癌抑制遺伝子産物の生理機能

• Project/Area Number: 06280124
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• 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, 生物学部, 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) ; 松七五三 仁 東大, 医科研, 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, 歯, 教授 (60089951) ; 小田 鈎一郎 東京理科大, 基礎工, 教授 (40012736)
• Project Period (FY): 1994
• Project Status: Completed (Fiscal Year 1994)
• Budget Amount: ¥16,500,000 (Direct Cost: ¥16,500,000); Fiscal Year 1994: ¥16,500,000 (Direct Cost: ¥16,500,000)
• Keywords: 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

Research Abstract

RB蛋白質をリン酸化するRBキナーゼについては、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。さらに、RB蛋白質上にはサイクリンD1-Cdk4でリン酸化されるけれども他のサイクリンEやA依存性キナーゼではリン酸化されない部位が存在することも見い出した。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。一方では、以前に開発したリン酸化ペプチドの化学合成法を用いて、RB蛋白質上の3ヵ所のリン酸化部位をそれぞれ認識する抗体の作製に成功した。これには、上述のサイクリンD1-Cdk4特異的なリン酸化部位も含まれる。
また、接触阻止においてはサイクリンD依存性キナーゼ活性が阻害されるが、その際、サイクリンDとCdk4は複合体を形成しているのに、Cdk4の活性化には必須であるThr^<172>がリン酸化されていないことを見い出した。しかし、この部位をリン酸化する酵素であるCdk活性化キナーゼ(CAK)の活性には大きな変化はなかった。一方、_p27^<kip1>が多量に存在することで、CAKがCdk4を基質として認識するのを阻害し、結果として、接触阻止ではサイクリンD依存性キナーゼの活性化が阻害されていることを明らかにした。
p53については、ヒトのp53温度感受性変異株(バリン138)を作製した。この変異株をラット胎児繊維芽細胞に活性化rasとともに導入した細胞では、waf1発現の上昇を経ずにG1停止が起きること、Saos-2に導入した細胞ではG1とG2/Mの2つの時期で停止が起きることが示唆された。
また、ショウジョウバエの癌抑制遺伝子の1(2)glのマウスホモログmgl-1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結果、mgl-1を欠損したマウスでは胎生期の神経上皮細胞の異常増殖が起きることが判明したが、腫瘍の発生は見られなかった。

Research Products

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