A study on the mechanism of PI3K/AKT pathway activation by v-Crk oncogene

v-Crk癌基因激活PI3K/AKT通路机制的研究

• 批准号: 13670308
• 负责人: AKAGI Tsuyoshi
• 金额: $ 2.62万
• 依托单位: Osaka Bioscience Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670308/
• 关键词: Crk; PI3K; AKT; FAK; Src; SH2; SH3; Ras; v-CrK; P13K

v-Crk, an oncogene product of avian sarcoma virus CT10, transforms chicken embryo fibroblasts (CEF) efficiently. We have recently reported that phosphatidylinositide 3-kinase(PI3K)/AKT pathway is constitutively activated and play a critical role in the oncogenic transformation of CEF by v-Crk. In this study, we investigated how v-Crk activates PI3K/AKT pathway and revealed the involvement of focal adhesion kinase (FAK) and H-Ras in this process. v-Crk induces phosphorylation of Tyr 397 residue in FAK through the activation of src-family tyrosine kinase(s), in which SH2 domain of v-Crk is responsible, and promotes the binding of N-terminal SH2 domain of PI3K p85 regulatory subunit to this site. v-Crk fails to activate PI3K/AKT pathway in FAK null cells. In addition, we found that v-Crk stimulates the interaction of H-Ras with Ras binding domain in PI3K p110 catalytic subunit, and that the expression of H-Ras or its guanine nucleotide exchange factor mSOS, which binds to SH3 domain of v-Crk, enhances the v-Crk-induced activation of AKT, while a doninant negative mutant of H-Ras almost completely suppresses this activation. Our data demonstrated that v-Crk activates PI3K/AKT pathway by promoting both the interaction of p85 with tyrosine phosphorylated FAK and the interaction of p110 with H-Ras.

v-Crk是禽肉瘤病毒CT 10的癌基因产物，可有效转化鸡胚成纤维细胞（CEF）。我们最近报道，磷脂酰肌醇3-激酶（PI 3 K）/AKT通路被组成性激活，并在v-Crk致CEF的致癌转化中起关键作用。在本研究中，我们研究了v-Crk如何激活PI 3 K/AKT通路，并揭示了粘着斑激酶（FAK）和H-Ras参与了这一过程。v-Crk通过激活src家族酪氨酸激酶（其中v-Crk的SH 2结构域负责）诱导FAK中Tyr 397残基的磷酸化，并促进PI 3 K p85调节亚基的N端SH 2结构域与该位点的结合。v-Crk不能激活FAK无效细胞中的PI 3 K/AKT通路。此外，我们还发现v-Crk可刺激H-Ras与PI 3 K p110催化亚基中Ras结合域的相互作用，并且与v-Crk的SH 3结构域结合的H-Ras或其鸟嘌呤核苷酸交换因子mSOS的表达可增强v-Crk诱导的AKT活化，而H-Ras的一个失活突变体几乎完全抑制这种活化。我们的数据表明，v-Crk通过促进p85与酪氨酸磷酸化FAK的相互作用以及p110与H-Ras的相互作用来激活PI 3 K/AKT通路。

项目成果

Akagi, T., Murata, K., Shishido, T., Hanafusa, H.: "v-Crk activated the phosphoinositide 3-kinase/AKT pathway by utilizing focal adhesion kinase and H-Ras"Molecular and Cellular Biology. 22. 7015-7023 (2002)

Akagi, T.、Murata, K.、Shishido, T.、Hanafusa, H.：“v-Crk 通过利用粘着斑激酶和 H-Ras 激活磷酸肌醇 3-激酶/AKT 途径”分子和细胞生物学。

Shishido T, Akagi T, Chalmers A, Maeda M, Terada T, Georgescu MM, Hanafusa H.: "Crk family adaptor proteins trans-activate c-Abl kinase.."Genes Cells.. 6. 431-440 (2001)

Shishido T、Akagi T、Chalmers A、Maeda M、Terada T、Georgescu MM、Hanafusa H.：“Crk 家族接头蛋白反式激活 c-Abl 激酶..”Genes Cells.. 6. 431-440 (2001)

Akagi, T., Shishido, T., Murata, K., and Hanafusa, H.: "v-Crk activates the phosphoinositide 3-kinase/AKT pathway in transformation"PNAS. 97. 7290-7295 (2000)

Akagi, T.、Shishido, T.、Murata, K. 和 Hanafusa, H.：“v-Crk 在转化中激活磷酸肌醇 3-激酶/AKT 途径”PNAS。

Akagi T, Murata K, Shishido T,Hanafusa H.: "v-Crk Activates the Phosphoinositide 3-Kinase/AKT Pathway by Utilizing Focal Adhesion Kinase and H-Ras."Mol Cell Biol.. 22. 7015-7023 (2002)

Akagi T、Murata K、Shishido T、Hanafusa H.：“v-Crk 通过利用粘着斑激酶和 H-Ras 激活磷酸肌醇 3-激酶/AKT 途径。”Mol Cell Biol.. 22. 7015-7023 (2002)

Akagi T, Murata K, Shishido T, Hanafusa H: "v-Crk activated the phosphoinositide 3-kinase/AKT pathway by utilizing focal adhesion kinase and H-Ras"Molecular and Cellular Biology. 22. 7015-7023 (2002)

Akagi T、Murata K、Shishido T、Hanafusa H：“v-Crk 通过利用粘着斑激酶和 H-Ras 激活磷酸肌醇 3-激酶/AKT 途径”分子和细胞生物学。