出した。この変異p21をp16とともにヒト細胞に発現させると、p16に拮抗的な変異p21の作用が観察された。
5.ラットcdc2プロモーターのGl/S境界での活性化に関与するエンハンサーAAGTTACAAATAに結合する蛋白因子cdc2E1とcdc2E2のcDNAを単離した結果、E1はSRE中のCArGbox結合因子をコードし、E2は未知の遺伝子であることがわかった。

研究成果

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